Patent Publication Number: US-2003228597-A1

Title: Identification of genetic targets for modulation by oligonucleotides and generation of oligonucleotides for gene modulation

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
     [0001] The present application is a continuation-in-part of U.S. Ser. No. 09/295,463 filed Apr. 13, 1999, which is a continuation-in-part of U.S. Ser. No. 09/067,638 filed Apr. 28, 1998, which claims priority to provisional application Ser. No. 60/081,483 filed Apr. 13, 1998, each of which is incorporated herein by reference in its entirety. 
    
    
     
       FIELD OF THE INVENTION  
       [0002] The present invention relates generally to the generation and identification of synthetic compounds having defined physical, chemical or bioactive properties. More particularly, the present invention relates to the automated generation of oligonucleotide compounds targeted to a given nucleic acid sequence via computer-based, iterative robotic synthesis of synthetic oligonucleotide compounds and robotic or robot-assisted analysis of the activities of such compounds. Information gathered from assays of such compounds is used to identify nucleic acid sequences that are tractable to a variety of nucleotide sequence-based technologies, for example, gene function analysis and target validation.  
       BACKGROUND OF THE INVENTION  
       [0003] 1. Oligonucleotide Technology  
       [0004] Synthetic oligonucleotides of complementarity to targets are known to hybridize with particular, target nucleic acids in a sequence-specific manner. In one example, compounds complementary to the “sense” strand of nucleic acids that encode polypeptides, are referred to as “antisense oligonucleotides.” A subset of such compounds may be capable of modulating the expression of a target nucleic acid; such synthetic compounds are described herein as “active oligonucleotide compounds.” 
       [0005] Oligonucleotide compounds are commonly used in vitro as research reagents and diagnostic aids, and in vivo as therapeutic and bioactive agents. Oligonucleotide compounds can exert their effect by a variety of means. One such means takes advantage of an endogenous nuclease, such as RNase H in eukaryotes or RNase P in prokaryotes, to degrade the DNA/RNA hybrid formed between the oligonucleotide sequence and mRNA (Chiang et al.,  J. Biol. Chem.,  1991, 266, 18162; Forster et al.,  Science,  1990, 249, 783). Another means involves covalently linking of a synthetic moiety having nuclease activity to an oligonucleotide having an antisense sequence. This does not rely upon recruitment of an endogenous nuclease to modulate target activity. Synthetic moieties having nuclease activity include, but are not limited to, enzymatic RNAs, lanthanide ion complexes, and other reactive species. (Haseloff et al., Nature, 1988, 334, 585; Baker et al.,  J. Am. Chem. Soc.,  1997, 119, 8749).  
       [0006] Despite the advances made in utilizing antisense technology to date, it is still common to identify target sequences amenable to antisense technologies through an empirical approach (Szoka,  Nature Biotechnology,  1997, 15, 509). Accordingly, the need exists for systems and methods for efficiently and effectively identifying target nucleotide sequences that are suitable for antisense modulation. The present disclosure answers this need by providing systems and methods for automatically identifying such sequences via in silico, robotic or other automated means.  
       [0007] 2. Identification of Active Oligonucleotide Compounds  
       [0008] Traditionally, new chemical entities with useful properties are generated by (1) identifying a chemical compound (called a “lead compound”) with some desirable property or activity, (2) creating variants of the lead compound, and (3) evaluating the property and activity of such variant compounds. The process has been called “SAR,” i.e., structure activity relationship. Although “SAR” and its handmaiden, rational drug design, has been utilized with some degree of success, there are a number of limitations to these approaches to lead compound generation, particularly as it pertains to the discovery of bioactive oligonucleotide compounds. In attempting to use SAR with oligonucleotides, it has been recognized that RNA structure can inhibit duplex formation with antisense compounds, so much so that “moving” the target nucleotide sequence even a few bases can drastically decrease the activity of such compounds (Lima et al.,  Biochemistry,  1992, 31, 12055).  
       [0009] Heretofore, the preferred method of searching for lead antisense compounds has been the manual synthesis and analysis of such compounds. Consequently, a fundamental limitation of the conventional approach is its dependence upon the availability, number and cost of antisense compounds produced by manual, or at best semi-automated, means. Moreover, the assaying of such compounds has traditionally been performed by tedious manual techniques. Thus, the traditional approach to generating active antisense compounds is limited by the relatively high cost and long time required to synthesize and screen a relatively small number of candidate antisense compounds.  
       [0010] Accordingly, the need exists for systems and methods for efficiently and effectively generating new active antisense and other oligonucleotide compounds targeted to specific nucleic acid sequences. The present disclosure answers this need by providing systems and methods for automatically generating and screening active antisense compounds via robotic and other automated means.  
       [0011] 3. Gene Function Analysis  
       [0012] Efforts such as the Human Genome Project are making an enormous amount of nucleotide sequence information available in a variety of forms, e.g., genomic sequences, cDNAs, expressed sequence tags (ESTs) and the like. This explosion of information has led one commentator to state that “genome scientists are producing more genes than they can put a function to” (Kahn,  Science,  1995, 270, 369). Although some approaches to this problem have been suggested, no solution has yet emerged. For example, methods of looking at gene expression in different disease states or stages of development only provide, at best, an association between a gene and a disease or stage of development (Nowak,  Science,  1995, 270, 368). Another approach, looking at the proteins encoded by genes, is developing but “this approach is more complex and big obstacles remain” (Kahn,  Science,  1995, 270, 369). Furthermore, neither of these approaches allows one to directly utilize nucleotide sequence information to perform gene function analysis.  
       [0013] In contrast, antisense technology does allow for the direct utilization of nucleotide sequence information for gene function analysis. Once a target nucleic acid sequence has been selected, antisense sequences hybridizable to the sequence can be generated using techniques known in the art. Typically, a large number of candidate antisense oligonucleotides (ASOs) are synthesized having sequences that are more-or-less randomly spaced across the length of the target nucleic acid sequence (e.g., a “gene walk”) and their ability to modulate the expression of the target nucleic acid is assayed. Cells or animals can then be treated with one or more active antisense oligonucleotides, and the resulting effects determined in order to determine the function(s) of the target gene. Although the practicality and value of this empirical approach to determining gene function has been acknowledged in the art, it has also been stated that this approach “is beyond the means of most laboratories and is not feasible when a new gene sequence is identified, but whose function and therapeutic potential are unknown” (Szoka,  Nature Biotechnology,  1997, 15, 509).  
       [0014] Accordingly, the need exists for systems and methods for efficiently and effectively determining the function of a gene that is uncharacterized except that its nucleotide sequence, or a portion thereof, is known. The present disclosure answers this need by providing systems and methods for automatically generating active antisense compounds to a target nucleotide sequence via robotic means. Such active antisense compounds are contacted with cells, cell-free extracts, tissues or animals capable of expressing the gene of interest and subsequent biochemical or biological parameters are measured. The results are compared to those obtained from a control cell culture, cell-free extract, tissue or animal which has not been contacted with an active antisense compound in order to determine the function of the gene of interest.  
       [0015] 4. Target Validation  
       [0016] Determining the nucleotide sequence of a gene is no longer an end unto itself; rather, it is “merely a means to an end. The critical next step is to validate the gene and its [gene] product as a potential drug target” (Glasser,  Genetic Engineering News,  1997, 17, 1). This process, i.e., confirming that modulation of a gene that is suspected of being involved in a disease or disorder actually results in an effect that is consistent with a causal relationship between the gene and the disease or disorder, is known as target validation.  
       [0017] Efforts such as the Human Genome Project are yielding a vast number of complete or partial nucleotide sequences, many of which might correspond to or encode targets useful for new drug discovery efforts. The challenge represented by this plethora of information is how to use such nucleotide sequences to identify and rank valid targets for drug discovery. Antisense technology provides one means by which this might be accomplished; however, the many manual, labor-intensive and costly steps involved in traditional methods of developing active antisense compounds has limited their use in target validation (Szoka,  Nature Biotechnology,  1997, 15, 509). Nevertheless, the great target specificity that is characteristic of antisense compounds makes them ideal choices for target validation, especially when the functional roles of proteins that are highly related are being investigated (Albert et al.,  Trends in Pharm. Sci.,  1994, 15, 250).  
       [0018] Accordingly, the need exists for systems and methods for developing compounds efficiently and effectively that modulate a gene, wherein such compounds can be directly developed from nucleotide sequence information. Such compounds are needed to confirm that modulation of a gene that is thought to be involved in a disease or disorder will in fact cause an in vitro or in vivo effect indicative of the origin, development, spread or growth of the disease or disorder.  
       [0019] The present disclosure answers this need by providing systems and methods for automatically generating active oligonucleotide and other compounds, especially antisense compounds, to a target nucleotide sequence via robotic or other automated means. Such active compounds are contacted with a cell culture, cell-free extract, tissue or animal capable of expressing the gene of interest, and subsequent biochemical or biological parameters indicative of the potential gene product function are measured. These results are compared to those obtained with a control cell system, cell-free extract, tissue or animal which has not been contacted with an active antisense compound in order to determine whether or not modulation of the gene of interest affects a specific cellular function. The resulting active antisense compounds may be used as positive controls when other, non antisense-based agents directed to the same target nucleic acid, or to its gene product, are screened.  
       [0020] It should be noted that embodiments of the invention drawn to gene function analysis and target validation have parameters that are shared with other embodiments of the invention, but also have unique parameters. For example, antisense drug discovery naturally requires that the toxicity of the antisense compounds be manageable, whereas, for gene function analysis or target validation, overt toxicity resulting from the antisense compounds is acceptable unless it interferes with the assay being used to evaluate the effects of treatment with such compounds.  
       [0021] U.S. Pat. No. 5,563,036 to Peterson et al. describes systems and methods of screening for compounds that inhibit the binding of a transcription factor to a nucleic acid. In a preferred embodiment, an assay portion of the process is stated to be performed by a computer controlled robot.  
       [0022] U.S. Pat. No. 5,708,158 to Hoey describes systems and methods for identifying pharmacological agents stated to be useful for diagnosing or treating a disease associated with a gene the expression of which is modulated by a human nuclear factor of activated T cells. The methods are stated to be particularly suited to high-thoughput screening wherein one or more steps of the process are performed by a computer controlled robot.  
       [0023] U.S. Pat. Nos. 5,693,463 and 5,716,780 to Edwards et al. describe systems and methods for identifying non-oligonucleotide molecules that specifically bind to a DNA molecule based on their ability to compete with a DNA-binding protein that recognizes the DNA molecule.  
       [0024] U.S. Pat. Nos. 5,463,564 and 5,684,711 to Agrafiotis et al. describe computer based iterative processes for generating chemical entities with defined physical, chemical and/or bioactive properties.  
       [0025] 5. Compounds of the Invention  
       [0026] According to the present invention, compounds include antisense oligomeric compounds, antisense oligonucleotides, ribozymes, external guide sequence (EGS) oligonucleotides, alternate splicers, primers, probes, and other oligomeric compounds which hybridize to at least a portion of the target nucleic acid. As such, these compounds may be introduced in the form of single-stranded, double-stranded, circular or hairpin oligomeric compounds and may contain structural elements such as internal or terminal bulges or loops. Once introduced to a system, the compounds of the invention may elicit the action of one or more enzymes or structural proteins to effect modification of the target nucleic acid.  
       [0027] One non-limiting example of such an enzyme is RNAse H, a cellular endonuclease which cleaves the RNA strand of an RNA:DNA duplex. It is known in the art that single-stranded antisense compounds which are “DNA-like” elicit RNAse H. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of oligonucleotide-mediated inhibition of gene expression. Similar roles have been postulated for other ribonucleases such as those in the RNase III and ribonuclease L family of enzymes.  
       [0028] While the preferred form of antisense compound is a single-stranded antisense oligonucleotide, in many species the introduction of double-stranded structures, such as double-stranded RNA (dsRNA) molecules, has been shown to induce potent and specific antisense-mediated reduction of the function of a gene or its associated gene products. This phenomenon occurs in both plants and animals and is believed to have an evolutionary connection to viral defense and transposon silencing.  
       [0029] The first evidence that dsRNA could lead to gene silencing in animals came in 1995 from work in the nematode, Caenorhabditis elegans (Guo and Kempheus, Cell, 1995, 81, 611-620). Montgomery et al. have shown that the primary interference effects of dsRNA are posttranscriptional (Montgomery et al., Proc. Natl. Acad. Sci. USA, 1998, 95, 15502-15507). The posttranscriptional antisense mechanism defined in Caenorhabditis elegans resulting from exposure to double-stranded RNA (dsRNA) has since been designated RNA interference (RNAi). This term has been generalized to mean antisense-mediated gene silencing involving the introduction of dsRNA leading to the sequence-specific reduction of endogenous targeted mRNA levels (Fire et al., Nature, 1998, 391, 806-811). Recently, it has been shown that it is, in fact, the single-stranded RNA oligomers of antisense polarity of the dsRNAs which are the potent inducers of RNAi (Tijsterman et al., Science, 2002, 295, 694-697).  
       [0030] In the context of this invention, the term “oligomeric compound” refers to a polymer or oligomer comprising a plurality of monomeric units. In the context of this invention, the term “oligonucleotide” refers to an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or mimetics, chimeras, analogs and homologs thereof. This term includes oligonucleotides composed of naturally occurring nucleobases, sugars and covalent internucleoside (backbone) linkages as well as oligonucleotides having non-naturally occurring portions which function similarly. Such modified or substituted oligonucleotides are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for a target nucleic acid and increased stability in the presence of nucleases.  
       [0031] While oligonucleotides are a preferred form of the compounds of this invention, the present invention comprehends other families of compounds as well, including but not limited to oligonucleotide analogs and mimetics such as those described herein.  
       [0032] The compounds in accordance with this invention preferably comprise from about 8 to about 80 nucleobases (i.e. from about 8 to about 80 linked nucleosides). One of ordinary skill in the art will appreciate that the invention embodies compounds of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 nucleobases in length.  
       [0033] In one preferred embodiment, the compounds of the invention are 12 to 50 nucleobases in length. One having ordinary skill in the art will appreciate that this embodies compounds of 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleobases in length.  
       [0034] In another preferred embodiment, the compounds of the invention are 15 to 30 nucleobases in length. One having ordinary skill in the art will appreciate that this embodies compounds of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length.  
       [0035] In another preferred embodiment, the compounds of the invention are 20 to 25 nucleobases in length. One having ordinary skill in the art will appreciate that this embodies compounds of 20, 21, 22, 23, 24 or 25 nucleobases in length.  
       [0036] Particularly preferred compounds are oligonucleotides from about 12 to about 50 nucleobases, even more preferably those comprising from about 15 to about 30 nucleobases.  
       [0037] Antisense compounds 8-80 nucleobases in length comprising a stretch of at least eight (8) consecutive nucleobases selected from within the illustrative antisense compounds are considered to be suitable antisense compounds as well.  
       [0038] Exemplary preferred antisense compounds include oligonucleotide sequences that comprise at least the 8 consecutive nucleobases from the 5′-terminus of one of the illustrative preferred antisense compounds (the remaining nucleobases being a consecutive stretch of the same oligonucleotide beginning immediately upstream of the 5′-terminus of the antisense compound which is specifically hybridizable to the target nucleic acid and continuing until the oligonucleotide contains about 8 to about 80 nucleobases). Similarly preferred antisense compounds are represented by oligonucleotide sequences that comprise at least the 8 consecutive nucleobases from the 3′-terminus of one of the illustrative preferred antisense compounds (the remaining nucleobases being a consecutive stretch of the same oligonucleotide beginning immediately downstream of the 3′-terminus of the antisense compound which is specifically hybridizable to the target nucleic acid and continuing until the oligonucleotide contains about 8 to about 80 nucleobases). One having skill in the art armed with the preferred antisense compounds illustrated herein will be able, without undue experimentation, to identify further preferred antisense compounds.  
       SUMMARY OF THE INVENTION  
       [0039] The present invention is directed to methods of effecting gene function analysis and target validation by generating in silico a library of nucleobase sequences targeted to the gene and robotically assaying a plurality of synthetic compounds having at least some of the nucleobase sequences to identify target modulators. These modulators are then assayed for effects on biological function to effect gene function analysis and for effects on diseases or disorders to effect target validation.  
       [0040] Further features and advantages of the present invention, as well as the structure and operation of various embodiments of the present invention, are described in detail below with reference to the accompanying drawings. In the drawings, like reference numbers indicate identical or functionally similar elements. 
     
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
     [0041] The present invention will be described with reference to the accompanying drawings, wherein:  
     [0042]FIGS. 1 and 2 are a flow diagram of one method according to the present invention depicting the overall flow of data and materials among various elements of the invention.  
     [0043]FIG. 3 is a flow diagram depicting the flow of data and materials among elements of step  200  of FIG. 1.  
     [0044]FIGS. 4 and 5 are a flow diagram depicting the flow of data and materials among elements of step  300  of FIG. 1.  
     [0045]FIG. 6 is a flow diagram depicting the flow of data and materials among elements of step  306  of FIG. 4.  
     [0046]FIG. 7 is another flow diagram depicting the flow of data and materials among elements of step  306  of FIG. 4.  
     [0047]FIG. 8 is a another flow diagram depicting the flow of data and materials among elements of step  306  of FIG. 4.  
     [0048]FIG. 9 is a flow diagram depicting the flow of data and materials among elements of step  350  of FIG. 5.  
     [0049]FIGS. 10 and 11 are flow diagrams depicting a logical analysis of data and materials among elements of step  400  of FIG. 1.  
     [0050]FIG. 12 is a flow diagram depicting the flow of data and materials among the elements of step  400  of FIG. 1.  
     [0051]FIGS. 13 and 14 are flow diagrams depicting the flow of data and materials among elements of step  500  of FIG. 1.  
     [0052]FIG. 15 is a flow diagram depicting the flow of data and materials among elements of step  600  of FIG. 1.  
     [0053]FIG. 16 is a flow diagram depicting the flow of data and materials among elements of step  700  of FIG. 1.  
     [0054]FIG. 17 is a flow diagram depicting the flow of data and materials among the elements of step  1100  of FIG. 2.  
     [0055]FIG. 18 is a block diagram showing the interconnecting of certain devices utilized in conjunction with a preferred method of the invention;  
     [0056]FIG. 19 is a flow diagram showing a representation of data storage in a relational database utilized in conjunction with one method of the invention;  
     [0057]FIG. 20 is a flow diagram depicting the flow of data and materials in effecting a preferred embodiment of the invention as set forth in Example 16;  
     [0058]FIG. 21 is a flow diagram depicting the flow of data and materials in effecting a preferred embodiment of the invention as set forth in Example 17;  
     [0059]FIG. 22 is a flow diagram depicting the flow of data and materials in effecting a preferred embodiment of the invention as set forth in Example 2;  
     [0060]FIG. 23 is a pictorial elevation view of a preferred apparatus used to robotically synthesize oligonucleotides; and  
     [0061]FIG. 24 is a pictorial plan view of an apparatus used to robotically synthesize oligonucleotides. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
     [0062] Certain preferred methods of this invention are now described with reference to the flow diagram of FIGS. 1 and 2.  
     [0063] 1. Target Nucleic Acid Selection.  
     [0064] The target selection process, process step  100 , provides a target nucleotide sequence that is used to help guide subsequent steps of the process. It is generally desired to modulate the expression of the target nucleic acid for any of a variety of purposes, such as, e.g., drug discovery, target validation and/or gene function analysis.  
     [0065] One of the primary objectives of the target selection process, step  100 , is to identify molecular targets that represent significant therapeutic opportunities, to provide new and efficacious means of drug discovery and to determine the function of genes that are uncharacterized except for nucleotide sequence. To meet these objectives, genes are classified based upon specific sets of selection criteria.  
     [0066] One such set of selection criteria concerns the quantity and quality of target nucleotide sequence. There must be sufficient target nucleic acid sequence information available for oligonucleotide design. Moreover, such information must be of sufficient quality to give rise to an acceptable level of confidence in the data to perform the methods described herein. Thus, the data must not contain too many missing or incorrect base entries. In the case of a target sequence that encodes a polypeptide, such errors can often be detected by virtually translating all three reading frames of the sense strand of the target sequence and confirming the presence of a continuous polypeptide sequence having predictable attributes, e.g., encoding a polypeptide of known size, or encoding a polypeptide that is about the same length as a homologous protein. In any event, only a very high frequency of sequence errors will frustrate the methods of the invention; most oligonucleotides to the target sequence will avoid-such errors unless such errors occur frequently throughout the entire target sequence.  
     [0067] Another preferred criterion is that appropriate culturable cell lines or other source of reproducible genetic expression should be available. Such cell lines express, or can be induced to express, the gene comprising the target nucleic acid sequence. The oligonucleotide compounds generated by the process of the invention are assayed using such cell lines and, if such assaying is performed robotically, the cell line is preferably tractable to robotic manipulation such as by growth in 96 well plates. Those skilled in the art will recognize that if an appropriate cell line does not exist, it will nevertheless be possible to construct an appropriate cell line. For example, a cell line can be transfected with an expression vector comprising the target gene in order to generate an appropriate cell line for assay purposes.  
     [0068] For gene function analysis, it is possible to operate upon a genetic system having a lack of information regarding, or incomplete characterization of, the biological function(s) of the target nucleic acid or its gene product(s). This is a powerful agent of the invention. A target nucleic acid for gene function analysis might be absolutely uncharacterized, or might be thought to have a function based on minimal data or homology to another gene. By application of the process of the invention to such a target, active compounds that modulate the expression of the gene can be developed and applied to cells. The resulting cellular, biochemical or molecular biological responses are observed, and this information is used by those skilled in the art to elucidate the function of the target gene.  
     [0069] For target validation and drug discovery, another selection criterion is disease association. Candidate target genes are placed into one of several broad categories of known or deduced disease association. Level 1 Targets are target nucleic acids for which there is a strong correlation with disease. This correlation can come from multiple scientific disciplines including, but not limited to, epidemiology, wherein frequencies of gene abnormalities are associated with disease incidence; molecular biology, wherein gene expression and function are associated with cellular events correlated with a disease; and biochemistry, wherein the in vitro activities of a gene product are associated with disease parameters. Because there is a strong therapeutic rationale for focusing on Level 1 Targets, these targets are most preferred for drug discovery and/or target validation.  
     [0070] Level 2 Targets are nucleic acid targets for which the combined epidemiological, molecular biological, and/or biochemical correlation with disease is not so clear as for Level 1. Level 3 Targets are targets for which there is little or no data to directly link the target with a disease process, but there is indirect evidence for such a link, i.e., homology with a Level 1 or Level 2 target nucleic acid sequence or with the gene product thereof. In order not to prejudice the target selection process, and to ensure that the maximum number of nucleic acids actually involved in the causation, potentiation, aggravation, spread, continuance or after-effects of disease states are investigated, it is preferred to examine a balanced mix of Level 1, 2 and 3 target nucleic acids.  
     [0071] In order to carry out drug discovery, experimental systems and reagents shall be available in order for one to evaluate the therapeutic potential of active compounds generated by the process of the invention. Such systems may be operable in vitro (e.g., in vitro models of cell:cell association) or in vivo (e.g., animal models of disease states). It is also desirable, but not obligatory, to have available animal model systems which can be used to evaluate drug pharmacology.  
     [0072] Candidate targets nucleic acids can also classified by biological processes. For example, programmed cell death (“apoptosis”) has recently emerged as an important biological process that is perturbed in a wide variety of diseases. Accordingly, nucleic acids that encode factors that play a role in the apoptotic process are identified as candidate targets. Similarly, potential target nucleic acids can be classified as being involved in inflammation, autoimmune disorders, cancer, or other pathological or dysfunctional processes.  
     [0073] Moreover, genes can often be grouped into families based on sequence homology and biological function. Individual family members can act redundantly, or can provide specificity through diversity of interactions with downstream effectors, or through expression being restricted to specific cell types. When one member of a gene family is associated with a disease process then the rationale for targeting other members of the same family is reasonably strong. Therefore, members of such gene families are preferred target nucleic acids to which the methods and systems of the invention may be applied. Indeed, the potent specificity of antisense compounds for different gene family members makes the invention particularly suited for such targets (Albert et al.,  Trends Pharm. Sci.,  1994, 15, 250). Those skilled in the art will recognize that a partial or complete nucleotide sequence of such family members can be obtained using the polymerase chain reaction (PCR) and “universal” primers, i.e., primers designed to be common to all members of a given gene family.  
     [0074] PCR products generated from universal primers can be cloned and sequenced or directly sequenced using techniques known in the art. Thus, although nucleotide sequences from cloned DNAs, or from complementary DNAs (cDNAs) derived from mRNAs, may be used in the process of the invention, there is no requirement that the target nucleotide sequence be isolated from a cloned nucleic acid. Any nucleotide sequence, no matter how determined, of any nucleic acid, isolated or prepared in any fashion, may be used as a target nucleic acid in the process of the invention.  
     [0075] Furthermore, although polypeptide-encoding nucleic acids provide the target nucleotide sequences in one embodiment of the invention, other nucleic acids may be targeted as well. Thus, for example, the nucleotide sequences of structural or enzymatic RNAs may be utilized for drug discovery and/or target validation when such RNAs are associated with a disease state, or for gene function analysis when their biological role is not known.  
     [0076] 2. Assembly of Target Nucleotide Sequence.  
     [0077]FIG. 3 is a block diagram detailing the steps of the target nucleotide sequence assembly process, process step  200  in acccordance with one embodiment of the invention. The oligonucleotide design process, process step  300 , is facilitated by the availability of accurate target sequence information. Because of limitations of automated genome sequencing technology, gene sequences are often accumulated in fragments. Further, because individual genes are often being sequenced by independent laboratories using different sequencing strategies, sequence information corresponding to different fragments is often deposited in different databases. The target nucleic acid assembly process take advantage of computerized homology search algorithms and sequence fragment assembly algorithms to search available databases for related sequence information and incorporate available sequence information into the best possible representation of the target nucleic acid molecule, for example a RNA transcript. This representation is then used to design oligonucleotides, process step  300 , which can be tested for biological activity in process step  700 .  
     [0078] In the case of genes directing the synthesis of multiple transcripts, i.e. by alternative splicing, each distinct transcript is a unique target nucleic acid for purposes of step  300 . In one embodiment of the invention, if active compounds specific for a given transcript isoform are desired, the target nucleotide sequence is limited to those sequences that are unique to that transcript isoform. In another embodiment of the invention, if it is desired to modulate two or more transcript isoforms in concert, the target nucleotide sequence is limited to sequences that are shared between the two or more transcripts.  
     [0079] In the case of a polypeptide-encoding nucleic acid, it is generally preferred that full-length cDNA be used in the oligonucleotide design process step  300  (with full-length cDNA being defined as reading from the 5′ cap to the poly A tail). Although full-length cDNA is preferred, it is possible to design oligonucleotides using partial sequence information. Therefore it is not necessary for the assembly process to generate a complete cDNA sequence. Further in some cases it may be desirable to design oligonucleotides targeting introns. In this case the process can be used to identify individual introns at process step  220 .  
     [0080] The process can be initiated by entering initial sequence information on a selected molecular target at process step  205 . In the case of a polypeptide-encoding nucleic acid, the full-length cDNA sequence is generally preferred for use in oligonucleotide design strategies at process step  300 . The first step is to determine if the initial sequence information represents the full-length cDNA, decision step  210 . In the case where the full-length cDNA sequence is available the process advances directly to the oligonucleotide design step  300 . When the full-length cDNA sequence is not available, databases are searched at process step  212  for additional sequence information.  
     [0081] The algorithm preferably used in process steps  212  and  230  is BLAST (Altschul, et al.,  J. Mol. Biol.,  1990, 215, 403), or “Gapped BLAST” (Altschul et al.,  Nucl. Acids Res.,  1997, 25, 3389). These are database search tools based on sequence homology used to identify related sequences in a sequence database. The BLAST search parameters are set to only identify closely related sequences. Some preferred databases searched by BLAST are a combination of public domain and proprietary databases. The databases, their contents, and sources are listed in Table 1.  
               TABLE 1                          Database Sources of Target Sequences                         Database   Contents   Source               NR   All non-redundant   National Center for           GenBank, EMBL, DDBJ   Biotechnology Information at the           and PDB sequences   National Institutes of Health       Month   All new or revised   National Center for           GenBank, EMBL, DDBJ   Biotechnology Information at the           and PDB sequences   National Institutes of Health           released in the           last 30 days       Dbest   Non-redundant   National Center for           database of GenBank,   Biotechnology Information at the           EMBL, DDBJ and   National Institutes of Health           EST divisions       Dbsts   Non-redundant   National Center for           database of GenBank,   Biotechnology Information at the           EMBL, DDBJ and   National Institutes of Health           STS divisions       Htgs   High throughput   National Center for           genomic sequences   Biotechnology Information at the               National Institutes of Health                  
 
     [0082] When genomic sequence information is available at decision step  215 , introns are removed and exons are assembled into continuous sequence representing the cDNA sequence in process step  220 . Exon assembly occurs using the Phragment Assembly  P rogram “Phrap” (Copyright University of Washington Genome Center, Seattle, Wash.). The Phrap algorithm analyzes sets of overlapping sequences and assembles them into one continuous sequence referred to as a “contig.” The resulting contig is preferably used to search databases for additional sequence information at process step  230 . When genomic information is not available, the results of process step  212  are analyzed for individual exons at decision step  225 . Exons are frequently recorded individually in databases. If multiple complete exons are identified, they are prferably assembled into a contig using Phrap at process step  250 . If multiple complete exons are not identified at decision step  225 , then sequences can be analyzed for partial sequence information in decision step  228 . ESTs identified in the database dbEST are examples of such partial sequence information. If additional partial information is not found, then the process is advanced to process step  230  at decision step  228 . If partial sequence information is found in process  212  then that information is advanced to process step  230  via decision step  228 .  
     [0083] Process step  230 , decision step  240 , decision step  260  and process step  250  define a loop designed to extend iteratively the amount of sequence information available for targeting. At the end of each iteration of this loop, the results are analyzed in decision steps  240  and  260 . If no new information is found then the process advances at decision step  240  to process step  300 . If there is an unexpectedly large amount of sequence information identified, suggesting that the process moved outside the boundary of the gene into repetitive genomic sequence, then the process is preferably cycled back one iteration and that sequence is advanced at decision step  240  to process step  300 . If a small amount of new sequence information is identified, then the loop is iterated such as by taking the 100 most 5-prime (5′) and 100 most 3-prime (3′) bases and interating them through the BLAST homology search at process step  230 . New sequence information is added to the existing contig at process step  250 .  
     [0084] 3. In Silico Generation of a Set of Nucleobase Sequences and Virtual Oligonucleotides.  
     [0085] For the following steps  300  and  400 , they may be performed in the order described below, i.e., step  300  before step  400 , or, in an alternative embodiment of the invention, step  400  before step  300 . In this alternate embodiment, each oligonucleotide chemistry is first assigned to each oligonucleotide sequence. Then, each combination of oligonucleotide chemistry and sequence is evaluated according to the parameters of step  300 . This embodiment has the desirable feature of taking into account the effect of alternative oligonucleotide chemistries on such parameters. For example, substitution of 5-methyl cytosine (5MeC or m5c) for cytosine in an antisense compound may enhance the stability of a duplex formed between that compound and its target nucleic acid. Other oligonucleotide chemistries that enhance oligonucleotide:[target nucleic acid] duplexes are known in the art (see for example, Freier et al.,  Nucleic Acids Research,  1997, 25, 4429). As will be appreciated by those skilled in the art, different oligonucleotide chemistries may be preferred for different target nucleic acids. That is, the optimal oligonucleotide chemistry for binding to a target DNA might be suboptimal for binding to a target RNA having the same nucleotide sequence.  
     [0086] In effecting the process of the invention in the order step  300  before step  400  as seen in FIG. 1, from a target nucleic acid sequence assembled at step  200 , a list of oligonucleotide sequences is generated as represented in the flowchart shown in FIGS. 4 and 5. In step  302 , the desired oligonucleotide length is chosen. In a preferred embodiment, oligonucleotide length is between from about 8 to about 30, more preferably from about 12 to about 25, nucleotides. In step  304 , all possible oligonucleotide sequences of the desired length capable of hybridizing to the target sequence obtained in step  200  are generated. In this step, a series of oligonucleotide sequences are generated, simply by determining the most 5′ oligonucleotide possible and “walking” the target sequence in increments of one base until the 3′ most oligonucleotide possible is reached.  
     [0087] In step  305 , a virtual oligonucleotide chemistry is applied to the nucleobase sequences of step  304  in order to yield a set of virtual oligonucleotides that can be evaluated in silico. Default virtual oligonucleotide chemistries include those that are well-characterized in terms of their physical and chemical properties, e.g., 2′-deoxyribonucleic acid having naturally occurring bases (A, T, C and G), unmodified sugar residues and a phosphodiester backbone.  
     [0088] 4. In Silico Evaluation of Thermodynamic Properties of Virtual Oligonucleotides.  
     [0089] In step  306 , a series of thermodynamic, sequence, and homology scores are preferably calculated for each virtual oligonucleotide obtained from step  305 . Thermodynamic properties are calculated as represented in FIG. 6. In step  308 , the desired thermodynamic properties are selected. As many or as few as desired can be selected; optionally, none will be selected. The desired properties will typically include step  309 , calculation of the free energy of the target structure. If the oligonucleotide is a DNA molecule, then steps  310 ,  312 , and  314  are performed. If the oligonucleotide is an RNA molecule, then steps  311 ,  313  and  315  are performed. In both cases, these steps correspond to calculation of the free energy of intramolecular oligonucleotide interactions, intermolecular interactions and duplex formation. In addition, a free energy of oligonucleotide-target binding is preferably calculated at step  316 .  
     [0090] Other thermodynamic and kinetic properties may be calculated for oligonucleotides as represented at step  317 . Such other thermodynamic and kinetic properties may include melting temperatures, association rates, dissociation rates, or any other physical property that may be predictive of oligonucleotide activity.  
     [0091] The free energy of the target structure is defined as the free energy needed to disrupt any secondary structure in the target binding site of the targeted nucleic acid. This region includes any intra-target nucleotide base pairs that need to be disrupted in order for an oligonucleotide to bind to its complementary sequence. The effect of this localized disruption of secondary structure is to provide accessibility by the oligonucleotide. Such structures will include double helices, terminal unpaired and mismatched nucleotides, loops, including hairpin loops, bulge loops, internal loops and multibranch loops (Serra et al.,  Methods in Enzymology,  1995, 259, 242).  
     [0092] The intermolecular free energies refer to inherent energy due to the most stable structure formed by two oligonucleotides; such structures include dimer formation. Intermolecular free energies should also be taken into account when, for example, two or more oligonucleotides, of different sequence are to be administered to the same cell in an assay.  
     [0093] The intramolecular free energies refer to the energy needed to disrupt the most stable secondary structure within a single oligonucleotide. Such structures include, for example, hairpin loops, bulges and internal loops. The degree of intramolecular base pairing is indicative of the energy needed to disrupt such base pairing.  
     [0094] The free energy of duplex formation is the free energy of denatured oligonucleotide binding to its denatured target sequence. The oligonucleotide-target binding is the total binding involved, and includes the energies involved in opening up intra- and inter-molecular oligonucleotide structures, opening up target structure, and duplex formation.  
     [0095] The most stable RNA structure is predicted based on nearest neighbor analysis (Xia, T., et al.,  Biochemistry,  1998, 37, 14719-14735; Serra et al.,  Methods in Enzymology,  1995, 259, 242). This analysis is based on the assumption that stability of a given base pair is determined by the adjacent base pairs. For each possible nearest neighbor combination, thermodynamic properties have been determined and are provided. For double helical regions, two additional factors need to be considered, an entropy change required to initiate a helix and a entropy change associated with self-complementary strands only. Thus, the free energy of a duplex can be calculated using the equation: where:  
     [0096] ΔG is the free energy of duplex formation,  
     [0097] ΔH is the enthalpy change for each nearest neighbor,  
     [0098] ΔS is the entropy change for each nearest neighbor, and T is temperature.  
     [0099] The ΔH and ΔS for each possible nearest neighbor combination have been experimentally determined. These letter values are often available in published tables. For terminal unpaired and mismatched nucleotides, enthalpy and entropy measurements for each possible nucleotide combination are also available in published tables. Such results are added directly to values determined for duplex formation. For loops, while the available data is not as complete or accurate as for base pairing, one known model determines the free energy of loop formation as the sum of free energy based on loop size, the closing base pair, the interactions between the first mismatch of the loop with the closing base pair, and additional factors including being closed by AU or UA or a first mismatch of GA or UU. Such equations may also be used for oligoribonucleotide-target RNA interactions.  
     [0100] The stability of DNA duplexes is used in the case of intra- or intermolecular oligodeoxyribonucleotide interactions. DNA duplex stability is calculated using similar equations as RNA stability, except experimentally determined values differ between nearest neighbors in DNA and RNA and helix initiation tends to be more favorable in DNA than in RNA (SantaLucia et al.,  Biochemistry,  1996, 35, 3555).  
     [0101] Additional thermodynamic parameters are used in the case of RNA/DNA hybrid duplexes. This would be the case for an RNA target and oligodeoxynucleotide. Such parameters were determined by Sugimoto et al. ( Biochemistry,  1995, 34, 11211). In addition to values for nearest neighbors, differences were seen for values for enthalpy of helix initiation.  
     [0102] 5. In Silico Evaluation of Target Accessibility  
     [0103] Target accessibility is believed to be an important consideration in selecting oligonucleotides. Such a target site will possess minimal secondary structure and thus, will require minimal energy to disrupt such structure. In addition, secondary structure in oligonucleotides, whether inter- or intra-molecular, is undesirable due to the energy required to disrupt such structures. Oligonucleotide-target binding is dependent on both these factors. It is desirable to minimize the contributions of secondary structure based on these factors. The other contribution to oligonucleotide-target binding is binding affinity. Favorable binding affinities based on tighter base pairing at the target site is desirable.  
     [0104] Following the calculation of thermodynamic properties ending at step  317 , the desired sequence properties to be scored are selected at step  324 . As many or as few as desired can be selected; optionally, none will be selected. These properties include the number of strings of four guanosine residues in a row at step  325  or three guanosine in a row at step  326 , the length of the longest string of adenosines at step  327 , cytidines at step  328  or uridines or thymidines at step  329 , the length of the longest string of purines at step  330  or pyrimidine at step  331 , the percent composition of adenosine at step  332 , cytidine at step  333 , guanosine at step  334  or uridines or thymidines at step  335 , the percent composition of purines at step  336  or pyrimidines at step  337 , the number of CG dinucleotide repeats at step  338 , CA dinucleotide repeats at step  339  or UA or TA dinucleotide repeats at step  340 . In addition, other sequence properties may be used as found to be relevant and predictive of antisense efficacy, as represented at step  341 .  
     [0105] These sequence properties may be important in predicting oligonucleotide activity, or lack thereof. For example, U.S. Pat. No. 5,523,389 discloses oligonucleotides containing stretches of three or four guanosine residues in a row. Oligonucleotides having such sequences may act in a sequence-independent manner. For an antisense approach, such a mechanism is not usually desired. In addition, high numbers of dinucleotide repeats may be indicative of low complexity regions which may be present in large numbers of unrelated genes. Unequal base composition, for example, 90% adenosine, can also give non-specific effects. From a practical standpoint, it may be desirable to remove oligonucleotides that possess long stretches of other nucleotides due to synthesis considerations. Other sequences properties, either listed above or later found to be of predictive value may be used to select oligonucleotide sequences.  
     [0106] Following step  341 , the homology scores to be calculated are selected in step  342 . Homology to nucleic acids encoding protein isoforms of the target, as represented at step  343 , may be desired. For example, oligonucleotides specific for an isoform of protein kinase C can be selected. Also, oligonucleotides can be selected to target multiple isoforms of such genes. Homology to analogous target sequences, as represented at step  344 , may also be desired. For example, an oligonucleotide can be selected to a region common to both humans and mice to facilitate testing of the oligonucleotide in both species. Homology to splice variants of the target nucleic acid, as represented at step  345 , may be desired. In addition, it may be desirable to determine homology to other sequence variants as necessary, as represented in step  346 .  
     [0107] Following step  346 , from which scores were obtained in each selected parameter, a desired range is selected to select the most promising oligonucleotides, as represented at step  347 . Typically, only several parameters will be used to select oligonucleotide sequences. As structure prediction improves, additional parameters may be used. Once the desired score ranges are chosen, a list of all oligonucleotides having parameters falling within those ranges will be generated, as represented at step  348 .  
     [0108] 6. Targeting Oligonucleotides to Functional Regions of a Nucleic Acid.  
     [0109] It may be desirable to target oligonucleotide sequences to specific functional regions of the target nucleic acid. A decision is made whether to target such regions, as represented in decision step  349 . If it is desired to target functional regions then process step  350  occurs as seen in greater detail in FIG. 9. If it is not desired then the process proceeds to step  375 .  
     [0110] In step  350 , as seen in FIG. 9, the desired functional regions are selected. Such regions include the transcription start site or 5′ cap at step  353 , the 5′ untranslated region at step  354 , the start codon at step  355 , the coding region at step  356 , the stop codon at step  357 , the 3′ untranslated region at step  358 , 5′ splice sites at step  359  or 3′ splice sites at step  360 , specific exons at step  361  or specific introns at step  362 , mRNA stabilization signal at step  363 , mRNA destabilization signal at step  364 , poly-adenylation signal at step  365 , poly-A addition site at step  366 , poly-A tail at step  367 , or the gene sequence 5′ of known pre-mRNA at step  368 . In addition, additional functional sites may be selected, as represented at step  369 .  
     [0111] Many functional regions are important to the proper processing of the gene and are attractive targets for antisense approaches. For example, the AUG start codon is commonly targeted because it is necessary to initiate translation. In addition, splice sites are thought to be attractive targets because these regions are important for processing of the mRNA. Other known sites may be more accessible because of interactions with protein factors or other regulatory molecules.  
     [0112] After the desired functional regions are selected and determined, then a subset of all previously selected oligonucleotides are selected based on hybridization to only those desired functional regions, as represented by step  370 .  
     [0113] 7. Uniform Distribution of Oligonucleotides.  
     [0114] Whether or not targeting functional sites is desired, a large number of oligonucleotide sequences may result from the process thus far. In order to reduce the number of oligonucleotide sequences to a manageable number, a decision is made whether to uniformly distribute selected oligonucleotides along the target, as represented in step  375 . A uniform distribution of oligonucleotide sequences will aim to provide complete coverage throughout the complete target nucleic acid or the selected functional regions. A computer-based program is used to automate the distribution of sequences, as represented in step  380 . Such a program factors in parameters such as length of the target nucleic acid, total number of oligonucleotide sequences desired, oligonucleotide sequences per unit length, number of oligonucleotide sequences per functional region. Manual selection of oligonucleotide sequences is also provided for by step  385 . In some cases, it may be desirable to manually select oligonucleotide sequences. For example, it may be useful to determine the effect of small base shifts on activity. Once the desired number of oligonucleotide sequences is obtained either from step  380  or step  385 , then these oligonucleotide sequences are passed onto step  400  of the process, where oligonucleotide chemistries are assigned.  
     [0115] 8. Assignment of Actual Oligonucleotide Chemistry.  
     [0116] Once a set of select nucleobase sequences has been generated according to the preceding process and decision steps, actual oligonucleotide chemistry is assigned to the sequences. An “actual oligonucleotide chemistry” or simply “chemistry” is a chemical motif that is common to a particular set of robotically synthesized oligonucleotide compounds. Preferred chemistries include, but are not limited to, oligonucleotides in which every linkage is a phosphorothioate linkage, and chimeric oligonucleotides in which a defined number of 5′ and/or 3′ terminal residues have a 2′-methoxyethoxy modification.  
     [0117] Chemistries can be assigned to the nucleobase sequences during general procedure step  400  (FIG. 1). The logical basis for chemistry assignment is illustrated in FIGS. 10 and 11 and an iterative routine for stepping through an oligonucleotide nucleoside by nucleoside is illustrated in FIG. 12. Chemistry assignment can be effected by assignment directly into a word processing program, via an interactive word processing program or via automated programs and devices. In each of these instances, the output file is selected to be in a format that can serve as an input file to automated synthesis devices.  
     [0118] 9. Oligonucleotide Compounds.  
     [0119] In the context of this invention, in reference to oligonucleotides, the term “oligonucleotide” is used to refer to an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or mimetics thereof. Thus this term includes oligonucleotides composed of naturally-occurring nucleobases, sugars and covalent internucleoside (backbone) linkages as well as oligonucleotides having non-naturally-occurring portions which function similarly. Such modified or substituted oligonucleotides are often preferred over native forms, i.e., phosphodiester linked A, C, G, T and U nucleosides, because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target and increased stability in the presence of nucleases.  
     [0120] The oligonucleotide compounds in accordance with this invention can be of various lengths depending on various parameters, including but not limited to those discussed above in reference to the selection criteria of general procedure  300 . For use as antisense oligonucleotides compounds of the invention preferably are from about 8 to about 30 nucleobases in length (i.e. from about 8 to about 30 linked nucleosides). Particularly preferred are antisense oligonucleotides comprising from about 12 to about 25 nucleobases. A discussion of antisense oligonucleotides and some desirable modifications can be found in De Mesmaeker et al.,  Acc. Chem. Res.,  1995, 28, 366. Other lengths of oligonucleotides might be selected for non-antisense targeting strategies, for instance using the oligonucleotides as ribozymes. Such ribozymes normally require oligonucleotides of longer length as is known in the art.  
     [0121] A nucleoside is a base-sugar combination. The base portion of the nucleoside is normally a heterocyclic base. The two most common classes of such heterocyclic bases are the purines and the pyrimidines. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a normal (where normal is defined as being found in RNA and DNA) pentofuranosyl sugar, the phosphate group can be linked to either the 2′, 3′ or 5′ hydroxyl moiety of the sugar. In forming oligonucleotides, the phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound. In turn the respective ends of this linear polymeric structure can be further joined to form a circular structure, however, open linear structures are generally preferred. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide. The normal linkage or backbone of RNA and DNA is a 3′ to 5′ phosphodiester linkage.  
     [0122] Specific examples of preferred oligonucleotides useful in this invention include oligonucleotides containing modified backbones or non-natural internucleoside linkages. As defined in this specification, oligonucleotides having modified backbones include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referenced in the art, modified oligonucleotides that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides.  
     [0123] 10. Selection of Oligonucleotide Chemistries.  
     [0124] In a general logic sheme as illustrated in FIGS. 10 and 11, for each nucleoside position, the user or automated device is interrogated first for a base assignment, followed by a sugar assignment, a linker assignment and finally a conjugate assignment. Thus for each nucleoside, at process step  410  a base is selected. In selecting the base, base chemistry 1 can be selected at process step  412  or one or more alternative bases are selected at process steps  414 ,  416  and  418 . After base selection is effected, the sugar portion of the nucleoside is selected. Thus for each nucleoside, at process step  420  a sugar is selected that together with the select base will complete the nucleoside. In selecting the sugar, sugar chemistry 1 can be selected at process  422  or one or more alternative sugars are selected at process steps  424 ,  426  and  428 . For each two adjacent nucleoside units, at process step  430 , the internucleoside linker is selected. The linker chemistry for the internucleoside linker can be linker chemistry 1 selected at process step  432  or one or more alternative internucleoside linker chemistries are selected at process steps  434 ,  436  and  438 .  
     [0125] In addition to the base, sugar and internucleoside linkage, at each nucleoside position, one or more conjugate groups can be attached to the oligonucleotide via attachment to the nucleoside or attachment to the internucleoside linkage. The addition of a conjugate group is integrated at process step  440  and the assignment of the conjugate group is effected at process step  450 .  
     [0126] For illustrative purposes in FIGS. 10 and 11, for each of the bases, the sugars, the internucleoside linkers, or the conjugates, chemistries 1 though n are illustrated. As described in this specification, it is understood that the number of alternate chemistries between chemistry 1 and alternative chemistry n, for each of the bases, the sugars, the internucleoside linkages and the conjugates, is variable and includes, but is not limited to, each of the specific alternative bases, sugar, internucleoside linkers and conjugates identified in this specification as well as equivalents known in the art.  
     [0127] Utilizing the logic as described in conjunction with FIGS. 10 and 11, chemistry is assigned, as is shown in FIG. 12, to the list of oligonucleotides from general procedure  300 . In assigning chemistries to the oligonucleotides in this list, a pointer can be set at process step  452  to the first oligonucleotide in the list and at step  453  to the first nucleotide of that first oligonucleotide. The base chemistry is selected at step  410 , as described above, the sugar chemistry is selected at step  420 , also as described above, followed by selection of the internucleoside linkage at step  430 , also as described above. At decision  440 , the process branches depending on whether a conjugate will be added at the current nucleotide position. If a conjugate is desired, the conjugate is selected at step  450 , also as described above.  
     [0128] Whether or not a conjugate was added at decision step  440 , an inquiry is made at decision step  454 . This inquiry asks if the pointer resides at the last nucleotide in the current oligonucleotide. If the result at decision step  454  is “No,” the pointer is moved to the next nucleotide in the current oligonucleotide and the loop including steps  410 ,  420 ,  430 ,  440  and  454  is repeated. This loop is reiterated until the result at decision step  454  is “Yes.” 
     [0129] When the result at decision step  454  is “Yes,” a query is made at decision step  460  concerning the location of the pointer in the list of oligonucleotides. If the pointer is not at the last oligonucleotide of the list, the “No” path of the decision step  460  is followed and the pointer is moved to the first nucleotide of the next oligonucleotide in the list at process step  458 . With the pointer set to the next oligonucleotide in the list, the loop that starts at process steps  453  is reiterated. When the result at decision step  460  is “Yes,” chemistry has been assigned to all of the nucleotides in the list of oligonucleotides.  
     [0130] 11. Description of Oligonucleotide Chemistries.  
     [0131] As is illustrated in FIG. 10, for each nucleoside of an oligonucleotide, chemistry selection includes selection of the base forming the nucleoside from a large palette of different base units available. These may be “modified” or “natural” bases (also reference herein as nucleobases) including the natural purine bases adenine (A) and guanine (G), and the natural pyrimidine bases thymine (T), cytosine (C) and uracil (U). They further can include modified nucleobases including other synthetic and natural nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo uracils and cytosines particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine. Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808, those disclosed in the  Concise Encyclopedia Of Polymer Science And Engineering,  pages 858-859, Kroschwitz, J. I., ed. John Wiley &amp; Sons, 1990, those disclosed by Englisch et al.,  Angewandte Chemie, International Edition,  1991, 30, 613, and those disclosed by Sanghvi, Y. S., Chapter 15,  Antisense Research and Applications,  pages 289-302, Crooke, S. T. and Lebleu, B., ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds of the invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., eds.,  Antisense Research and Applications,  CRC Press, Boca Raton, 1993, pp. 276-278) and are presently preferred for selection as the base. These are particularly useful when combined with a 2′-O-methoxyethyl sugar modifications, described below.  
     [0132] Representative United States patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, the above noted U.S. Pat. No. 3,687,808, as well as U.S. Pat. Nos. 4,845,205; 5,130,302; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121, 5,596,091; 5,614,617; and 5,681,941, each of which is incorporated herein by reference in its entirety. Reference is also made to allowed U.S. patent application Ser. No. 08/762,488, filed on Dec. 10, 1996, commonly owned with the present application and which is incorporated herein by reference in its entirety.  
     [0133] In selecting the base for any particular nucleoside of an oligonucleotide, consideration is first given to the need of a base for a particular specificity for hybridization to an opposing strand of a particular target. Thus if an “A” base is required, adenine might be selected however other alternative bases that can effect hybridization in a manner mimicking an “A” base such as 2-aminoadenine might be selected should other consideration, e.g., stronger hybridization (relative to hybridization achieved with adenine), be desired.  
     [0134] As is illustrated in FIG. 10, for each nucleoside of an oligonucleotide, chemistry selection includes selection of the sugar forming the nucleoside from a large palette of different sugar or sugar surrogate units available. These may be modified sugar groups, for instance sugars containing one or more substituent groups. Preferred substituent groups comprise the following at the 2′ position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; or O, S- or N-alkynyl; wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C 1  to C 10  alkyl or C 2  to C 10  alkenyl and alkynyl. Particularly preferred are O[(CH 2 ) n O] m CH 3 , O(CH 2 ) n OCH 3 , O(CH 2 ) n NH 2 , O(CH 2 ) n CH 3 , O(CH 2 ) n ONH 2 , and O(CH 2 ) n ON[(CH 2 ) n CH 3 )] 2 , where n and m are from 1 to about 10. Other preferred substituent groups comprise one of the following at the 2′ position: C 1  to C 10  lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , SO 2 CH 3 , ONO 2 , NO 2 , N 3 , NH 2 , heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties. A preferred modification includes 2′-methoxyethoxy (2′-O—CH 2 CH 2 OCH 3 , also known as 2′-O-(2-methoxyethyl), 2′-O-methoxyethyl, or 2′-MOE) (Martin et al.,  Helv. Chim. Acta,  1995, 78, 486) i.e., an alkoxyalkoxy group. A further preferred modification includes 2′-dimethylamino oxyethoxy, i.e., a O(CH 2 ) 2 ON(CH 3 ) 2  group, also known as 2′-DMAOE, as described in co-owned U.S. patent application Ser. No. 09/016,520, filed on Jan. 30, 1998, which is incorporated herein by reference in its entirety.  
     [0135] Other preferred modifications include 2′-methoxy (2′-O—CH 3 ), 2′-aminopropoxy (2′-OCH 2 CH 2 CH 2 NH 2 ) and 2′-fluoro (2′-F). Similar modifications may also be made at other positions on the sugar group, particularly the 3′ position of the sugar on the 3′ terminal nucleotide or in 2′-5′ linked oligonucleotides and the 5′ position of 5′ terminal nucleotide. The nucleosides of the oligonucleotides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative United States patents that teach the preparation of such modified sugars structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, certain of which are commonly owned with the present application, each of which is incorporated herein by reference in its entirety, together with allowed U.S. patent application Ser. No. 08/468,037, filed on Jun. 5, 1995, which is commonly owned with the present application and which is incorporated herein by reference in its entirety.  
     [0136] As is illustrated in FIG. 10, for each adjacent pair of nucleosides of an oligonucleotide, chemistry selection includes selection of the internucleoside linkage. These internucleoside linkages are also referred to as linkers, backbones or oligonucleotide backbones. For forming these nucleoside linkages, a palette of different internucleoside linkages or backbones is available. These include modified oligonucleotide backbones, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3′-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3′-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalklyphosphotriesters, and boranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′. Various salts, mixed salts and free acid forms are also included.  
     [0137] Representative United States patents that teach the preparation of the above phosphorus containing linkages include, but are not limited to, U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,625,050; and 5,697,248, certain of which are commonly owned with this application, each of which is incorporated herein by reference in its entirety.  
     [0138] Preferred internucleoside linkages for oligonucleotides that do not include a phosphorus atom therein, i.e., for oligonucleosides, have backbones that are formed by short chain alkyl or cycloalkyl intersugar linkages, mixed heteroatom and alkyl or cycloalkyl intersugar linkages, or one or more short chain heteroatomic or heterocyclic intersugar linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH 2  component parts.  
     [0139] Representative United States patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, certain of which are commonly owned with this application, each of which is incorporated herein by reference in its entirety.  
     [0140] In other preferred oligonucleotides, i.e., oligonucleotide mimetics, both the sugar and the intersugar linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar-phosphate backbone of an oligonucleotide is replaced with an amide-containing backbone, in particular an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is incorporated herein by reference in its entirety. Further teaching of PNA compounds can be found in Nielsen et al.,  Science,  1991, 254, 1497.  
     [0141] For the internucleoside linkages, the most preferred embodiments of the invention are oligonucleotides with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular —CH 2 —NH—O—CH 2 —, —CH 2 —N(CH 3 )—O—CH 2 —[known as a methylene (methylimino) or MMI backbone], —CH 2 —O—N(CH 3 )—CH 2 —, —CH 2 —N(CH 3 )—N(CH 3 )—CH 2 — and —O—N(CH 3 )—CH 2 —CH 2 — (wherein the native phosphodiester backbone is represented as —O—P—O—CH 2 —) of the above referenced U.S. Pat. No. 5,489,677, and the amide backbones of the above referenced U.S. Pat. No. 5,602,240. Also preferred are oligonucleotides having morpholino backbone structures of the above-referenced U.S. Pat. No. 5,034,506.  
     [0142] In attaching a conjugate group to one or more nucleosides or internucleoside linkages of an oligonucleotide, various properties of the oligonucleotide are modified. Thus modification of the oligonucleotides of the invention to chemically link one or more moieties or conjugates to the oligonucleotide are intended to enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al.,  Proc. Natl. Acad. Sci. USA,  1989, 86, 6553), cholic acid (Manoharan et al.,  Bioorg. Med. Chem. Let.,  1994, 4, 1053), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al.,  Ann. N.Y Acad. Sci.,  1992, 660,306; Manoharan et al.,  Bioorg. Med. Chem. Let.,  1993, 3, 2765), a thiocholesterol (Oberhauser et al.,  Nucl. Acids Res.,  1992, 20, 533), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al.,  EMBO J.,  1991, 10, 111; Kabanov et al.,  FEBS Lett.,  1990, 259, 327; Svinarchuk et al.,  Biochimie,  1993, 75, 49), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al.,  Tetrahedron Lett.,  1995, 36, 3651; Shea et al.,  Nucl. Acids Res.,  1990, 18, 3777), a polyamine or a polyethylene glycol chain (Manoharan et al.,  Nucleosides  &amp;  Nucleotides,  1995, 14, 969), or adamantane acetic acid (Manoharan et al.,  Tetrahedron Lett.,  1995, 36, 3651), a palmityl moiety (Mishra et al.,  Biochim. Biophys. Acta,  1995, 1264, 229), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al.,  J. Pharmacol. Exp. Ther.,  1996, 277, 923).  
     [0143] Representative United States patents that teach the preparation of such oligonucleotide conjugates include, but are not limited to, U.S. Pat. Nos. 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541,313; 5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,580,731; 5,591,584; 5,109,124; 5,118,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5,245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371,241, 5,391,723; 5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5,565,552; 5,567,810; 5,574,142; 5,585,481; 5,587,371; 5,595,726; 5,597,696; 5,599,923; 5,599,928 and 5,688,941, certain of which are commonly owned with the present application, and each of which is herein incorporated by reference in its entirety.  
     [0144] 12. Chimeric Compounds.  
     [0145] It is not necessary for all positions in a given compound to be uniformly modified. In fact, more than one of the aforementioned modifications may be incorporated in a single compound or even at a single nucleoside within an oligonucleotide. The present invention also includes compounds which are chimeric compounds. “Chimeric” compounds or “chimeras,” in the context of this invention, are compounds, particularly oligonucleotides, which contain two or more chemically distinct regions, each made up of at least one monomer unit, i.e., a nucleotide in the case of an oligonucleotide compound. These oligonucleotides typically contain at least one region wherein the oligonucleotide is modified so as to confer upon the oligonucleotide increased resistance to nuclease degradation, increased cellular uptake, and/or increased binding affinity for the target nucleic acid. An additional region of the oligonucleotide may serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids.  
     [0146] By way of example, RNase H is a cellular endonuclease which cleaves the RNA strand of an RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of oligonucleotide inhibition of gene expression. Consequently, comparable results can often be obtained with shorter oligonucleotides when chimeric oligonucleotides are used, compared to phosphorothioate deoxyoligonucleotides hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art.  
     [0147] Chimeric antisense compounds of the invention may be formed as composite structures representing the union of two or more oligonucleotides, modified oligonucleotides, oligonucleosides and/or oligonucleotide mimetics as described above. Such compounds have also been referred to in the art as “hybrids” or “gapmers”. Representative United States patents that teach the preparation of such hybrid structures include, but are not limited to, U.S. Pat. Nos. 5,013,830; 5,149,797; 5,220,007; 5,256,775; 5,366,878; 5,403,711; 5,491,133; 5,565,350; 5,623,065; 5,652,355; 5,652,356; and 5,700,922, certain of which are commonly owned with the present application and each of which is incorporated herein by reference in its entirety, together with commonly owned and allowed U.S. patent application Ser. No. 08/465,880, filed on Jun. 6, 1995, which is incorporated herein by reference in its entirety.  
     [0148] 13. Description of Automated Oligonucleotide Synthesis.  
     [0149] In the next step of the overall process (illustrated in FIGS. 1 and 2), oligonucleotides are synthesized on an automated synthesizer. Although many devices may be employed, the synthesizer is preferably a variation of the synthesizer described in U.S. Pat. Nos. 5,472,672 and 5,529,756, each of which is incorporated herein by reference in its entirety. The synthesizer described in those patents is modified to include movement in along the Y axis in addition to movement along the X axis. As so modified, a 96-well array of compounds can be synthesized by the synthesizer. The synthesizer further includes temperature control and the ability to maintain an inert atmosphere during all phases of synthesis. The reagent array delivery format employs orthogonal X-axis motion of a matrix of reaction vessels and Y-axis motion of an array of reagents. Each reagent has its own dedicated plumbing system to eliminate the possibility of cross-contamination of reagents and line flushing and/or pipette washing. This in combined with a high delivery speed obtained with a reagent mapping system allows for the extremely rapid delivery of reagents. This further allows long and complex reaction sequences to be performed in an efficient and facile manner.  
     [0150] The software that operates the synthesizer allows the straightforward programming of the parallel synthesis of a large number of compounds. The software utilizes a general synthetic procedure in the form of a command (.cmd) file, which calls upon certain reagents to be added to certain wells via lookup in a sequence (.seq) file. The bottle position, flow rate, and concentration of each reagent is stored in a lookup table (.tab) file. Thus, once any synthetic method has been outlined, a plate of compounds is made by permutating a set of reagents, and writing the resulting output to a text file. The text file is input directly into the synthesizer and used for the synthesis of the plate of compounds. The synthesizer is interfaced with a relational database allowing data output related to the synthesized compounds to be registered in a highly efficient manner.  
     [0151] Building of the .seq, .cmd and .tab files is illustrated in FIG. 13. Thus as a part of the general oligonucleotide synthesis procedure  500 , for each linker chemistry at process step  502 , a synthesis file, i.e., a .cmd file, is built at process step  504 . This file can be built fresh to reflect a completely new set of machine commands reflecting a set of chemical synthesis steps or it can modify an existing file stored at process step  504  by editing that stored file in process step  508 . The .cmd files are built using a word processor and a command set of instructions as outlined below.  
     [0152] It will be appreciated that the preparation of control software and data files is within the routine skill of persons skilled in annotated nucleotide synthesis. The same will depend upon the hardware employed, the chemistries adopted and the design paradigm selected by the operator.  
     [0153] In a like manner to the building the .cmd files, .tab files are built to reflect the necessary reagents used in the automatic synthesizer for the particular chemistries that have been selected for the linkages, bases, sugars and conjugate chemistries. Thus for each of a set of these chemistries at process step  510 , a .tab file is built at process step  512  and stored at process step  514 . As with the .cmd files, an existing .tab file can be edited at process step  516 .  
     [0154] Both the .cmd files and the .tab files are linked together at process step  518  and stored for later retrieval in an appropriate sample database  520 . Linking can be as simple as using like file names to associate a .cmd file to its appropriate .tab file, e.g., synthesis — 1.cmd is linked to synthesis — 1.tab by use of the same preamble in their names.  
     [0155] The automated, multi-well parallel array synthesizer employs a reagent array delivery format, in which each reagent utilized has a dedicated plumbing system. As seen in FIGS. 23 and 24, an inert atmosphere  522  is maintained during all phases of a synthesis. Temperature is controlled via a thermal transfer plate  524 , which holds an injection molded reaction block  526 . The reaction plate assembly slides in the X-axis direction, while for example eight nozzle blocks ( 528 ,  530 ,  532 ,  534 ,  536 ,  538 ,  540  and  542 ) holding the reagent lines slide in the Y-axis direction, allowing for the extremely rapid delivery of any of 64 reagents to 96 wells. In addition, there are for example, six banks of fixed nozzle blocks ( 544 ,  546 ,  548 ,  550 ,  552  and  554 ) which deliver the same reagent or solvent to eight wells at once, for a total of 72 possible reagents.  
     [0156] In synthesizing oligonucleotides for screening, the target reaction vessels, a 96 well plate  556  (a 2-dimensional array), moves in one direction along the X axis, while the series of independently controlled reagent delivery nozzles ( 528 ,  530 ,  532 ,  534 ,  536 ,  538 ,  540  and  542 ) move along the Y-axis relative to the reaction vessel  558 . As the reaction plate  556  and reagent nozzles ( 528 ,  530 ,  532 ,  534 ,  536 ,  538 ,  540  and  542 ) can be moved independently at the same time, this arrangement facilitates the extremely rapid delivery of up to 72 reagents independently to each of the 96 reaction vessel wells.  
     [0157] The system software allows the straightforward programming of the synthesis of a large number of compounds by supplying the general synthetic procedure in the form of the command file to call upon certain reagents to be added to specific wells via lookup in the sequence file with the bottle position, flow rate, and concentration of each reagent being stored in the separate reagent table file. Compounds can be synthesized on various scales. For oligonucleotides, a 200 nmole scale is typically selected while for other compounds larger scales, as for example a 10 μmole scale (3-5 mg), might be utilized. The resulting crude compounds are generally &gt;80% pure, and are utilized directly for high throughput screening assays. Alternatively, prior to use the plates can be subjected to quality control (see general procedure 600 and Example 8) to ascertain their exact purity. Use of the synthesizer results in a very efficient means for the parallel synthesis of compounds for screening.  
     [0158] The software inputs accept tab delimited text files (as discussed above for file  504  and  512 ) from any text editor. A typical command file, a .cmd file, is shown in Example 3 at Table 2. Typical sequence files, .seq files, are shown in Example 3 at Tables 3 and 4 (.SEQ file), and a typical reagent file, a .tab file, is shown in Example 3 at Table 5. Table 3 illustrates the sequence file for an oligonucleotide having 2′-deoxy nucleotides at each position with a phosphorothioate backbone throughout. Table 4 illustrates the sequence file for an oligonucleotide, again having a phosphorothioate backbone throughout, however, certain modified nucleoside are utilized in portions of the oligonucleotide. As shown in this table, 2′-O-(2-methoxyethyl) modified nucleosides are utilized in a first region (a wing) of the oligonucleotide, followed by a second region (a gap) of 2′-deoxy nucleotides and finally a third region (a further wing) that has the same chemistry as the first region. Typically some of the wells of the 96 well plate  556  may be left empty (depending on the number of oligonucleotides to be made during an individual synthesis) or some of the wells may have oligonucleotides that will serve as standards for comparison or analytical purposes.  
     [0159] Prior to loading reagents, moisture sensitive reagent lines are purged with argon at  522  for 20 minutes. Reagents are dissolved to appropriate concentrations and installed on the synthesizer. Large bottles, collectively identified as  558  in FIG. 23 (containing 8 delivery lines) are used for wash solvents and the delivery of general activators, trityl group cleaving reagents and other reagents that may be used in multiple wells during any particular synthesis. Small septa bottles, collectively identified as  560  in FIG. 23, are utilized to contain individual nucleotide amidite precursor compounds. This allows for anhydrous preparation and efficient installation of multiple reagents by using needles to pressurize the bottle, and as a delivery path. After all reagents are installed, the lines are primed with reagent, flow rates measured, then entered into the reagent table (.tab file). A dry resin loaded plate is removed from vacuum and installed in the machine for the synthesis.  
     [0160] The modified 96 well polypropylene plate  556  is utilized as the reaction vessel. The working volume in each well is approximately 700 μl. The bottom of each well is provided with a pressed-fit 20 μm polypropylene frit and a long capillary exit into a lower collection chamber as is illustrated in FIG. 5 of the above referenced U.S. Pat. No. 5,372,672. The solid support for use in holding the growing oligonucleotide during synthesis is loaded into the wells of the synthesis plate  556  by pipetting the desired volume of a balanced density slurry of the support suspended in an appropriate solvent, typically an acetonitrile-methylene chloride mixture. Reactions can be run on various scales as for instance the above noted 200 nmole and 10 μmol scales. For oligonucleotide synthesis a CPG support is preferred, however other medium loading polystyrene-PEG supports such as TENTAGEL™ or ARGOGEL™ can also be used.  
     [0161] As seen in FIG. 24, the synthesis plate is transported back and forth in the X-direction under an array of 8 moveable banks ( 530 ,  532 ,  534 ,  536 ,  538 ,  540 ,  542  and  544 ) of 8 nozzles (64 total) in the Y-direction, and 6 banks ( 544 ,  546 ,  548 ,  550 ,  552  and  554 ) of 48 fixed nozzles, so that each well can receive the appropriate amounts of reagents and/or solvents from any reservoir (large bottle or smaller septa bottle). A sliding balloon-type seal  562  surrounds this nozzle array and joins it to the reaction plate headspace  564 . A slow sweep of nitrogen or argon  522  at ambient pressure across the plate headspace is used to preserve an anhydrous environment.  
     [0162] The liquid contents in each well do not drip out until the headspace pressure exceeds the capillary forces on the liquid in the exit nozzle. A slight positive pressure in the lower collection chamber can be added to eliminate residual slow leakage from filled wells, or to effect agitation by bubbling inert gas through the suspension. In order to empty the wells, the headspace gas outlet valve is closed and the internal pressure raised to about 2 psi. Normally, liquid contents are blown directly to waste  566 . However, a 96 well microtiter plate can be inserted into the lower chamber beneath the synthesis plate in order to collect the individual well eluents for spectrophotometric monitoring (trityl, etc.) of reaction progress and yield.  
     [0163] The basic plumbing scheme for the machine is the gas-pressurized delivery of reagents. Each reagent is delivered to the synthesis plate through a dedicated supply line, collectively identified at  568 , solenoid valve collectively identified at  570  and nozzle, collectively identified at  572 . Reagents never cross paths until they reach the reaction well. Thus, no line needs to be washed or flushed prior to its next use and there is no possibility of cross-contamination of reagents. The liquid delivery velocity is sufficiently energetic to thoroughly mix the contents within a well to form a homogeneous solution, even when employing solutions having drastically different densities. With this mixing, once reactants are in homogeneous solution, diffusion carries the individual components into and out of the solid support matrix where the desired reaction takes place. Each reagent reservoir can be plumbed to either a single nozzle or any combination of up to 8 nozzles. Each nozzle is also provided with a concentric nozzle washer to wash the outside of the delivery nozzles in order to eliminate problems of crystallized reactant buildup due to slow evaporation of solvent at the tips of the nozzles. The nozzles and supply lines can be primed into a set of dummy wells directly to waste at any time.  
     [0164] The entire plumbing system is fabricated with teflon tubing, and reagent reservoirs are accessed via syringe needle/septa or direct connection into the higher capacity bottles. The septum vials  560  are held in removable 8-bottle racks to facilitate easy setup and cleaning. The priming volume for each line is about 350 μl. The minimum delivery volume is about 2 μl, and flow rate accuracy is ±5%. The actual amount of material delivered depends on a timed flow of liquid. The flow rate for a particular solvent will depend on its viscosity and wetting characteristics of the teflon tubing. The flow rate (typically 200-350 μl per sec) is experimentally determined, and this information is contained in the reagent table setup file.  
     [0165] Heating and cooling of the reaction block  526  is effected utilizing a recirculating heat exchanger plate  524 , similar to that found in PCR thermocyclers, that nests with the polypropylene synthesis plate  556  to provide good thermal contact. The liquid contents in a well can be heated or cooled at about 10° C. per minute over a range of +5 to +80° C., as polypropylene begins to soften and deform at about 80° C. For temperatures greater than this, a non-disposable synthesis plate machined from stainless steel or monel with replaceable frits can be utilized.  
     [0166] The hardware controller can be any of a wide variety, but conveniently can be designed around a set of three 1 MHz 86332 chips. This controller is used to drive the single X-axis and 8 Y-axis stepper motors as well as provide the timing functions for a total of 154 solenoid valves. Each chip has 16 bidirectional timer I/O and 8 interrupt channels in its timer processing unit (TPU). These are used to provide the step and direction signals, and to read 3 encoder inputs and 2 limit switches for controlling up to three motors per chip. Each 86332 chip also drives a serial chain of 8 UNC5891A darlington array chips to provide power to 64 valves with msec resolution. The controller communicates with the Windows software interface program running on a PC via a 19200 Hz serial channel, and uses an elementary instruction set to communicate valve_number, time_open, motor_number and position_data.  
     [0167] The three components of the software program that run the array synthesizer are the generalized procedure or command (.cmd) file which specifies the synthesis instructions to be performed, the sequence (.seq) file which specifies the scale of the reaction and the order in which variable groups will be added to the core synthon, and the reagent table (.tab) file which specifies the name of a chemical, its location (bottle number), flow rate, and concentration are utilized in conjunction with a basic set of command instructions.  
     [0168] One basic set of command instructions can be:  
                                                  ADD                                 IF   {block of instructions}   END_IF           REPEAT   {block of instructions}   END_REPEAT                         PRIME, NOZZLE_WASH           WAIT, DRAIN           LOAD, REMOVE           NEXT_SEQUENCE           LOOP_BEGIN, LOOP_END                      
 
     [0169] The ADD instruction has two forms, and is intended to have the look and feel of a standard chemical equation. Reagents are specified to be added by a molar amount if the number proceeds the name identifier, or by an absolute volume in microliters if the number follows the identifier. The number of reagents to be added is a parsed list, separated by the “+” sign. For variable reagent identifiers, the key word, &lt;seq&gt;, means look in the sequence table for the identity of the reagent to be added, while the key word, &lt;act&gt;, means add the reagent which is associated with that particular &lt;seq&gt;. Reagents are delivered in the order specified in the list.  
     [0170] Thus:  
     [0171] ADD ACN 300  
     [0172] means: Add 300 μl of the named reagent acetonitrile; ACN to each well of active synthesis  
     [0173] ADD &lt;seq&gt;300  
     [0174] means: If the sequence pointer in the .seq file is to a reagent in the list of reagents, independent of scale, add 300 μl of that particular reagent specified for that well.  
     [0175] ADD 1.1 PYR+1.0&lt;seq&gt;+1.1&lt;act1&gt; 
     [0176] means: If the sequence pointer in the .seq file is to a reagent in the list of acids in the Class ACIDS — 1, and PYR is the name of pyridine, and ethyl chloroformate is defined in the .tab file to activate the class, ACIDS — 1, then this instruction means:  
     [0177] ADD 1.1 equiv. pyridine  
     [0178] 1.0 equiv. of the acid specified for that well and  
     [0179] 1.1 equiv. of the activator, ethyl chloroformate  
     [0180] The IF command allows one to test what type of reagent is specified in the &lt;seq&gt;variable and process the succeeding block of commands accordingly.  
                                                  Thus:                             ACYLATION   {the procedure name}                         BEGIN                         IF CLASS = ACIDS_1                         ADD 1.0 &lt;seq&gt; + 1.1 &lt;act1&gt; + 1.1 PYR           WAIT 60                         ENDIF           IF CLASS = ACIDS_2                         ADD 1.0 &lt;seq&gt; + 1.2 &lt;act1&gt; + 1.2 TEA                         ENDIF           WAIT 60           DRAIN 10                         END                      
 
     [0181] means: Operate on those wells for which reagents contained in the Acid — 1 class are specified, WAIT 60 sec, then operate on those wells for which reagents contained in the Acid — 2 class are specified, then WAIT 60 sec longer, then DRAIN the whole plate. Note that the Acid — 1 group has reacted for a total of 120 sec, while the Acid — 2 group has reacted for only 60 sec.  
     [0182] The REPEAT command is a simple way to execute the same block of commands multiple times.  
                                                  Thus:                             WASH_1   {the procedure name}                         BEGIN                         REPEAT 3                         ADD ACN 300           DRAIN 15                         END_REPEAT                         END                      
 
     [0183] means: repeats the add acetonitrile and drain sequence for each well three times.  
     [0184] The PRIME command will operate either on specific named reagents or on nozzles which will be used in the next associated &lt;seq&gt;operation. The μl amount dispensed into a prime port is a constant that can be specified in a config.dat file.  
     [0185] The NOZZLE_WASH command for washing the outside of reaction nozzles free from residue due to evaporation of reagent solvent will operate either on specific named reagents or on nozzles which have been used in the preceding associated &lt;seq&gt;operation. The machine is plumbed such that if any nozzle in a block has been used, all the nozzles in that block will be washed into the prime port.  
     [0186] The WAIT and DRAIN commands are by seconds, with the drain command applying a gas pressure over the top surface of the plate in order to drain the wells.  
     [0187] The LOAD and REMOVE commands are instructions for the machine to pause for operator action.  
     [0188] The NEXT_SEQUENCE command increments the sequence pointer to the next group of substituents to be added in the sequence file. The general form of a .seq file entry is the definition:  
     Well_No Well_ID Scale Sequence  
     [0189] The sequence information is conveyed by a series of columns, each of which represents a variable reagent to be added at a particular position. The scale (μmole) variable is included so that reactions of different scale can be run at the same time if desired. The reagents are defined in a lookup table (the .tab file), which specifies the name of the reagent as referred to in the sequence and command files, its location (bottle number), flow rate, and concentration. This information is then used by the controller software and hardware to determine both the appropriate slider motion to position the plate and slider arms for delivery of a specific reagent, as well as the specific valve and time required to deliver the appropriate reagents. The adept classification of reagents allows the use of conditional IF loops from within a command file to perform addition of different reagents differently during a “single step” performed across 96 wells simultaneously. The special class ACTIVATORS defines certain reagents that always get added with a particular class of reagents (for example tetrazole during a phosphitylation reaction in adding the next nucleotide to a growing oligonucleotide).  
     [0190] The general form of the .tab file is the definition:  
     Class Bottle Reagent Name Flow_rate Conc.  
     [0191] The LOOP_BEGIN and LOOP_END commands define the block of commands which will continue to operate until a NEXT_SEQUENCE command points past the end of the longest list of reactants in any well.  
     [0192] Not included in the command set is a MOVE command. For all of the above commands, if any plate or nozzle movement is required, this is automatically executed in order to perform the desired solvent or reagent delivery operation. This is accomplished by the controller software and hardware, which determines the correct nozzle(s) and well(s) required for a particular reagent addition, then synchronizes the position of the requisite nozzle and well prior to adding the reagent.  
     [0193] A MANUAL mode can also be utilized in which the synthesis plate and nozzle blocks can be “homed” or moved to any position by the operator, the nozzles primed or washed, the various reagent bottles depressurized or washed with solvent, the chamber pressurized, etc. The automatic COMMAND mode can be interrupted at any point, MANUAL commands executed, and then operation resumed at the appropriate location. The sequence pointer can be incremented to restart a synthesis anywhere within a command file.  
     [0194] In reference to FIG. 14, the list of oligonucleotides for synthesis can be rearranged or grouped for optimization of synthesis. Thus at process step  574 , the oligonucleotides are grouped according to a factor on which to base the optimization of synthesis. As illustrated in the Examples below, one such factor is the 3′ most nucleoside of the oligonucleotide. Using the amidite approach for oligonucleotide synthesis, a nucleotide bearing a 3′ phosphoramite is added to the 5′ hydroxyl group of a growing nucleotide chain. The first nucleotide (at the 3′ terminus of the oligonucleotide—the 3′ most nucleoside) is first connected to a solid support. This is normally done batchwise on a large scale as is standard practice during oligonucleotide synthesis.  
     [0195] Such solid supports pre-loaded with a nucleoside are commercially available. In utilizing the multi well format for oligonucleotide synthesis, for each oligonucleotide to be synthesized, an aliquot of a solid support bearing the proper nucleoside thereon is added to the well for synthesis. Prior to loading the sequence of oligonucleotides to be synthesized in the .seq file, they are sorted by the 3′ terminal nucleotide. Based on that sorting, all of the oligonucleotide sequences having an “A” nucleoside at their 3′ end are grouped together, those with a “C” nucleoside are grouped together as are those with “G” or “T” nucleosides. Thus in loading the nucleoside-bearing solid support into the synthesis wells, machine movements are conserved.  
     [0196] The oligonucleotides can be grouped by the above described parameter or other parameters that facilitate the synthesis of the oligonucleotides. Thus in FIG. 14, sorting is noted as being effected by some parameter of type 1, as for instance the above described 3′ most nucleoside, or other types of parameters from type 2 to type n at process steps  576 ,  578  and  580 . Since synthesis will be from the 3′ end of the oligonucleotides to the 5′ end, the oligonucleotide sequences are reverse sorted to read 3′ to 5′. The oligonucleotides are entered in the .seq file in this form, i.e., reading 3′ to 5′.  
     [0197] Once sorted into types, the position of the oligonucleotides on the synthesis plates is specified at process step  582  by the creation of a .seq file as described above. The .seq file is associated with the respective .cmd and .tab files needed for synthesis of the particular chemistries specified for the oligonucleotides at process step  584  by retrieval of the .cmd and .tab files at process step  586  from the sample database  520 . These files are then input into the multi well synthesizer at process step  588  for oligonucleotide synthesis. Once physically synthesized, the list of oligonucleotides again enters the general procedure flow as indicated in FIG. 1. For shipping, storage or other handling purposes, the plates can be lyophilized at this point if desired. Upon lyophilization, each well contains the oligonucleotides located therein as a dry compound.  
     [0198] 14. Quality Control.  
     [0199] In an optional step, quality control is performed on the oligonucleotides at process step  600  after a decision is made (decision step  550 ) to perform quality control. Although optional, quality control may be desired when there is some reason to think that some aspect of the synthetic process step  500  has been compromised. Alternatively, samples of the oligonucleotides may be taken and stored in the event that the results of assays conducted using the oligonucleotides (process step  700 ) yield confusing results or suboptimal data. In the latter event, for example, quality control might be performed after decision step  800  if no oligonucleotides with sufficient activity are identified. In either event, decision step  650  follows quality control step process  600 . If one or more of the oligonucleotides do not pass quality control, process step  500  can be repeated, i.e., the oligonucleotides are synthesized for a second time.  
     [0200] The operation of the quality control system general procedure  600  is detailed in steps  610 - 660  of FIG. 15. Also referenced in the following discussion are the robotics and associated analytical instrumentation as shown in FIG. 18.  
     [0201] During step  610  (FIG. 15), sterile, double-distilled water is transferred by an automated liquid handler ( 2040  of FIG. 18) to each well of a multi-well plate containing a set of lyophilized antisense oligonucleotides. The automated liquid handler ( 2040  of FIG. 18) reads the barcode sticker on the multi-well plate to obtain the plate&#39;s identification number. Automated liquid handler  2040  then queries Sample Database  520  (which resides in Database Server  2002  of FIG. 18) for the quality control assay instruction set for that plate and executes the appropriate steps. Three quality control processes are illustrated, however, it is understood that other quality control processes or steps maybe practiced in addition to or in place of the processes illustrated.  
     [0202] The first illustrative quality control process (steps  622  to  626 ) quantitates the concentration of oligonucleotide in each well. If this quality control step is performed, an automated liquid handler ( 2040  of FIG. 18) is instructed to remove an aliquot from each well of the master plate and generate a replicate daughter plate for transfer to the UV spectrophotometer ( 2016  of FIG. 18). The UV spectrophotometer ( 2016  of FIG. 18) then measures the optical density of each well at a wavelength of 260 nanometers. Using standardized conversion factors, a microprocessor within UV spectrophotometer ( 2016  of FIG. 18) then calculates a concentration value from the measured absorbance value for each well and output the results to Sample Database  520 .  
     [0203] The second illustrative quality control process steps  632  to  636 ) quantitates the percent of total oligonucleotide in each well that is full length. If this quality control step is performed, an automated liquid handler ( 2040  of FIG. 18) is instructed to remove an aliquot from each well of the master plate and generate a replicate daughter plate for transfer to the multichannel capillary gel electrophoresis apparatus ( 2022  of FIG. 18). The apparatus electrophoretically resolves in capillary tube gels the oligonucleotide product in each well. As the product reaches the distal end of the tube gel during electrophoresis, a detection window dynamically measures the optical density of the product that passes by it. Following electrophoresis, the value of percent product that passed by the detection window with respect to time is utilized by a built in microprocessor to calculate the relative size distribution of oligonucleotide product in each well. These results are then output to the Sample Database ( 520 .  
     [0204] The third illustrative quality control process steps  632  to  636 ) quantitates the mass of the oligonucleotide in each well that is full length. If this quality control step is performed, an automated liquid handler ( 2040  of FIG. 18) is instructed to remove an aliquot from each well of the master plate and generate a replicate daughter plate for transfer to the multichannel liquid electrospray mass spectrometer ( 2018  of FIG. 18). The apparatus then uses electrospray technology to inject the oligonucleotide product into the mass spectrometer. A built in microprocessor calculates the mass-to-charge ratio to arrive at the mass of oligonucleotide product in each well. The results are then output to Sample Database  520 .  
     [0205] Following completion of the selected quality control processes, the output data is manually examined or is examined using an appropriate algorithm and a decision is made as to whether or not the plate receives “Pass” or “Fail” status. The current criteria for acceptance, for 18 mer oligonucleotides, is that at least 85% of the oligonucleotides in a multi-well plate must be 85% or greater full length product as measured by both capillary gel electrophoresis and mass spectrometry. An input (manual or automated) is then made into Sample Database  520  as to the pass/fail status of the plate. If a plate fails, the process cycles back to step  500 , and a new plate of the same oligonucleotides is automatically placed in the plate synthesis request queue (process  554  of FIG. 15). If a plate receives “Pass” status, an automated liquid handler ( 2040  of FIG. 18) is instructed to remove appropriate aliquots from each well of the master plate and generate two replicate daughter plates in which the oligonucleotide in each well is at a concentration of 30 micromolar. The plate then moves on to process  700  for oligonucleotide activity evaluation.  
     [0206] 15. Cell Lines for Assaying Oligonucleotide Activity.  
     [0207] The effect of antisense compounds on target nucleic acid expression can be tested in any of a variety of cell types provided that the target nucleic acid, or its gene product, is present at measurable levels. This can be routinely determined using, for example, PCR or Northern blot analysis. The following four cell types are provided for illustrative purposes, but other cell types can be routinely used.  
     [0208] T-24 cells: The transitional cell bladder carcinoma cell line T-24 is obtained from the American Type Culture Collection (ATCC) (Manassas, Va.). T-24 cells were routinely cultured in complete McCoy&#39;s 5A basal media (Life Technologies, Gaithersburg, Md.) supplemented with 10% fetal calf serum, penicillin 100 units per milliliter, and streptomycin 100 micrograms per milliliter (all from Life Technologies). Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence. Cells are routinely seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis. For Northern blotting or other analysis, cells are seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.  
     [0209] A549 cells: The human lung carcinoma cell line A549 is obtained from the ATCC (Manassas, Va.). A549 cells were routinely cultured in DMEM basal media (Life Technologies) supplemented with 10% fetal calf serum, penicillin 100 units per milliliter, and streptomycin 100 micrograms per milliliter (all from Life Technologies). Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence.  
     [0210] NHDF cells: Human neonatal dermal fibroblast (NHDF) were obtained from the Clonetics Corporation (Walkersville, Md.). NHDFs were routinely maintained in Fibroblast Growth Medium (Clonetics Corp.) as provided by the supplier. Cells are maintained for up to 10 passages as recommended by the supplier.  
     [0211] HEK cells: Human embryonic keratinocytes (HEK) were obtained from the Clonetics Corp. HEKs were routinely maintained in Keratinocyte Growth Medium (Clonetics Corp.) as provided by the supplier. Cell are routinely maintained for up to 10 passages as recommended by the supplier.  
     [0212] 16. Treatment of Cells with Candidate Compounds:  
     [0213] When cells reach about 80% confluency, they are treated with oligonucleotide. For cells grown in 96-well plates, wells are washed once with 200 μl OPTI-MEM-1™ reduced-serum medium (Life Technologies) and then treated with 130 μl of OPTI-MEM-1™ containing 3.75 μg/ml LIPOFECTIN™ (Life Technologies) and the desired oligonucleotide at a final concentration of 150 nM. After 4 hours of treatment, the medium was replaced with fresh medium. Cells were harvested 16 hours after oligonucleotide treatment.  
     [0214] Alternatively, for cells resistant to cationic mediated transfection, oligonucleotides can be introduced by electroporation. Electroporation conditions must be optimized for every cell type. In general, oligonucleotide is added directly to complete growth media to a final concentration between 1 and 20 micromolar. An electronic pulse is delivered to the cells using a BTX T820 ELECTRO SQUARE PORATOR™ using a Multi-coaxial 96-well electrode (BT840) (BTX Corporation, San Diego, Calif.). Following electroporation, the cells are returned to the incubator for 16 hours.  
     [0215] 17. Assaying Oligonucleotide Activity:  
     [0216] Oligonucleotide-mediated modulation of expression of a target nucleic acid can be assayed in a variety of ways known in the art. For example, target RNA levels can be quantitated by, e.g., Northern blot analysis, competitive PCR, or reverse transcriptase polymerase chain reaction (RT-PCR). RNA analysis can be performed on total cellular RNA or, preferably in the case of polypeptide-encoding nucleic acids, poly(A)+ mRNA. For RT-PCR, poly(A)+ mRNA is preferred. Methods of RNA isolation are taught in, for example, Ausubel et al. ( Short Protocols in Molecular Biology,  2nd Ed., pp. 4-1 to 4-13, Greene Publishing Associates and John Wiley &amp; Sons, New York, 1992). Northern blot analysis is routine in the art (Id., pp. 4-14 to 4-29).  
     [0217] Alternatively, total RNA can be prepared from cultured cells or tissue using the QIAGEN RNeasy®-96 kit for the high throughput preparation of RNA (QIAGEN, Inc., Valencia, Calif.). Essentially, protocols are carried out according to the manufacturer&#39;s directions. Optionally, a DNase step is included to remove residual DNA prior to RT-PCR.  
     [0218] To improve efficiency and accuracy the repetitive pipeting steps and elution step have been automated using a QIAGEN Bio-Robot 9604. Essentially after lysing of the oligonucleotide treated cell cultures in situ, the plate is transferred to the robot deck where the pipeting, DNase treatment, and elution steps are carried out.  
     [0219] Reverse transcriptase polymerase chain reaction (RT-PCR) can be conveniently accomplished using the commercially available ABI PRISM® 7700 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to the manufacturer&#39;s instructions. Other methods of PCR are also known in the art.  
     [0220] Target protein levels can be quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), Enzyme-linked immunosorbent assay (ELISA) or fluorescence-activated cell sorting (FACS). Antibodies directed to a protein encoded by a target nucleic acid can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies, (Aerie Corporation, Birmingham, Mich. or via the world wide web of the interact at ANTIBODIES-PROBES.com/), or can be prepared via conventional antibody generation methods. Methods for preparation of polyclonal, monospecific (“antipeptide”) and monoclonal antisera are taught by, for example, Ausubel et al. ( Short Protocols in Molecular Biology,  2nd Ed., pp. 11-3 to 11-54, Greene Publishing Associates and John Wiley &amp; Sons, New York, 1992).  
     [0221] Immunoprecipitation methods are standard in the art and are described by, for example, Ausubel et al. (Id., pp. 10-57 to 10-63). Western blot (immunoblot) analysis is standard in the art (Id., pp. 10-32 to 10-10-35). Enzyme-linked immunosorbent assays (ELISA) are standard in the art (Id., pp. 11-5 to 11-17).  
     [0222] Because it is preferred to assay the compounds of the invention in a batchwise fashion, i.e., in parallel to the automated synthesis process described above, preferred means of assaying are suitable for use in 96-well plates and with robotic means. Accordingly, automated RT-PCR is preferred for assaying target nucleic acid levels, and automated ELISA is preferred for assaying target protein levels.  
     [0223] The assaying step, general procedure step  700 , is described in detail in FIG. 16. After an appropriate cell line is selected at process step  710 , a decision is made at decision step  714  as to whether RT-PCR will be the only method by which the activity of the compounds is evaluated. In some instances, it is desirable to run alternative assay methods at process step  718 ; for example, when it is desired to assess target polypeptide levels as well as target RNA levels, an immunoassay such as an ELISA is run in parallel with the RT-PCR assays. Preferably, such assays are tractable to semi-automated or robotic means.  
     [0224] When RT-PCR is used to evaluate the activities of the compounds, cells are plated into multi-well plates (typically, 96-well plates) in process step  720  and treated with test or control oligonucleotides in process step  730 . Then, the cells are harvested and lysed in process step  740  and the lysates are introduced into an apparatus where RT-PCR is carried out in process step  750 . A raw data file is generated, and the data is downloaded and compiled at step  760 . Spreadsheet files with data charts are generated at process step  770 , and the experimental data is analyzed at process step  780 . Based on the results, a decision is made at process step  785  as to whether it is necessary to repeat the assays and, if so, the process begins again with step  720 . In any event, data from all the assays on each oligonucleotide are compiled and statistical parameters are automatically determined at process step  790 .  
     [0225] 18. Classification of Compounds Based on their Activity:  
     [0226] Following assaying, general procedure step  700 , oligonucleotide compounds are classified according to one or more desired properties. Typically, three classes of compounds are used: active compounds, marginally active (or “marginal”) compounds and inactive compounds. To some degree, the selection criteria for these classes vary from target to target, and members of one or more classes may not be present for a given set of oligonucleotides.  
     [0227] However, some criteria are constant. For example, inactive compounds will typically comprise those compounds having 5% or less inhibition of target expression (relative to basal levels). Active compounds will typically cause at least 30% inhibition of target expression, although lower levels of inhibition are acceptable in some instances. Marginal compounds will have activities intermediate between active and inactive compounds, with preferred marginal compounds having activities more like those of active compounds.  
     [0228] 19. Optimization of Lead Compounds by Sequence.  
     [0229] One means by which oligonucleotide compounds are optimized for activity is by varying their nucleobase sequences so that different regions of the target nucleic acid are targeted. Some such regions will be more accessible to oligonucleotide compounds than others, and “sliding” a nucleobase sequence along a target nucleic acid only a few bases can have significant effects on activity. Accordingly, varying or adjusting the nucleobase sequences of the compounds of the invention is one means by which suboptimal compounds can be made optimal, or by which new active compounds can be generated.  
     [0230] The operation of the gene walk process  1100  detailed in steps  1104 - 1112  of FIG. 17 is detailed as follows. As used herein, the term “gene walk” is defined as the process by which a specified oligonucleotide sequence x that binds to a specified nucleic acid target y is used as a frame of reference around which a series of new oligonucleotides sequences capable of hybridizing to nucleic acid target y are generated that are sequence shifted increments of oligonucleotide sequence x. Gene walking can be done “downstream”, “upstream” or in both directions from a specified oligonucleotide.  
     [0231] During step  1104  the user manually enters the identification number of the oligonucleotide sequence around which it is desired to execute gene walk process  1100  and the name of the corresponding target nucleic acid. The user then enters the scope of the gene walk at step  1104 , by which is meant the number of oligonucleotide sequences that it is desired to generate. The user then enters in step  1108  a positive integer value for the sequence shift increment. Once this data is generated, the gene walk is effected. This causes a subroutine to be executed that automatically generates the desired list of sequences by walking along the target sequence. At that point, the user proceeds to process  400  to assign chemistries to the selected oligonucleotides.  
     [0232] Example 18 below, details a gene walk. In subsequent steps, this new set of nucleobase sequences generated by the gene walk is used to direct the automated synthesis at general procedure step  500  of a second set of candidate oligonucleotides. These compounds are then taken through subsequent process steps to yield active compounds or reiterated as necessary to optimize activity of the compounds.  
     [0233] 20. Optimization of Lead Compounds by Chemistry.  
     [0234] Another means by which oligonucleotide compounds of the invention are optimized is by reiterating portions of the process of the invention using marginal or active compounds from the first iteration and selecting additional chemistries to the nucleobase sequences thereof.  
     [0235] Thus, for example, an oligonucleotide chemistry different from that of the first set of oligonucleotides is assigned at general procedure step  400 . The nucleobase sequences of marginal compounds are used to direct the synthesis at general procedure step  500  of a second set of oligonucleotides having the second assigned chemistry. The resulting second set of oligonucleotide compounds is assayed in the same manner as the first set at procedure process step  700  and the results are examined to determine if compounds having sufficient activity have been generated at decision step  800 .  
     [0236] 21. Identification of Sites Amenable to Antisense Technologies.  
     [0237] In a related process, a second oligonucleotide chemistry is assigned at procedure step  400  to the nucleobase sequences of all of the oligonucleotides (or, at least, all of the active and marginal compounds) and a second set of oligonucleotides is synthesized at procedure step  500  having the same nucleobase sequences as the first set of compounds. The resulting second set of oligonucleotide compounds is assayed in the same manner as the first set at procedure step  700  and active and marginal compounds are identified at procedure steps  800  and  1000 .  
     [0238] In order to identify sites on the target nucleic acid that are amenable to a variety of antisense technologies, the following mathematically simple steps are taken. The sequences of active and marginal compounds from two or more such automated syntheses/assays are compared and a set of nucleobase sequences that are active, or marginally so, in both sets of compounds is identified. The reverse complements of these nucleobase sequences corresponds to sequences of the target nucleic acid that are tractable to a variety of antisense and other sequence-based technologies. These antisense-sensitive sites are assembled into contiguous sequences (contigs) using the procedures described for assembling target nucleotide sequences (at procedure step  200 ).  
     [0239] 22. Systems for Executing Preferred Methods of the Invention.  
     [0240] An embodiment of computer, network and instrument resources for effecting the methods of the invention is shown in FIG. 18. In this embodiment, four computer servers are provided. First, a large database server  2002  stores all chemical structure, sample tracking and genomic, assay, quality control, and program status data. Further, this database server serves as the platform for a document management system. Second, a compute engine  2004  runs computational programs including RNA folding, oligonucleotide walking, and genomic searching. Third, a file server  2006  allows raw instrument output storage and sharing of robot instructions. Fourth, a groupware server  2008  enhances staff communication and process scheduling.  
     [0241] A redundant high-speed network system is provided between the main servers and the bridges  2026 ,  2028  and  2030 . These bridges provide reliable network access to the many workstations and instruments deployed for this process. The instruments selected to support this embodiment are all designed to sample directly from standard 96 well microtiter plates, and include an optical density reader  2016 , a combined liquid chromatography and mass spectroscopy instrument  2018 , a gel fluorescence and scintillation imaging system  2032  and  2042 , a capillary gel electrophoreses system  2022  and a real-time PCR system  2034 .  
     [0242] Most liquid handling is accomplished automatically using robots with individually controllable robotic pipetters  2038  and  2020  as well as a 96-well pipette system  2040  for duplicating plates. Windows NT or Macintosh workstations  2044 ,  2024 , and  2036  are deployed for instrument control, analysis and productivity support.  
     [0243] 23. Relational Database.  
     [0244] Data is stored in an appropriate database. For use with the methods of the invention, a relational database is preferred. FIG. 19 illustrates the data structure of a sample relational database. Various elements of data are segregated among linked storage elements of the database.  
     EXAMPLES  
     [0245] The following examples illustrate the invention and are not intended to limit the same. Those skilled in the art will recognize, or be able to ascertain through routine experimentation, numerous equivalents to the specific procedures, materials and devices described herein. Such equivalents are considered to be within the scope of the present invention.  
     Example 1  
     [0246] Selection of CD40 as a Target  
     [0247] Cell-cell interactions are a feature of a variety of biological processes. In the activation of the immune response, for example, one of the earliest detectable events in a normal inflammatory response is adhesion of leukocytes to the vascular endothelium, followed by migration of leukocytes out of the vasculature to the site of infection or injury. The adhesion of leukocytes to vascular endothelium is an obligate step in their migration out of the vasculature (for a review, see Albelda et al.,  FASEB J.,  1994, 8, 504). As is well known in the art, cell-cell interactions are also critical for propagation of both B-lymphocytes and T-lymphocytes resulting in enhanced humoral and cellular immune responses, respectively (for a reviews, see Makgoba et al.,  Immunol. Today,  1989, 10, 417; Janeway,  Sci. Amer.,  1993, 269, 72).  
     [0248] CD40 was first characterized as a receptor expressed on B-lymphocytes. It was later found that engagement of B-cell CD40 with CD40L expressed on activated T-cells is essential for T-cell dependent B-cell activation (i.e. proliferation, immunoglobulin secretion, and class switching) (for a review, see Gruss et al.  Leuk. Lymphoma,  1997, 24, 393). A full cDNA sequence for CD40 is available (GenBank accession number X60592, incorporated herein by reference as SEQ ID NO: 85).  
     [0249] As interest in CD40 mounted, it was subsequently revealed that functional CD40 is expressed on a variety of cell types other than B-cells, including macrophages, dendritic cells, thymic epithelial cells, Langerhans cells, and endothelial cells (Ibid.). These studies have led to the current belief that CD40 plays a much broader role in immune regulation by mediating interactions of T-cells with cell types other than B-cells. In support of this notion, it has been shown that stimulation of CD40 in macrophages and dendritic results is required for T-cell activation during antigen presentation (Id.). Recent evidence points to a role for CD40 in tissue inflammation as well. Production of the inflammatory mediators IL-12 and nitric oxide by macrophages has been shown to be CD40 dependent (Buhlmann et al.,  J. Clin. Immunol.,  1996, 16, 83). In endothelial cells, stimulation of CD40 by CD40L has been found to induce surface expression of E-selectin, ICAM-1, and VCAM-1, promoting adhesion of leukocytes to sites of inflammation (Buhlmann et al.,  J. Clin. Immunol,  1996, 16, 83; Gruss et al.,  Leuk Lymphoma,  1997, 24, 393). Finally, a number of reports have documented overexpression of CD40 in epithelial and hematopoietic tumors as well as tumor infiltrating endothelial cells, indicating that CD40 may play a role in tumor growth and/or angiogenesis as well (Gruss et al.,  Leuk Lymphoma,  1997, 24, 393-422; Kluth et al.  Cancer Res,  1997, 57, 891).  
     [0250] Due to the pivotal role that CD40 plays in humoral immunity, the potential exists that therapeutic strategies aimed at downregulating CD40 may provide a novel class of agents useful in treating a number of immune associated disorders, including but not limited to graft versus host disease, graft rejection, and autoimmune diseases such as multiple sclerosis, systemic lupus erythematosus, and certain forms of arthritis. Inhibitors of CD40 may also prove useful as an anti-inflammatory compound, and could therefore be useful as treatment for a variety of diseases with an inflammatory component such as asthma, rheumatoid arthritis, allograft rejections, inflammatory bowel disease, and various dermatological conditions, including psoriasis. Finally, as more is learned about the association between CD40 overexpression and tumor growth, inhibitors of CD40 may prove useful as anti-tumor agents as well.  
     [0251] Currently, there are no known therapeutic agents which effectively inhibit the synthesis of CD40. To date, strategies aimed at inhibiting CD40 function have involved the use of a variety of agents that disrupt CD40/CD40L binding. These include monoclonal antibodies directed against either CD40 or CD40L, soluble forms of CD40, and synthetic peptides derived from a second CD40 binding protein, A20. The use of neutralizing antibodies against CD40 and/or CD40L in animal models has provided evidence that inhibition of CD40 stimulation would have therapeutic benefit for GVHD, allograft rejection, rheumatoid arthritis, SLE, MS, and B-cell lymphoma (Buhlmann et al.,  J. Clin. Immunol,  1996, 16, 83). However, due to the expense, short half-life, and bioavailability problems associated with the use of large proteins as therapeutic agents, there is a long felt need for additional agents capable of effectively inhibiting CD40 function. Oligonucleotides compounds avoid many of the pitfalls of current agents used to block CD40/CD40L interactions and may therefore prove to be uniquely useful in a number of therapeutic applications.  
     Example 2  
     [0252] Generation of Virtual Oligonucleotides Targeted to CD40  
     [0253] The process of the invention was used to select oligonucleotides targeted to CD40, generating the list of oligonucleotide sequences with desired properties as shown in FIG. 22. From the assembled CD40 sequence, the process began with determining the desired oligonucleotide length to be eighteen nucleotides, as represented in step  2500 . All possible oligonucleotides of this length were generated by Oligo 5.0™, as represented in step  2504 . Desired thermodynamic properties were selected in step  2508 . The single parameter used was oligonucleotides of melting temperature less than or equal to 40° C. were discarded. In step  2512 , oligonucleotide melting temperatures were calculated by Oligo 5.0™. Oligonucleotide sequences possessing an undesirable score were discarded. It is believed that oligonucleotides with melting temperatures near or below physiological and cell culture temperatures will bind poorly to target sequences. All oligonucleotide sequences remaining were exported into a spreadsheet. In step  2516 , desired sequence properties are selected. These include discarding oligonucleotides with at least one stretch of four guanosines in a row and stretches of six of any other nucleotide in a row. In step  2520 , a spreadsheet macro removed all oligonucleotides containing the text string “GGGG.” In step  2524 , another spreadsheet macro removed all oligonucleotides containing the text strings “AAAAAA” or “CCCCCC” or “TTTTTT.” From the remaining oligonucleotide sequences, 84 sequences were selected manually with the criteria of having an uniform distribution of oligonucleotide sequences throughout the target sequence, as represented in step  2528 . These oligonucleotide sequences were then passed to the next step in the process, assigning actual oligonucleotide chemistries to the sequences.  
     Example 3  
     [0254] Input Files For Automated Oligonucleotide Synthesis Command File (.cmd File)  
     [0255] Table 2 is a command file for synthesis of an oligonucleotide having regions of 2′-O-(2-methoxyethyl) nucleosides and a central region of 2′-deoxy nucleosides each linked by phosphorothioate internucleotide linkages.  
                       TABLE 2                                      SOLID_SUPPORT_SKIP                         BEGIN                         Next_Sequence                         END                         INITIAL-WASH                         BEGIN                         Add ACN 300           Drain 10                         END                         LOOP-BEGIN           DEBLOCK                         BEGIN                         Prime TCA           Load Tray           Repeat 2                         Add TCA 150           Wait 10           Drain 8                         End_Repeat           Remove Tray                         Add TCA 125           Wait 10           Drain 8                         END                         WASH_AFTER_DEBLOCK                         BEGIN                         Repeat 3                         Add ACN 250 To_All           Drain 10                         End_Repeat                         END                         COUPLING                         BEGIN                         if class = DEOXY_THIOATE                         Nozzle wash &lt;act1&gt;           prime &lt;act1&gt;           prime &lt;seq&gt;           Add &lt;act1&gt; 70 + &lt;seq&gt; 70                         Wait 40                         Drain 5                         end-if                         if class = MOE_THIOATE                         Nozzle wash &lt;act1&gt;                         Prime &lt;act1&gt;           prime &lt;seq&gt;           Add &lt;act1&gt; 120 + &lt;seq&gt; 120           Wait 230           Drain 5                         End_if                         END                         WASH_AFTER_COUPLING                         BEGIN                         Add ACN 200 To_All           Drain 10                         END                         OXIDIZE                         BEGIN                         if class = DEOXY_THIOATE                         Add BEAU 180           Wait 40                         Drain 7                         end_if           if class = MOE_THIOATE           Add BEAU 200                         Wait 120           Drain 7                         end_if                         END                         CAP                         BEGIN                         Add CAP_B 80 + CAP_A 80           Wait 20           Drain 7                         END                         WASH_AFTER_CAP                         BEGIN                         Add ACN 150 To_All           Drain 5           Add ACN 250 To_All           Drain 11                         END                         BASE_COUNTER                         BEGIN                         Next_Sequence                         END                         LOOP_END           DEBLOCK_FINAL                         BEGIN                         Prime TCA           Load Tray           Repeat 2                         Add TCA 150 To_All           Wait 10           Drain 8                         End_Repeat           Remove Tray           Add TCA 125 To_All           Wait 10           Drain 10                         END                         FINAL_WASH                         BEGIN                         Repeat 4                         Add ACN 300 to_All           Drain_12                         End_Repeat                         END                         ENDALL                         BEGIN                         Wait 3                         END                      
 
     [0256] Sequence Files (.seq Files)  
     [0257] Table 3 is a .seq file for oligonucleotides having 2′-deoxy nucleosides linked by phosphorothioate internucleotide linkages.  
                   TABLE 3                       Identity of columns:             Syn #, Well, Scale, Nucleotide at particular           position  (identified using base identifier       followed by backbone identifier where “s” is       phosphorothioate). Note the columns wrap around       to next line when longer than one line.                                                                                            1   A01   200   As   Cs   Cs   As   Gs   Gs   As   Cs   Gs   Gs   Cs           2   A02   200   As   Cs   Gs   Gs   Cs   Gs   Gs   As   Cs   Cs   As       3   A03   200   As   Cs   Cs   As   As   Gs   Cs   As   Gs   As   Cs       4   A04   200   As   Gs   Gs   As   Gs   As   Cs   Cs   Cs   Cs   Gs       5   A05   200   As   Cs   Cs   Cs   Cs   Gs   As   Cs   Gs   As   As       6   A06   200   As   Cs   Gs   As   As   Cs   Gs   As   Cs   Ts   Gs       7   A07   200   As   Cs   Gs   As   Cs   Ts   Gs   Gs   Cs   Gs   As       8   A08   200   As   Cs   As   Gs   Gs   Ts   As   Gs   Gs   Ts   Cs       9   A09   200   As   Gs   Gs   Ts   Cs   Ts   Ts   Gs   Gs   Ts   Gs       10   A10   200   As   Cs   Ts   Cs   As   Cs   Cs   As   Cs   As   As       11   A11   200   As   Cs   Gs   As   Cs   As   As   Gs   As   As   As       12   A12   200   As   Cs   As   As   As   Cs   As   Cs   Gs   Gs   Ts       13   B01   200   As   As   Cs   As   Cs   Gs   Gs   Ts   Cs   Gs   Gs       14   B02   200   As   Cs   Ts   Cs   As   Cs   Ts   Gs   As   Cs   Gs       15   B03   200   As   Cs   Gs   Gs   As   As   Gs   Gs   As   As   Cs       16   B04   200   As   Ts   Cs   Ts   Gs   Ts   Gs   Gs   As   Cs   Cs       17   B05   200   As   Cs   As   Cs   Ts   Ts   Cs   Ts   Ts   Cs   Cs       18   B06   200   As   Cs   Ts   Cs   Ts   Cs   Gs   As   Cs   As   Cs       19   B07   200   As   As   As   Cs   Cs   Cs   Cs   As   Gs   Ts   Ts       20   B08   200   As   Ts   Gs   Ts   Cs   Cs   Cs   Cs   As   As   As       21   B09   200   As   Cs   Gs   Cs   Ts   Cs   Gs   Gs   Gs   As   Cs       22   B10   200   As   Gs   Cs   Cs   Gs   As   As   Gs   As   As   Gs       23   B11   200   As   Cs   As   Cs   As   Gs   Ts   As   Gs   As   Cs       24   B12   200   As   Cs   As   Cs   Ts   Cs   Ts   Gs   Gs   Ts   Ts       25   C01   200   As   Cs   Gs   As   Cs   Cs   As   Gs   As   As   As       26   C02   200   As   Gs   Ts   Ts   As   As   As   As   Gs   Gs   Gs       27   C03   200   As   Gs   Gs   Ts   Ts   Gs   Ts   Gs   As   Cs   Gs       28   C04   200   As   As   Ts   Gs   Ts   As   Cs   Cs   Ts   As   Cs       29   C05   200   As   Gs   Ts   Cs   As   Cs   Gs   Ts   Cs   Cs   Ts       30   C06   200   Cs   Ts   Gs   Gs   Cs   Gs   As   Cs   As   Gs   Gs       31   C07   200   Cs   Ts   Cs   Ts   Gs   Ts   Gs   Ts   Gs   As   Cs       32   C08   200   Cs   As   Gs   Gs   Ts   Cs   Gs   Ts   Cs   Ts   Ts       33   C09   200   Cs   Ts   Gs   Ts   Gs   Gs   Ts   As   Gs   As   Cs       34   C10   200   Cs   Ts   As   As   Cs   Gs   As   Ts   Gs   Ts   Cs       35   C11   200   Cs   Ts   Gs   Ts   Ts   Cs   Gs   As   Cs   As   Cs       36   C12   200   Cs   Ts   Gs   Gs   As   Cs   Cs   As   As   Cs   As       37   D01   200   Cs   Cs   Gs   Ts   Cs   Cs   Gs   Ts   Gs   Ts   Ts       38   D02   200   Cs   Ts   Gs   As   Cs   Ts   As   Cs   As   As   Cs       39   D03   200   Cs   As   As   Cs   As   Gs   As   Cs   As   Cs   Cs       40   D04   200   Cs   As   Gs   Gs   Gs   Gs   Ts   Cs   Cs   Ts   As       41   D05   200   Cs   Ts   Cs   Ts   As   Gs   Ts   Ts   As   As   As       42   D06   200   Cs   Ts   Gs   Cs   Ts   As   Gs   As   As   Gs   Gs       43   D07   200   Cs   Ts   Gs   As   As   As   Ts   Gs   Ts   As   Cs       44   D08   200   Cs   As   Cs   Cs   Cs   Gs   Ts   Ts   Ts   Gs   Ts       45   D09   200   Cs   Ts   Cs   Gs   As   Ts   As   Cs   Gs   Gs   Gs       46   D10   200   Gs   Gs   Ts   As   Gs   Gs   Ts   Cs   Ts   Ts   Gs       47   D11   200   Gs   As   Cs   Ts   Ts   Ts   Gs   Cs   Cs   Ts   Ts       48   D12   200   Gs   Ts   Gs   Gs   As   Gs   Ts   Cs   Ts   Ts   Ts       49   E01   200   Gs   Gs   As   Gs   Ts   Cs   Ts   Ts   Ts   Gs   Ts       50   E02   200   Gs   Gs   As   Cs   As   Cs   Ts   Cs   Ts   Cs   Gs       51   E03   200   Gs   As   Cs   As   Cs   As   Gs   Gs   As   Cs   Gs       52   E04   200   Gs   As   Gs   Ts   As   Cs   Gs   As   Gs   Cs   Gs       53   E05   200   Gs   As   Cs   Ts   As   Ts   Gs   Gs   Ts   As   Gs       54   E06   200   Gs   As   As   Gs   As   Gs   Gs   Ts   Ts   As   Cs       55   E07   200   Gs   As   Gs   Gs   Ts   Ts   As   Cs   As   Cs   As       56   E08   200   Gs   Ts   Ts   Gs   Ts   Cs   Cs   Gs   Ts   Cs   Cs       57   E09   200   Gs   As   Cs   Ts   Cs   Ts   Cs   Gs   Gs   Gs   As       58   E10   200   Gs   Ts   As   Gs   Gs   As   Gs   As   As   Cs   Cs       59   E11   200   Gs   Gs   Ts   Ts   Cs   Ts   Ts   Cs   Gs   Gs   Ts       60   E12   200   Gs   Ts   Gs   Gs   Gs   Gs   Ts   Ts   Cs   Gs   Ts       61   F01   200   Gs   Ts   Cs   As   Cs   Gs   Ts   Cs   Cs   Ts   Cs       62   F02   200   Gs   Ts   Cs   Cs   Ts   Cs   Cs   Ts   As   Cs   Cs       63   F03   200   Gs   Ts   Cs   Cs   Cs   Cs   As   Cs   Gs   Ts   Cs       64   F04   200   Ts   Cs   As   Cs   Cs   As   Gs   Cs   As   Gs   Cs       65   F05   200   Ts   As   Cs   Cs   As   As   Gs   Cs   As   Gs   As       66   F06   200   Ts   Cs   Cs   Ts   Gs   Ts   Cs   Ts   Ts   Ts   Gs       67   F07   200   Ts   Gs   Ts   Cs   Ts   Ts   Ts   Gs   As   Cs   Cs       68   F08   200   Ts   Gs   As   Cs   Cs   As   Cs   Ts   Cs   As   Cs       69   F09   200   Ts   Gs   As   Cs   Gs   Ts   Gs   Ts   Cs   Ts   Cs       70   F10   200   Ts   Cs   As   As   Gs   Ts   Gs   As   Cs   Ts   Ts       71   F11   200   Ts   Gs   Ts   Ts   Ts   As   Ts   Gs   As   Cs   Gs       72   F12   200   Ts   Ts   As   Ts   Gs   As   Cs   Gs   Cs   Ts   Gs       73   G01   200   Ts   Gs   As   Cs   Gs   Cs   Ts   Gs   Gs   Gs   Gs       74   G02   200   Ts   Cs   Gs   Ts   Cs   Ts   Ts   Cs   Cs   Cs   Gs       75   G03   200   Ts   Gs   Gs   Ts   As   Gs   As   Cs   Gs   Ts   Gs       76   G04   200   Ts   Ts   Cs   Ts   Ts   Cs   Cs   Gs   As   Cs   Cs       77   G05   200   Ts   Gs   Gs   Ts   As   Gs   As   Cs   Gs   Cs   Ts       78   G06   200   Ts   As   Gs   As   Cs   Gs   Cs   Ts   Cs   Gs   Gs       79   G07   200   Ts   Ts   Ts   Ts   As   Cs   As   Gs   Ts   Gs   Gs       80   G08   200   Ts   Gs   Gs   Gs   As   As   Cs   Cs   Ts   Gs   Ts       81   G09   200   Ts   Cs   Gs   Gs   Gs   As   Cs   Cs   As   Cs   Cs       82   G10   200   Ts   As   Gs   Gs   As   Cs   As   As   As   Cs   Gs       83   G11   200   Ts   Gs   Cs   Ts   As   Gs   As   As   Gs   Gs   As       84   G12   200   Ts   Cs   Ts   Gs   Ts   Cs   As   Cs   Ts   Cs   Cs                  
 
     [0258] Table 4 is a .seq file for oligonucleotides having regions of 2′-O-(2-methoxyethyl)nucleosides and a central region of 2′-deoxy nucleosides each linked by phosphorothioate internucleotide linkages.  
                   TABLE 4                       Identity of columns:             Syn #, Well, Scale, Nucleotide at particular position         (identified using base identifier followed by backbone       identifier where “s” is phosphorothioate and “moe”       indicated a 2′-O-(2-methoxyethyl) substituted nucleoside).       The columns wrap around to next line       when longer than one line.                                         1 A01 200 moeAs moeCs moeCs moeAs Gs Gs As Cs Gs Gs Cs Gs Gs As           moeCs moeCs moeAs moeG                2 A02 200 moeAs moeCs moeCs moeGs Cs Gs Gs As Cs Cs As Gs As Gs       moeTs moeGs moeGs moeA                3 A03 200 moeAs moeCs moeCs moeAs As Gs Cs As Gs As Cs Gs Gs As       moeGs moeAs moeCs moeG                4 A04 200 moeAs moeGs moeGs moeAs Gs As Cs Cs Cs Cs Gs As Cs Gs       moeAs moeAs moeCs moeG                5 A05 200 moeAs moeCs moeCs moeCs Cs Gs As Cs Gs As As Cs Gs As       moeCs moeTs moeGs moeG                6 A06 200 moeAs moeCs moeGs moeAs As Cs Gs As Cs Ts Gs Gs Cs Gs       moeAs moeCs moeAs moeG                7 A07 200 moeAs moeCs moeGs moeAs Cs Ts Gs Gs Cs Gs As Cs As Gs       moeGs moeTs moeAs moeG                8 A08 200 moeAs moeCs moeAs moeGs Gs Ts As Gs Gs Ts Cs Ts Ts Gs       moeGs moeTs moeGs moeG                9 A09 200 moeAs moeGs moeGs moels Cs Ts Ts Gs Gs Ts Gs Gs Gs Ts       moeGs moeAs moeCs moeG               10 A10 200 moeAs moeGs moeTs moeCs As Cs Gs As Cs As As Gs As As       moeAs moeCs moeAs moeC               11 A11 200 moeAs moeCs moeGs moeAs Cs As As Gs As As As Cs As Cs       moeGs moeGs moeTs moeC               12 A12 200 moeAs moeGs moeAs moeAs As Cs As Cs Gs Gs Ts Cs Gs Gs       moeTs moeCs moeCs moeT               13 B01 200 moeAs moeAs moeCs moeAs Cs Gs Gs Ts Cs Gs Gs Ts Cs Cs       moeTs moeGs moeTs moeC               14 B02 200 moeAs moeCs moeTs moeCs As Cs Ts Gs As Cs Gs Ts Gs Ts       moeCs moeTs moeCs moeA               15 B03 200 mocAs moeCs moeGs moeGs As As Gs Gs As As Cs Gs Cs Cs       moeAs moeCs moeTs moeT               16 B04 200 moeAs moeTs moeCs moeTs Gs Ts Gs Gs As Cs Cs Ts Ts Gs       moeTs moeCs moeTs moeC               17 B05 200 moeAs moeCs moeAs moeCs Ts Ts Cs Ts Ts Cs Cs Gs As Cs       moeCs moeGs moeTs moeG               18 B06 200 moeAs moeCs moeTs moeCs Ts Cs Gs As Cs As Cs As Gs Gs       moeAs moeCs moeGs moeT               19 B07 200 moeAs moeAs moeAs moeCs Cs Cs Cs As Gs Ts Ts Cs Gs Ts       moeCs moeTs moeAs moeA               20 B08 200 moeAs moeTs moeGs moeTs Cs Cs Cs Cs As As As Gs As Cs       moeTs moeAs moeTs moeG               21 B09 200 moeAs moeCs moeGs moeCs Ts Cs Gs Gs Gs As Cs Gs Gs Gs       moeTs moeCs moeAs moeG               22 B10 200 moeAs moeGs moeCs moeCs Gs As As Gs As As Gs As Gs Gs       moeTs moeTs moeAs moeC               23 B11 200 moeAs moeCs moeAs moeCs As Gs Ts As Gs As Cs Gs As As       moeAs moeGs moeCs moeT               24 B12 200 moeAs moeCs moeAs moeCs Ts Cs Ts Gs Gs Ts Ts Ts Cs Ts       moeGs moeGs moeAs moeC               25 C01 200 moeAs moeCs moeGs moeAs Cs Cs As Hs As As As Ts As Gs       moeTs moeTs moeTs moeT               26 C02 200 moeAs moeGs moeTs moeTs As As As As Gs Gs Gs Cs Ts Gs       moeCs moeTs moeAs moeG               27 C03 200 moeAs moeGs moeGs moeTs Ts Gs Ts Gs As Cs Gs As Cs Gs       moeAs moeGs moeGs moeT               28 C04 200 moeAs moeAs moeTs moeGs Ts As Cs Cs Ts As Cs Gs Gs Ts       moeTs moeGs moeGs moeC               29 CO5 200 moeAs moeGs moeTs moeCs As Cs Gs Ts Cs Cs Ts Cs Ts Cs       moeTs moeGs moeTs moeC               30 C06 200 moeCs moeTs moeGs moeGs Cs Gs As Cs As Gs Gs Ts As Gs       moeGs moeTs moeCs moeT               31 C07 200 moeCs moeTs moeGs moeTs Gs Ts Gs Ts Gs As Cs Gs Gs Ts       moeGs moeGs moeTs moeC               32 C08 200 moeCs moeAs moeGs moeGs Ts Cs Gs Ts Cs Ts Ts Cs Cs Cs       moeGs moeTs moeGs moeG               33 C09 200 moeCs moeTs moeGs moeTs Gs Gs Ts As Gs As Cs Gs Ts Gs       moeGs moeAs moeCs moeA               34 C10 200 moeCs moeTs moeAs moeAs Cs Gs As Ts Gs Ts Cs Cs Cs Cs       moeAs rnoeAs moeAs moeG               35 C11 200 moeCs moeTs moeGs moeTs Ts Cs Gs As Cs As Cs Ts Cs Ts       moeGs moeGs moeTs moeT               36 C12 200 moeCs moeTs moeGs moeGs As Cs Cs As As Cs As Cs Gs Ts       moeTs macGs moeTs moeC               37 D01 200 moeCs moeCs moeGs moeTs Cs Cs Gs Ts Gs Ts Ts Ts Gs Ts       moeTs moeCs moeTs moeG               38 D02 200 moeCs moeTs moeGs mocAs Cs Ts As Cs As As Cs As Gs As       moeCs moeAs moeCs moeC               39 D03 200 moeCs moeAs moeAs moeCs As Gs As Cs As Cs Cs As Gs Gs       moeGs moeGs moels moeC               40 D04 200 moeCs moeAs moeGs moeGs Gs Gs Ts Cs Cs Ts As Gs Cs Cs       moeGs moeAs moeCs moeT               41 D05 200 moeCs moeTs moeCs moeTs As Gs Ts Ts As As As As Gs Gs       moeGs moeCs moeTs moeG               42 D06 200 moeCs moeTs moeGs moeCs Ts As Gs As As Gs Gs As Cs Cs       moeGs moeAs moeGs moeG               43 D07 200 moeCs moeTs moeGs moeAs As As Ts Gs Ts As Cs Cs Ts As       moeCs moeGs moeGs moeT               44 D08 200 moeCs moeAs moeCs moeCs Cs Gs Ts Ts Ts Cs Ts Cs Cs Gs       moeTs moeCs moeAs moeA               45 D09 200 moeCs moeTs moeCs moeGs As Ts As Cs Gs Gs Gs Ts Cs As       moeGs moeTs moeCs moeA               46 D10 200 moeGs moeGs moeTs moeAs Gs Gs Ts Cs Ts Ts Gs Gs Ts Gs       moeGs moeGs moeTs moeG               47 D11 200 moeGs moeAs moeCs moeTs Ts Ts Gs Cs Cs Ts Ts As Cs Gs       moeGs moeAs moeAs moeG               48 D12 200 moeGs moeTs moeGs moeGs As Gs Ts Cs Ts Ts Ts Gs Ts Cs       moeTs moeGs moeTs moeG               49 E01 200 moeGs moeGs moeAs moeGs Ts Cs Ts Ts Ts Gs Ts Cs Ts Gs       moeTs moeGs moeGs moeT               50 E02 200 moeGs moeGs moeAs moeCs As Cs Ts Cs Ts Cs Gs As Cs As       moeCs moeAs moeGs moeG               51 E03 200 moeGs moeAs moeCs moeAs Cs As Gs Gs As Cs Gs Ts Gs Gs       moeCs moeGs moeAs moeG               52 E04 200 moeGs moeAs moeGs moeTs As Cs Gs As Gs Cs Gs Gs Gs Cs       moeCs moeGs moeAs moeA               53 E05 200 moeGs moeAs moeCs moeTs As Ts Gs Gs Ts As Gs As Cs Gs       moeCs moeTs moeCs moeG               54 E06 200 moeGs moeAs moeAs moeGs As Gs Gs Ts Ts As Cs As Cs As       moeGs moeTs moeAs moeG               55 E07 200 moeGs moeAs moeGs moeGs Ts Ts As Cs As Cs As Gs Ts As       moeGs moeAs moeCs moeG               56 E08 200 moeGs moeTs moeTs moeGs Ts Cs Cs Gs Ts Cs Cs Gs Ts Gs       moeTs moels moeTs moeG               57 E09 200 moeGs moeAs moeCs moeTs Cs Ts Cs Gs Gs Gs As Cs Cs As       moeCs moeCs moeAs moeC               58 E10 200 moeGs moeTs moeAs moeGs Gs As Gs As As Cs Cs As Cs Gs       moeAs moeCs moeCs moeA               59 E11 200 moeGs moeGs moeTs moels Cs Ts Ts Cs Gs Gs Ts Ts Gs Gs       moeTs moeTs moeAs moeT               60 E12 200 moeGs moeTs moeGs moeGs Gs Gs Ts Ts Cs Gs Ts Cs Cs Ts       moeTs moeGs moeGs moeG               61 F01 200 moeGs moeTs moeCs moeAs Cs Gs Ts Cs Cs Ts Cs Ts Gs As       moeAs moeAs moeTs moeG               62 F02 200 moeGs moeTs moeCs moeCs Ts Cs Cs Ts As Cs Cs Gs Ts Ts       moeTs moeCs moeTs moeC               63 F03 200 moeGs moeTs moeCs moeCs Cs Cs As Cs Gs Ts Cs Cs Gs Ts       moeCs moeTs moeTs moeC               64 F04 200 moeTs moeCs moeAs moeCs Cs As Gs Gs As Cs Gs Gs Cs Gs       moeGs moeAs moeCs moeC               65 F05 200 moeTs mocAs moeCs moeCs As As Gs Cs As Gs As Cs Gs Gs       moeAs moeGs moeAs moeC               66 F06 200 moeTs moeCs moeCs moeTs Gs Ts Cs Ts Ts Ts Gs As Cs Cs       moeAs moeCs moeTs moeC               67 F07 200 moeTs moeGs moeTs moeCs Ts Ts Ts Gs As Cs Cs As Gs Ts       moeCs moeAs moeCs moeT               68 F08 200 moeTs moeGs moeAs moeCs Cs As Cs Ts Cs As Cs Ts Gs As       moeCs moeGs moeTs moeG               69 F09 200 moeTs moeGs moeAs moeCs Gs Ts Gs Ts Cs Ts Cs As As Gs       moeTs moeGs moeAs moeC               70 F10 200 moeTs moeCs moeAs moeAs Gs Ts Gs As Cs Ts Ts Ts Gs Cs       moeCs moeTs moeTs moeA               71 F11 200 moeTs moeGs moeTs moeTs Ts As Ts Cs As Cs Gs Cs Ts Gs       moeGs moeGs moeGs moeT               72 F12 200 moeTs moeTs moeAs moeTs Gs As Cs Gs Cs Ts Gs Gs Gs Gs       moeTs moeTs moeGs moeG               73 G01 200 moeTs moeGs moeAs moeCs Gs Cs Ts Gs Gs Gs Gs Ts Ts Gs       moeGs moeAs moeTs moeC               74 G02 200 moeTs moeCs moeGs moeTs Cs Ts Ts Cs Cs Cs Gs Ts Gs Gs       moeAs moeGs moeTs moeC               75 G03 200 moeTs moeGs moeGs moeTs As Gs As Cs Gs Ts Gs Gs As Cs       moeAs moeCs moeTs moeT               76 G04 200 moeTs moeTs moeCs moeTs Ts Cs Cs Gs As Cs Cs Gs Ts Gs       moeAs moeCs moeAs moeT               77 G05 200 moeTs moeCs moeCs moeTs As Gs As Cs Gs Cs Ts Cs Gs Gs       moeGs moeAs moeCs moeG               78 G06 200 moeTs moeAs moeGs moeAs Cs Gs Cs Ts Cs Gs Gs Gs As Cs       moeGs moeGs moeGs moeT               79 G07 200 moeTs moeTs moeTs moeTs As Cs As Gs Ts Gs Gs Gs As As       moeCs moeCs moeTs moeG               80 G08 200 moeTs moeGs moeGs moeGs As As Cs Cs Ts Gs Ts Ts Cs Gs       moeAs moeCs moeAs moeC               81 G09 200 moeTs moeCs moeGs moeGs Gs As Cs Cs As Cs Cs As Cs Ts       moeAs moeGs moeGs moeG               82 G10 200 moeTs moeAs moeGs moeGs As Cs As As As Cs Gs Gs Ts As       moeGs moeGs moeAs moeG               83 G11 200 moeTs moeGs moeCs moeTs As Gs As As Gs Gs As Cs Cs Gs       moeAs moeGs moeGs moeT               84 G12 200 moeTs moeCs moeTs moeGs Ts Cs As Cs Ts Cs Cs Gs As Cs       moeGs moeTs moeGs moeG                  
 
     [0259] Reagent File (.tab File)  
     [0260] Table 5 is a .tab file for reagents necessary for synthesizing an oligonucleotides having both 2′-O-(2-methoxyethyl)nucleosides and 2′-deoxy nucleosides located therein.  
               TABLE 5                       Identity of columns: GroupName, Bottle ID, ReagentName,       FlowRate, Concentration. Wherein reagent name is       identified using base identifier, “moe” indicated       a 2′-O-(2-methoxyethyl) substituted nucleoside       and “cpg” indicates a control pore glass solid support       medium. The columns wrap around to next line when longer       than one line.                                    SUPPORT                         BEGIN                                             0   moeG   moeG   cpg   100   1           0   moe5meC   moe5meC   cpg   100   1           0   moeA   moeA   cpg   100   1           0   moeT   moeT   cpg   100   1           END                 DEBLOCK                                         BEGIN                           70   TCA   TCA   100   1           END                 WASH                                         BEGIN                           65   ACN   ACN   190   1           END                 OXIDIZERS                                         BEGIN                           68   BEAU   BEAUCAGE   320   1           END                 CAPPING                         BEGIN                                     66   CAP_BCAP_B   220   1                                         67   CAP_A   CAP_A   230   1           END                 DEOXY THIOATE                         BEGIN                                         31,32   Gs   deoxyG   270   1                                     39,40   5meCs 5methyldeoxyC   270   1                                         37,38   As   deoxyA   270   1                                         29,30   Ts   deoxyT   270   1           END                 MOE-THIOATE                         BEGIN                                     15,16   moeGs methoxyethoxyG   240   1                                         23,24   moe5meCs   methoxyethoxyC   240   1           21,22   moeAs   methoxyethoxyA   240   1           13,14   moeTs   methoxyethoxyT   240   1           END                 ACTIVATORS                         BEGIN           5,6,7,8 SET   s-ethyl-tet  280                         Activates           DEOXY_THIOATE           MOE_THIOATE                         END                      
 
     Example 4  
     [0261] Synthesis of Nucleoside Phosphoramidites  
     [0262] The following compounds, including amidites and their intermediates were prepared as described in U.S. Pat. No. 6,426,220 and published PCT WO 02/36743; 5′-O-Dimethoxytrityl-thymidine intermediate for 5-methyl dC amidite, 5′-O-Dimethoxytrityl-2′-deoxy-5-methylcytidine intermediate for 5-methyl-dC amidite, 5′-O-Dimethoxytrityl-2′-deoxy-N4-benzoyl-5-methylcytidine penultimate intermediate for 5-methyl dC amidite, [5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-deoxy-N 4 -benzoyl-5-methylcytidin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite (5-methyl dC amidite), 2′-Fluorodeoxyadenosine, 2′-Fluorodeoxyguanosine, 2′-Fluorouridine, 2′-Fluorodeoxycytidine, 2′-O-(2-Methoxyethyl) modified amidites, 2′-O-(2-methoxyethyl)-5-methyluridine intermediate, 5′-O-DMT-2′-O-(2-methoxyethyl)-5-methyluridine penultimate intermediate, [5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-5-methyluridin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite (MOE T amidite), 5′-O-Dimethoxytrityl-2′-O-(2-methoxyethyl)-5-methylcytidine intermediate, 5′-O-dimethoxytrityl-2′-O-(2-methoxyethyl)-N 4 -benzoyl-5-methyl-cytidine penultimate intermediate, [5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N4-benzoyl-5-methylcytidin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite (MOE 5-Me-C amidite), [5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N 6 -benzoyladenosin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite (MOE A amdite), [5′-O-(4,4′-Dimethoxytriphenylmethyl)-2′-O-(2-methoxyethyl)-N 4 -isobutyrylguanosin-3′-O-yl]-2-cyanoethyl-N,N-diisopropylphosphoramidite (MOE G amidite), 2′-O-(Aminooxyethyl) nucleoside amidites and 2′-O-(dimethylaminooxyethyl) nucleoside amidites, 2′-(Dimethylaminooxyethoxy) nucleoside amidites, 5′-O-tert-Butyldiphenylsilyl-O 2 -2′-anhydro-5-methyluridine, 5′-O-tert-Butyldiphenylsilyl-2′-O-(2-hydroxyethyl)-5-methyluridine, 2′-O-([2-phthalimidoxy)ethyl]-5′-t-butyldiphenylsilyl-5-methyluridine, 5′-O-tert-butyldiphenylsilyl-2′-O-[(2-formadoximinooxy)ethyl]-5-methyluridine, 5′-O-tert-Butyldiphenylsilyl-2′-O-[N,N dimethylaminooxyethyl]-5-methyluridine, 2′-O-(dimethylaminooxyethyl)-5-methyluridine, 5′-O-DMT-2′-O-(dimethylaminooxyethyl)-5-methyluridine, 5′-O-DMT-2′-O-(2-N,N-dimethylaminooxyethyl)-5-methyluridine-3′-[(2-cyanoethyl)-N,N-diisopropylphosphoramidite], 2′-(Aminooxyethoxy) nucleoside amidites, N2-isobutyryl-6-O-diphenylcarbamoyl-2′-O-(2-ethylacetyl)-5′-O-(4,4′-dimethoxytrityl)guanosine-3′-[(2-cyanoethyl)-N,N-diisopropylphosphoramidite], 2′-dimethylaminoethoxyethoxy (2′-DMAEOE) nucleoside amidites, 2′-O-[2(2-N,N-dimethylaminoethoxy)ethyl]-5-methyl uridine, 5′-O-dimethoxytrityl-2′-O-[2(2-N,N-dimethylaminoethoxy)-ethyl)]-5-methyl uridine and 5′-O-Dimethoxytrityl-2′-O-[2(2-N,N-dimethylaminoethoxy)-ethyl)]-5-methyl uridine-3′-O-(cyanoethyl-N,N-diisopropyl)phosphoramidite.  
     Example 5  
     [0263] Oligonucleotide and Oligonucleoside Synthesis  
     [0264] The antisense compounds used in accordance with this invention may be conveniently and routinely made through the well-known technique of solid phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems (Foster City, Calif.). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is well known to use similar techniques to prepare oligonucleotides such as the phosphorothioates and alkylated derivatives.  
     [0265] Oligonucleotides: Unsubstituted and substituted phosphodiester (P═O) oligonucleotides are synthesized on an automated DNA synthesizer (Applied Biosystems model 394) using standard phosphoramidite chemistry with oxidation by iodine.  
     [0266] Phosphorothioates (P═S) are synthesized similar to phosphodiester oligonucleotides with the following exceptions: thiation was effected by utilizing a 10% w/v solution of 3,H-1,2-benzodithiole-3-one 1,1-dioxide in acetonitrile for the oxidation of the phosphite linkages. The thiation reaction step time was increased to 180 sec and preceded by the normal capping step. After cleavage from the CPG column and deblocking in concentrated ammonium hydroxide at 55° C. (12-16 hr), the oligonucleotides were recovered by precipitating with &gt;3 volumes of ethanol from a 1 M NH4OAc solution. Phosphinate oligonucleotides are prepared as described in U.S. Pat. No. 5,508,270, herein incorporated by reference.  
     [0267] Alkyl phosphonate oligonucleotides are prepared as described in U.S. Pat. No. 4,469,863, herein incorporated by reference.  
     [0268] 3+-Deoxy-3′-methylene phosphonate oligonucleotides are prepared as described in U.S. Pat. Nos. 5,610,289 or 5,625,050, herein incorporated by reference.  
     [0269] Phosphoramidite oligonucleotides are prepared as described in U.S. Pat. No. 5,256,775 or U.S. Pat. No. 5,366,878, herein incorporated by reference.  
     [0270] Alkylphosphonothioate oligonucleotides are prepared as described in published PCT applications PCT/US94/00902 and PCT/US93/06976 (published as WO 94/17093 and WO 94/02499, respectively), herein incorporated by reference.  
     [0271] 3′-Deoxy-3′-amino phosphoramidate oligonucleotides are prepared as described in U.S. Pat. No. 5,476,925, herein incorporated by reference.  
     [0272] Phosphotriester oligonucleotides are prepared as described in U.S. Pat. No. 5,023,243, herein incorporated by reference.  
     [0273] Borano phosphate oligonucleotides are prepared as described in U.S. Pat. Nos. 5,130,302 and 5,177,198, both herein incorporated by reference.  
     [0274] Oligonucleosides: Methylenemethylimino linked oligonucleosides, also identified as MMI linked oligonucleosides, methylenedi-methyl-hydrazo linked oligonucleosides, also identified as MDH linked oligonucleosides, and methylenecarbonylamino linked oligonucleosides, also identified as amide-3 linked oligonucleosides, and methyleneaminocarbonyl linked oligo-nucleosides, also identified as amide-4 linked oligo-nucleo-sides, as well as mixed backbone compounds having, for instance, alternating MMI and P═O or P═S linkages are prepared as described in U.S. Pat. Nos. 5,378,825, 5,386,023, 5,489,677, 5,602,240 and 5,610,289, all of which are herein incorporated by reference.  
     [0275] Formacetal and thioformacetal linked oligo-nucleo-sides are prepared as described in U.S. Pat. Nos. 5,264,562 and 5,264,564, herein incorporated by reference.  
     [0276] Ethylene oxide linked oligonucleosides are prepared as described in U.S. Pat. No. 5,223,618, herein incorporated by reference.  
     Example 6  
     [0277] Oligonucleotide Isolation  
     [0278] After cleavage from the controlled pore glass solid support and deblocking in concentrated ammonium hydroxide at 55° C. for 12-16 hours, the oligonucleotides or oligonucleosides are recovered by precipitation out of 1 M NH4OAc with &gt;3 volumes of ethanol. Synthesized oligonucleotides were analyzed by electrospray mass spectroscopy (molecular weight determination) and by capillary gel electrophoresis and judged to be at least 70% full length material. The relative amounts of phosphorothioate and phosphodiester linkages obtained in the synthesis was determined by the ratio of correct molecular weight relative to the −16 amu product (±32±48). For some studies oligonucleotides were purified by HPLC, as described by Chiang et al.,  J. Biol. Chem.  1991, 266, 18162-18171. Results obtained with HPLC-purified material were similar to those obtained with non-HPLC purified material.  
     Example 7  
     [0279] Chimeric Oligonucleotide Synthesis  
     [0280] Chimeric oligonucleotides, oligonucleosides or mixed oligonucleotides/oligonucleosides of the invention can be of several different types. These include a first type wherein the “gap” segment of linked nucleosides is positioned between 5′ and 3′ “wing” segments of linked nucleosides and a second “open end” type wherein the “gap” segment is located at either the 3′ or the 5′ terminus of the oligomeric compound. Oligonucleotides of the first type are also known in the art as “gapmers” or gapped oligonucleotides. Oligonucleotides of the second type are also known in the art as “hemimers” or “wingmers.” 
     [0281] A. [2′-O-Me]—[2′-deoxy]—[2′-O-Me] Chimeric Phosphorothioate Oligonucleotides  
     [0282] Chimeric oligonucleotides having 2′-O-alkyl phosphorothioate and 2′-deoxy phosphorothioate oligonucleotide segments are synthesized using 2′-deoxy-5′-dimethoxytrityl-3′-O-phosphoramidites for the DNA portion and 5′-dimethoxytrityl-2′-O-methyl-3′-O-phosphoramidites for 5′ and 3′ wings. The standard synthesis cycle is modified by increasing the wait step after the delivery of tetrazole and base to 600 s repeated four times for DNA and twice for 2′-O-methyl. The fully protected oligonucleotide was cleaved from the support and the phosphate group is deprotected in 3:1 Ammonia/Ethanol at room temperature overnight then lyophilized to dryness. Treatment in methanolic ammonia for 24 hrs at room temperature is done to deprotect all bases and the samples are again lyophilized to dryness.  
     [0283] B. [2′-O-(2-Methoxyethyl)]—[2′-deoxy]—[2′-O-(2-Methoxyethyl)] Chimeric Phosphorothioate Oligonucleotides  
     [0284] 2′-O-(2-methoxyethyl)]—[2′-deoxy]—[-2′-O-(2-methoxyethyl)] chimeric phosphorothioate oligonucleotides are prepared as per the procedure above for the 2′-O-methyl chimeric oligonucleotide, with the substitution of 2′-O-(2-methoxyethyl) amidites for the 2′-O-methyl amidites.  
     [0285] C. [2′-O-(2-Methoxyethyl)Phosphodiester]—[2′-deoxy Phosphoro-thioate]—[2′-O-(2-Methoxyethyl) Phosphodiester] Chimeric Oligo-Nucleotide  
     [0286] 2′-O-(2-methoxyethyl phosphodiester]—[2′-deoxy phosphorothioate]—[2′-O-(2-methoxyethyl) phosphodiester] chimeric oligonucleotides are prepared as per the above procedure for the 2′-O-methyl chimeric oligonucleotide with the substitution of 2′-O-(2-methoxyethyl) amidites for the 2′-O-methyl amidites in the wing portions. Sulfurization utilizing 3,H-1,2 benzodithiole-3-one 1,1 dioxide (Beaucage Reagent) is used to generate the phosphorothioate internucleotide linkages within the wing portions of the chimeric structures. Oxidization with iodine is used to generate the phosphodiester inter-nucleotide linkages for the center gap.  
     [0287] Other chimeric oligonucleotides, chimeric oligonucleosides and mixed chimeric oligonucleotides/oligonucleosides are synthesized according to U.S. Pat. No. 5,623,065, which is incorporated herein by reference in its entirety.  
     Example 8  
     [0288] Oligonucleotide Analysis  
     [0289] Oligonucleotides are analyzed by mass spectrometry as follows: for a typical oligonucleotide, a 0.01 OD aliquot dissolved in 10 uL of water is mixed with 90 uL of a 1:1 mixture of acetonitrile and water containing 20 mM imidazole and 20 mM piperidine (Greig M. J., Griffey R. H.:  Rapid Commun. Mass Spec.,  9:97-102, 1995). The sample is transferred to a 96 or 384-well plate, and each well on the plate is sampled systematically using an Agilent 1100 liquid handler. Other types of robotic liquid handlers such as a Leap Pal or Gilson 215 could be used to introduce sample to the mass spectrometer under computer control. The sample is infused to an Agilent MSD VX quadrupole mass spectrometer at a rate of 3 uL/min. The electrospray ionization is produced with 60 psi of nitrogen gas and a 4 kV potential between the source needle and the inlet capillary. The capillary is heated to 250° C. to effect desolvation of the ions. Typically, a total of 32 accumulations are averaged over a mass/charge range of 500-1500 m/z. The resulting data is saved into a file containing the sample ID information, and deconvoluted using the Agilent algorithm to calculate the neutral masses and abundances of the oligonucleotides and associated impurities. In a preferred embodiment, the neutral masses and abundances of compounds present in the sample obtained from the ESI-MS spectrum are written to a relational database. A logical algorithm then compares the measured mass of the oligonucleotide to the mass calculated from the base sequence and expected chemical structure of the oligonucleotide stored in a relational database. If the calculated and observed masses for the most abundant species agree within ±1.5 Da, the oligonucleotide is deemed to have “passed”. If the measured mass of the most abundant species differs by more than 1.5 Da, or the integrated ion abundance is &lt;50% of the sample, the oligonucleotide “fails” and a new synthesis is requested.  
     [0290] Oligonucleotides that pass the ESI-MS analysis are transferred to 96-well master plates for storage at 10 mM concentrations in aqueous solution using multichannel robotics, such as a Beckman FX or Packard MultiProbe. The plates are given sequential identifying bar codes and this information on sample location is stored for later retrieval in a relational database.  
     Example 9  
     [0291] PNA Synthesis  
     [0292] Peptide nucleic acids (PNAs) are prepared in accordance with any of the various procedures referred to in Peptide Nucleic Acids (PNA): Synthesis, Properties and Potential Applications,  Bioorganic  &amp;  Medicinal Chemistry,  1996, 4, 5. They may also be prepared in accordance with U.S. Pat. Nos. 5,539,082; 5,700,922, and 5,719,262, each of which is incorporated herein by reference in its entirety.  
     Example 10  
     [0293] RNA Synthesis  
     [0294] In general, RNA synthesis chemistry is based on the selective incorporation of various protecting groups at strategic intermediary reactions. Although one of ordinary skill in the art will understand the use of protecting groups in organic synthesis, a useful class of protecting groups includes silyl ethers. In particular bulky silyl ethers are used to protect the 5′-hydroxyl in combination with an acid-labile orthoester protecting group on the 2′-hydroxyl. This set of protecting groups is then used with standard solid-phase synthesis technology. It is important to lastly remove the acid labile orthoester protecting group after all other synthetic steps. Moreover, the early use of the silyl protecting groups during synthesis ensures facile removal when desired, without undesired deprotection of 2′-hydroxyl.  
     [0295] Following this procedure for the sequential protection of the 5′-hydroxyl in combination with protection of the 2′-hydroxyl by protecting groups that are differentially removed and are differentially chemically labile, RNA oligonucleotides were synthesized.  
     [0296] RNA oligonucleotides are synthesized in a stepwise fashion. Each nucleotide is added sequentially (3′- to 5′-direction) to a solid support-bound oligonucleotide. The first nucleoside at the 3′-end of the chain is covalently attached to a solid support. The nucleotide precursor, a ribonucleoside phosphoramidite, and activator are added, coupling the second base onto the 5′-end of the first nucleoside. The support is washed and any unreacted 5′-hydroxyl groups are capped with acetic anhydride to yield 5′-acetyl moieties. The linkage is then oxidized to the more stable and ultimately desired P(V) linkage. At the end of the nucleotide addition cycle, the 5′-silyl group is cleaved with fluoride. The cycle is repeated for each subsequent nucleotide.  
     [0297] Following synthesis, the methyl protecting groups on the phosphates are cleaved in 30 minutes utilizing 1 M disodium-2-carbamoyl-2-cyanoethylene-1,1-dithiolate trihydrate (S 2 Na 2 ) in DMF. The deprotection solution is washed from the solid support-bound oligonucleotide using water. The support is then treated with 40% methylamine in water for 10 minutes at 55° C. This releases the RNA oligonucleotides into solution, deprotects the exocyclic amines, and modifies the 2′-groups. The oligonucleotides can be analyzed by anion exchange HPLC at this stage.  
     [0298] The 2′-orthoester groups are the last protecting groups to be removed. The ethylene glycol monoacetate orthoester protecting group developed by Dharmacon Research, Inc. (Lafayette, Colo.), is one example of a useful orthoester protecting group which, has the following important properties. It is stable to the conditions of nucleoside phosphoramidite synthesis and oligonucleotide synthesis. However, after oligonucleotide synthesis the oligonucleotide is treated with methylamine which not only cleaves the oligonucleotide from the solid support but also removes the acetyl groups from the orthoesters. The resulting 2-ethyl-hydroxyl substituents on the orthoester are less electron withdrawing than the acetylated precursor. As a result, the modified orthoester becomes more labile to acid-catalyzed hydrolysis. Specifically, the rate of cleavage is approximately 10 times faster after the acetyl groups are removed. Therefore, this orthoester possesses sufficient stability in order to be compatible with oligonucleotide synthesis and yet, when subsequently modified, permits deprotection to be carried out under relatively mild aqueous conditions compatible with the final RNA oligonucleotide product.  
     [0299] Additionally, methods of RNA synthesis are well known in the art (Scaringe, S. A. Ph.D. Thesis, University of Colorado, 1996; Scaringe, S. A., et al.,  J. Am. Chem. Soc.,  1998, 120, 11820-11821; Matteucci, M. D. and Caruthers, M. H.  J. Am. Chem. Soc.,  1981, 103, 3185-3191; Beaucage, S. L. and Caruthers, M. H.  Tetrahedron Lett.,  1981, 22, 1859-1862; Dahl, B. J., et al.,  Acta Chem. Scand,.  1990, 44, 639-641; Reddy, M. P., et al.,  Tetrahedrom Lett.,  1994, 25, 4311-4314; Wincott, F. et al.,  Nucleic Acids Res.,  1995, 23, 2677-2684; Griffin, B. E., et al.,  Tetrahedron,  1967, 23, 2301-2313; Griffin, B. E., et al.,  Tetrahedron,  1967, 23, 2315-2331).  
     [0300] RNA antisense compounds (RNA oligonucleotides) of the present invention can be synthesized by the methods herein or purchased from Dharmacon Research, Inc (Lafayette, Colo.). Once synthesized, complementary RNA antisense compounds can then be annealed by methods known in the art to form double stranded (duplexed) antisense compounds. For example, duplexes can be formed by combining 30 μl of each of the complementary strands of RNA oligonucleotides (50 uM RNA oligonucleotide solution) and 15 μl of 5X annealing buffer (100 mM potassium acetate, 30 mM HEPES-KOH pH 7.4, 2 mM magnesium acetate) followed by heating for 1 minute at 90° C., then 1 hour at 37° C. The resulting duplexed antisense compounds can be used in kits, assays, screens, or other methods to investigate the role of a target nucleic acid.  
     Example 11  
     [0301] Output Oligonucleotides from Automated Oligonucleotide Synthesis  
     [0302] Using the .seq files, the .cmd files and .tab file of Example 3, oligonucleotides were prepared as per the protocol of the 96 well format of Example 5. The oligonucleotides were prepared utilizing phosphorothioate chemistry to give in one instance a first library of phosphorothioate oligodeoxynucleotides. The oligonucleotides were prepared in a second instance as a second library of hybrid oligonucleotides having phosphorothioate backbones with a first and third “wing” region of 2′-O-(2-methoxyethyl)nucleotides on either side of a center gap region of 2′-deoxy nucleotides. The two libraries contained the same set of oligonucleotide sequences. Thus the two libraries are redundant with respect to sequence but are unique with respect to the combination of sequence and chemistry. Because the sequences of the second library of compounds is the same as the first (however the chemistry is different), for brevity sake, the second library is not shown.  
     [0303] For illustrative purposes Tables 6-a and 6-b show the sequences of an initial first library, i.e., a library of phosphorothioate oligonucleotides targeted to a CD40 target. The compounds of Table 6-a shows the members of this library listed in compliance with the established rule for listing SEQ ID NO:, i.e., in numerical SEQ ID NO: order.  
                   TABLE 6-a                          Sequences of Oligonucleotides           Targeted to CD40 by SEQ ID NO.:                             NUCLEOBASE SEQUENCE   SEQ ID NO.                                             CCAGGCGGCAGGACCACT   1                           GACCAGGCGGCAGGACCA   2                       AGGTGAGACCAGGCGGCA   3                       CAGAGGCAGACGAACCAT   4                       GCAGAGGCAGACGAACCA   5                       GCAAGCAGCCCCAGAGGA   6                       GGTCAGCAAGCAGCCCCA   7                       GACAGCGGTCAGCAAGCA   8                       GATGGACAGCGGTCAGCA   9                       TCTGGATGGACAGCGGTC   10                       GGTGGTTCTGGATGGACA   11                       GTGGGTGGTTCTGGATGG   12                       GCAGTGGGTGGTTCTGGA   13                       CACAAAGAACAGCACTGA   14                       CTGGCACAAAGAACAGCA   15                       TCCTGGCTGGCACAAAGA   16                       CTGTCCTGGCTGGCACAA   17                       CTCACCAGTTTCTGTCCT   18                       TCACTCACCAGTTTCTGT   19                       GTGCAGTCACTCACCAGT   20                       ACTCTGTGCAGTCACTCA   21                       CAGTGAACTCTGTGCAGT   22                       ATTCCGTTTCAGTGAACT   23                       GAAGGCATTCCGTTTCAG   24                       TTCACCGCAAGGAAGGCA   25                       CTCTGTTCCAGGTGTCTA   26                       CTGGTGGCAGTGTGTCTC   27                       TGGGGTCGCAGTATTTGT   28                       GGTTGGGGTCGCAGTATT   29                       CTAGGTTGGGGTCGCAGT   30                       GGTGCCCTTCTGCTGGAC   31                       CTGAGGTGCCCTTCTGCT   32                       GTGTCTGTTTCTGAGGTG   33                       TGGTGTCTGTTTCTGAGG   34                       ACAGGTGCAGATGGTGTC   35                       TTCACAGGTGCAGATGGT   36                       GTGCCAGCCTTCTTCACA   37                       TACAGTGCCAGCCTTCTT   38                       GGACACAGCTCTCACAGG   39                       TGCAGGACACAGCTCTCA   40                       GAGCGGTGCAGGACACAG   41                       AAGCCGGGCGAGCATGAG   42                       AATCTGCTTGACCCCAAA   43                       GAAACCCCTGTAGCAATC   44                       GTATCAGAAACCCCTGTA   45                       GCTCGCAGATGGTATCAG   46                       GCAGGGCTCGCAGATGGT   47                       TGGGCAGGGCTCGCAGAT   48                       GACTGGGCAGGGCTCGCA   49                       CATTGGAGAAGAAGCCGA   50                       GATGACACATTGGAGAAG   51                       GCAGATGACACATTGGAG   52                       TCGAAAGCAGATGACACA   53                       GTCCAAGGGTGACATTTT   54                       CACAGCTTGTCCAAGGGT   55                       TTGGTCTCACAGCTTGTC   56                       CAGGTCTTTGGTCTCACA   57                       CTGTTGCACAACCAGGTC   58                       GTTTGTGCCTGCCTGTTG   59                       GTCTTGTTTGTGCCTGCC   60                       CCACAGACAACATCAGTC   61                       CTGGGGACCACAGACAAC   62                       TCAGCCGATCCTGGGGAC   63                       CACCACCAGGGCTCTCAG   64                       GGGATCACCACCAGGGCT   65                       GAGGATGGCAAACAGGAT   66                       ACCAGCACCAAGAGGATG   67                       TTTTGATAAAGACCAGCA   68                       TATTGGTTGGCTTCTTGG   69                       GGGTTCCTGCTTGGGGTG   70                       GTCGGGAAAATTGATCTC   71                       GATCGTCGGGAAAATTGA   72                       GGAGCCAGGAAGATCGTC   73                       TGGAGCCAGGAAGATCGT   74                       TGGAGCAGCAGTGTTGGA   75                       GTAAAGTCTCCTGCACTG   76                       TGGCATCCATGTAAAGTC   77                       CGGTTGGCATCCATGTAA   78                       CTCTTTGCCATCCTCCTG   79                       CTGTCTCTCCTGCACTGA   80                       GGTGCAGCCTCACTGTCT   81                       AACTGCCTGTTTGCCCAC   82                       CTTCTGCCTGCACCCCTG   83                       ACTGACTGGGCATAGCTC   84                      
 
     [0304] The sequences shown in Table 6-a, above, and Table 6-b, below, are in a 5′ to 3′ direction. This is reversed with respect to 3′ to 5′ direction shown in the .seq files of Example 3. For synthesis purposes, the .seq files are generated reading from 3′ to 5′. This allows for aligning all of the 3′ most “A” nucleosides together, all of the 3′ most “G” nucleosides together, all of the 3′ most “C” nucleosides together and all of the 3′ most “T” nucleosides together. Thus when the first nucleoside of each particular oligonucleotide (attached to the solid support) is added to the wells on the plates, machine movement is reduced since an automatic pipette can move in a linear manner down one row and up another on the 96 well plate.  
     [0305] The location of the well holding each particular oligonucleotides is indicated by row and column. There are eight rows designated A to G and twelve columns designated 1 to 12 in a typical 96 well format plate. Any particular well location is indicated by its “Well No.” which is indicated by the combination of the row and the column, e.g. A08 is the well at row A, column 8.  
     [0306] In Table 6-b below, the oligonucleotides of Table 6-a are shown reordered according to the Well No. on their synthesis plate. The order shown in Table 6-b is the actually order as synthesized on an automated synthesizer taking advantage of the preferred placement of the first nucleoside according to the above alignment criteria.  
                   TABLE 6-b                          Sequences of Oligonucleotides           Targeted to CD40 Order by Synthesis Well No.                         Well No.       SEQ ID NO:                                     A01   GACCAGGCGGCAGGACCA   2           A02   AGGTGAGACCAGGCGGCA   3       A03   GCAGAGGCAGACGAACCA   5       A04   GCAAGCAGCCCCAGAGGA   6       A05   GGTCAGCAAGCAGCCCCA   7       A06   GACAGCGGTCAGCAAGCA   8       A07   GATGGACAGCGGTCAGCA   9       A08   GGTGGTTCTGGATGGACA   11       A09   GCAGTGGGTGGTTCTGGA   13       A10   CACAAAGAACAGCACTGA   14       A11   CTGGCACAAAGAACAGCA   15       A12   TCCTGGCTGGCACAAAGA   16       B01   CTGTCCTGGCTGGCACAA   17       B02   ACTCTGTGCAGTCACTCA   21       B03   TTCACCGCAAGGAAGGCA   25       B04   CTCTGTTCCAGGTGTCTA   26       B05   GTGCCAGCCTTCTTCACA   37       B06   TGCAGGACACAGCTCTCA   40       B07   AATCTGCTTGACCCCAAA   43       B08   GTATCAGAAACCCCTGTA   45       B09   GACTGGGCAGGGCTCGCA   49       B10   CATTGGAGAAGAAGCCGA   50       B11   TCGAAAGCAGATGACACA   53       B12   CAGGTCTTTGGTCTCACA   57       C01   TTTTGATAAAGACCAGCA   68       C02   GATCGTCGGGAAAATTGA   72       C03   TGGAGCAGCAGTGTTGGA   75       C04   CGGTTGGCATCCATGTAA   78       C05   CTGTCTCTCCTGCACTGA   80       C06   TCTGGATGGACAGCGGTC   10       C07   CTGGTGGCAGTGTGTCTC   27       C08   GGTGCCCTTCTGCTGGAC   31       C09   ACAGGTGCAGATGGTGTC   35       C10   GAAACCCCTGTAGCAATC   44       C11   TTGGTCTCACAGCTTGTC   56       C12   CTGTTGCACAACCAGGTC   58       D01   GTCTTGTTTGTGCCTGCC   60       D02   CCACAGACAACATCAGTC   61       D03   CTGGGGACCACAGACAAC   62       D04   TCAGCCGATCCTGGGGAC   63       D05   GTCGGGAAAATTGATCTC   71       D06   GGAGCCAGGAAGATCGTC   73       D07   TGGCATCCATGTAAAGTC   77       D08   AACTGCCTGTTTGCCCAC   82       D09   ACTGACTGGGCATAGCTC   84       D10   GTGGGTGGTTCTGGATGG   12       D11   GAAGGCATTCCGTTTCAG   24       D12   GTGTCTGTTTCTGAGGTG   33       E01   TGGTGTCTGTTTCTGAGG   34       E02   GGACACAGCTCTCACAGG   39       E03   GAGCGGTGCAGGACACAG   41       E04   AAGCCGGGCGAGCATGAG   42       E05   GCTCGCAGATGGTATCAG   46       E06   GATGACACATTGGAGAAG   51       E07   GCAGATGACACATTGGAG   52       E08   GTTTGTGCCTGCCTGTTG   59       E09   CACCACCAGGGCTCTCAG   64       E10   ACCAGCACCAAGAGGATG   67       E11   TATTGGTTGGCTTCTTGG   69       E12   GGGTTCCTGCTTGGGGTG   70       F01   GTAAAGTCTCCTGCACTG   76       F02   CTCTTTGCCATCCTCCTG   79       F03   CTTCTGCCTGCACCCCTG   83       F04   CCAGGCGGCAGGACCACT   1       F05   CAGAGGCAGACGAACCAT   4       F06   CTCACCAGTTTCTGTCCT   18       F07   TCACTCACCAGTTTCTGT   19       F08   GTGCAGTCACTCACCAGT   20       F09   CAGTGAACTCTGTGCAGT   22       F10   ATTCCGTTTCAGTGAACT   23       F11   TGGGGTCGCAGTATTTGT   28       F12   GGTTGGGGTCGCAGTATT   29       G01   CTAGGTTGGGGTCGCAGT   30       G02   CTGAGGTGCCCTTCTGCT   32       G03   TTCACAGGTGCAGATGGT   36       G04   TACAGTGCCAGCCTTCTT   38       G05   GCAGGGCTCGCAGATGGT   47       G06   TGGGCAGGGCTCGCAGAT   48       G07   GTCCAAGGGTGACATTTT   54       G08   CACAGCTTGTCCAAGGGT   55       G09   GGGATCACCACCAGGGCT   65       G10   GAGGATGGCAAACAGGAT   66       G11   TGGAGCCAGGAAGATCGT   74       G12   GGTGCAGCCTCACTGTCT   81                  
 
     Example 12  
     [0307] Automated Assay of CD40 Oligonucleotide Activity  
     [0308] A. Poly(A)+ mRNA Isolation.  
     [0309] Poly(A)+ mRNA was isolated according to Miura et al. ( Clin. Chem.,  1996, 42, 1758). Briefly, for cells grown on 96-well plates, growth medium was removed from the cells and each well was washed with 200 μl cold PBS. 60 μi lysis buffer (10 mM Tris-HCl, pH 7.6, 1 mM EDTA, 0.5 M NaCl, 0.5% NP-40, 20 mM vanadyl-ribonucleoside complex) was added to each well, the plate was gently agitated and then incubated at room temperature for five minutes. 55 μl of lysate was transferred to Oligo d(T) coated 96 well plates (AGCT Inc., Irvine, Calif.). Plates were incubated for 60 minutes at room temperature, washed 3 times with 200 ml of wash buffer (10 mM Tris-HCl pH 7.6, 1 mM EDTA, 0.3 M NaCl). After the final wash, the plate was blotted on paper towels to remove excess wash buffer and then air-dried for 5 minutes. 60 ml of elution buffer (5 mM Tris-HCl pH 7.6), preheated to 70° C. was added to each well, the plate was incubated on a 90° C. plate for 5 minutes, and the eluate then transferred to a fresh 96-well plate. Cells grown on 100 mm or other standard plates may be treated similarly, using appropriate volumes of all solutions.  
     [0310] B. Total RNA Isolation  
     [0311] Total mRNA was isolated using an RNEASY 96™ kit and buffers purchased from Qiagen Inc. (Valencia Calif.) following the manufacturer&#39;s recommended procedures. Briefly, for cells grown on 96-well plates, growth medium was removed from the cells and each well was washed with 200 mL cold PBS. 100 mL Buffer RLT was added to each well and the plate vigorously agitated for 20 seconds. 100 mL of 70% ethanol was then added to each well and the contents mixed by pipetting three times up and down. The samples were then transferred to the RNEASY 96™ well plate attached to a QIAVAC™ manifold fitted with a waste collection tray and attached to a vacuum source. Vacuum was applied for 15 seconds. 1 mL of Buffer RW1 was added to each well of the RNEASY 96™ plate and the vacuum again applied for 15 seconds. 1 mL of Buffer RPE was then added to each well of the RNEASY 96™ plate and the vacuum applied for a period of 15 seconds. The Buffer RPE wash was then repeated and the vacuum was applied for an additional 10 minutes. The plate was then removed from the QIAVAC™ manifold and blotted dry on paper towels. The plate was then re-attached to the QIAVAC™ manifold fitted with a collection tube rack containing 1.2 mL collection tubes. RNA was then eluted by pipetting 60 mL water into each well, incubating 1 minute, and then applying the vacuum for 30 seconds. The elution step was repeated with an additional 60 mL water.  
     [0312] C. RT-PCR Analysis of CD40 mRNA Levels  
     [0313] Quantitation of CD40 mRNA levels was accomplished by real-time quantitative PCR using the ABI PRISM™ 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer&#39;s instructions. This is a closed-tube, non-gel-based, fluorescence detection system which allows high-throughput quantitation of polymerase chain reaction (PCR) products in real-time. As opposed to standard PCR in which amplification products are quantitated after the PCR is completed, products in real-time quantitative PCR are quantitated as they accumulate. This is accomplished by including in the PCR reaction an oligonucleotide probe that anneals specifically between the forward and reverse PCR primers, and contains two fluorescent dyes. A reporter dye (e.g., FAM or JOE, obtained from either PE-Applied Biosystems, Foster City, Calif., Operon Technologies Inc., Alameda, Calif. or Integrated DNA Technologies Inc., Coralville, Iowa) is attached to the 5′ end of the probe and a quencher dye (e.g., TAMRA, obtained from either PE-Applied Biosystems, Foster City, Calif., Operon Technologies Inc., Alameda, Calif. or Integrated DNA Technologies Inc., Coralville, Iowa) is attached to the 3′ end of the probe. When the probe and dyes are intact, reporter dye emission is quenched by the proximity of the 3′ quencher dye. During amplification, annealing of the probe to the target sequence creates a substrate that can be cleaved by the 5′-exonuclease activity of Taq polymerase. During the extension phase of the PCR amplification cycle, cleavage of the probe by Taq polymerase releases the reporter dye from the remainder of the probe (and hence from the quencher moiety) and a sequence-specific fluorescent signal is generated. With each cycle, additional reporter dye molecules are cleaved from their respective probes, and the fluorescence intensity is monitored at regular intervals by laser optics built into the ABI PRISM™ Sequence Detection System. In each assay, a series of parallel reactions containing serial dilutions of mRNA from untreated control samples generates a standard curve that is used to quantitate the percent inhibition after antisense oligonucleotide treatment of test samples.  
     [0314] Prior to quantitative PCR analysis, primer-probe sets specific to the target gene being measured are evaluated for their ability to be “multiplexed” with a GAPDH amplification reaction. In multiplexing, both the target gene and the internal standard gene GAPDH are amplified concurrently in a single sample. In this analysis, mRNA isolated from untreated cells is serially diluted. Each dilution is amplified in the presence of primer-probe sets specific for GAPDH only, target gene only (“single-plexing”), or both (multiplexing). Following PCR amplification, standard curves of GAPDH and target mRNA signal as a function of dilution are generated from both the single-plexed and multiplexed samples. If both the slope and correlation coefficient of the GAPDH and target signals generated from the multiplexed samples fall within 10% of their corresponding values generated from the single-plexed samples, the primer-probe set specific for that target is deemed multiplexable. Other methods of PCR are also known in the art.  
     [0315] PCR reagents were obtained from Invitrogen Corporation, (Carlsbad, Calif.). RT-PCR reactions were carried out by adding 20 μL PCR cocktail (2.5× PCR buffer minus MgCl 2 ; 6.6 mM MgCl 2 , 375 μM each of dATP, dCTP, dCTP and dGTP, 375 nM each of forward primer and reverse primer, 125 nM of probe, 4 Units RNAse inhibitor, 1.25 Units PLATINUM® Taq, 5 Units MuLV reverse transcriptase, and 2.5× ROX dye) to 96-well plates containing 30 μL total RNA solution (20-200 ng). The RT reaction was carried out by incubation for 30 minutes at 48° C. Following a 10 minute incubation at 95° C. to activate the PLATINUM® Taq, 40 cycles of a two-step PCR protocol were carried out: 95° C. for 15 seconds (denaturation) followed by 60° C. for 1.5 minutes (annealing/extension).  
     [0316] Gene target quantities obtained by real time RT-PCR are normalized using either the expression level of GAPDH, a gene whose expression is constant, or by quantifying total RNA using RiboGreen™ (Molecular Probes, Inc. Eugene, Oreg.). GAPDH expression is quantified by real time RT-PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RiboGreen™ RNA quantification reagent (Molecular Probes, Inc. Eugene, Oreg.). Methods of RNA quantification by RiboGreen™ are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374).  
     [0317] In this assay, 170 μL of RiboGreen™ working reagent (RiboGreen™ reagent diluted 1:350 in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5) is pipetted into a 96-well plate containing 30 μL purified, cellular RNA. The plate is read in a CytoFluor 4000 (PE Applied Biosystems) with excitation at 485 nm and emission at 530 nm.  
     [0318] For human GAPDH the PCR primers were:  
     [0319] forward primer: GAAGGTGAAGGTCGGAGTC(SEQ ID NO: 89)  
     [0320] reverse primer: GAAGATGGTGATGGGATTTC (SEQ ID NO: 90) and the PCR probe  
     [0321] was: 5′ JOE-CAAGCTTCCCGTTCTCAGCC-TAMRA 3′ (SEQ ID NO: 91) where JOE is the fluorescent reporter dye and TAMRA is the quencher dye.  
     Example 13  
     [0322] Inhibition of CD40 Expression by Phosphorothioate Oligodeoxynucleotides  
     [0323] In accordance with the present invention, a series of oligonucleotides complementary to mRNA were designed to target different regions of the human CD40 mRNA, using published sequences (GenBank accession number X60592, incorporated herein by reference as SEQ ID NO: 85). The oligonucleotides are shown in Table 7. Target sites are indicated by the beginning nucleotide numbers, as given in the sequence source reference (X60592), to which the oligonucleotide binds. All compounds in Table 7 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. Data are averages from three experiments.  
                   TABLE 7                          Inhibition of CD40 mRNA Levels           by Phosphorothioate Oligodeoxynucleotides                                         TARGET       %   SEQ ID           ISIS#   SITE   SEQUENCE   INHIB.   NO.                                             18623   18   CCAGGCGGCAGGACCAC   30.71   1           18624   20   GACCAGGCGGCAGGAC   28.09   2       18625   26   AGGTGAGACCAGGCGG   21.89   3       18626   48   CAGAGGCAGACGAACC   0.00   4       18627   49   GCAGAGGCAGACGAAC   0.00   5       18628   73   GCAAGCAGCCCCAGAG   0.00   6       18629   78   GGTCAGCAAGCAGCCCC   29.96   7       18630   84   GACAGCGGTCAGCAAGC   0.00   8       18631   88   GATGGACAGCGGTCAGC   0.00   9       18632   92   TCTGGATGGACAGCGGT   0.00   10       18633   98   GGTGGTTCTGGATGGAC   0.00   11       18634   101   GTGGGTGGTTCTGGATG   0.00   12       18635   104   GCAGTGGGTGGTTCTGG   0.00   13       18636   152   CACAAAGAACAGCACTG   0.00   14       18637   156   CTGGCACAAAGAACAGC   0.00   15       18638   162   TCCTGGCTGGCACAAAG   0.00   16       18639   165   CTGTCCTGGCTGGCACA   4.99   17       18640   176   CTCACCAGTTTCTGTCCT   0.00   18       18641   179   TCACTCACCAGTTTCTG   0.00   19       18642   185   GTGCAGTCACTCACCAG   0.00   20       18643   190   ACTCTGTGCAGTCACTC   0.00   21       18644   196   CAGTGAACTCTGTGCAG   5.30   22       18645   205   ATTCCGTTTCAGTGAAC   0.00   23       18646   211   GAAGGCATTCCGTTTCA   9.00   24       18647   222   TTCACCGCAAGGAAGGC   0.00   25       18648   250   CTCTGTTCCAGGTGTCT   0.00   26       18649   267   CTGGTGGCAGTGTGTCT   0.00   27       18650   286   TGGGGTCGCAGTATTTG   0.00   28       18651   289   GGTTGGGGTCGCAGTAT   0.00   29       18652   292   CTAGGTTGGGGTCGCAG   0.00   30       18653   318   GGTGCCCTTCTGCTGGA   19.67   31       18654   322   CTGAGGTGCCCTTCTGC   15.63   32       18655   332   GTGTCTGTTTCTGAGGT   0.00   33       18656   334   TGGTGTCTGTTTCTGAG   0.00   34       18657   345   ACAGGTGCAGATGGTGT   0.00   35       18658   348   TTCACAGGTGCAGATGG   0.00   36       18659   360   GTGCCAGCCTTCTTCAC   5.67   37       18660   364   TACAGTGCCAGCCTTCT   7.80   38       18661   391   GGACACAGCTCTCACAG   0.00   39       18662   395   TGCAGGACACAGCTCTC   0.00   40       18663   401   GAGCGGTGCAGGACAC   0.00   41       18664   416   AAGCCGGGCGAGCATG   0.00   42       18665   432   AATCTGCTTGACCCCAA   5.59   43       18666   446   GAAACCCCTGTAGCAAT   0.10   44       18667   452   GTATCAGAAACCCCTGT   0.00   45       18668   463   GCTCGCAGATGGTATCA   0.00   46       18669   468   GCAGGGCTCGCAGATGG   34.05   47       18670   471   TGGGCAGGGCTCGCAGA   0.00   48       18671   474   GACTGGGCAGGGCTCGC   2.71   49       18672   490   CATTGGAGAAGAAGCCG   0.00   50       18673   497   GATGACACATTGGAGAA   0.00   51       18674   500   GCAGATGACACATTGGA   0.00   52       18675   506   TCGAAAGCAGATGACAC   0.00   53       18676   524   GTCCAAGGGTGACATTT   8.01   54       18677   532   CACAGCTTGTCCAAGGG   0.00   55       18678   539   TTGGTCTCACAGCTTGT   0.00   56       18679   546   CAGGTCTTTGGTCTCAC   6.98   57       18680   558   CTGTTGCACAACCAGGT   18.76   58       18681   570   GTTTGTGCCTGCCTGTT   2.43   59       18682   575   GTCTTGTTTGTGCCTGCC   0.00   60       18683   590   CCACAGACAACATCAGT   0.00   61       18684   597   CTGGGGACCACAGACAA   0.00   62       18685   607   TCAGCCGATCCTGGGGA   0.00   63       18686   621   CACCACCAGGGCTCTCA   23.31   64       18687   626   GGGATCACCACCAGGGC   0.00   65       18688   657   GAGGATGGCAAACAGG   0.00   66       18689   668   ACCAGCACCAAGAGGAT   0.00   67       18690   679   TTTTGATAAAGACCAGC   0.00   68       18691   703   TATTGGTTGGCTTCTTG   0.00   69       18692   729   GGGTTCCTGCTTGGGGT   0.00   70       18693   750   GTCGGGAAAATTGATCT   0.00   71       18694   754   GATCGTCGGGAAAATTG   0.00   72       18695   765   GGAGCCAGGAAGATCGT   0.00   73       18696   766   TGGAGCCAGGAAGATCG   0.00   74       18697   780   TGGAGCAGCAGTGTTGG   0.00   75       18698   796   GTAAAGTCTCCTGCACT   0.00   76       18699   806   TGGCATCCATGTAAAGT   0.00   77       18700   810   CGGTTGGCATCCATGTA   0.00   78       18701   834   CTCTTTGCCATCCTCCTG   4.38   79       18702   861   CTGTCTCTCCTGCACTG   0.00   80       18703   873   GGTGCAGCCTCACTGTC   0.00   81       18704   910   AACTGCCTGTTTGCCCA   33.89   82       18705   954   CTTCTGCCTGCACCCCT   0.00   83       18706   976   ACTGACTGGGCATAGCT   0.00   84                  
 
     [0324] As shown in Table 7, SEQ ID NOS: 1, 2, 7, 47 and 82 demonstrated at least 25% inhibition of CD40 expression and are therefore preferred compounds of the invention.  
     Example 14  
     [0325] Inhibition of CD40 Expression by Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0326] In accordance with the present invention, a second series of oligonucleotides complementary to mRNA were designed to target different regions of the human CD40 mRNA, using published sequence X60592. The oligonucleotides are shown in Table 8. Target sites are indicated by the beginning or initial nucleotide numbers, as given in the sequence source reference (X60592), to which the oligonucleotide binds.  
     [0327] All compounds in Table 8 are chimeric oligonucleotides (“gapmers”) 18 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-O-(2-methoxyethyl) (2′-MOE) nucleotides. The intersugar (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines. Data are averaged from three experiments.  
                   TABLE 8                          Inhibition of CD40 mRNA Levels           by Chimeric Phosphorothioate Oligonucleotides                                         TARGET       %               ISIS#   SITE   SEQUENCE   Inhibition   SEQ ID                                             19211   18   CCAGGCGGCAGGACCA   75.71   1           19212   20   GACCAGGCGGCAGGAC   77.23   2       19213   26   AGGTGAGACCAGGCGG   80.82   3       19214   48   CAGAGGCAGACGAACC   23.68   4       19215   49   GCAGAGGCAGACGAAC   45.97   5       19216   73   GCAAGCAGCCCCAGAG   65.80   6       19217   78   GGTCAGCAAGCAGCCC   74.73   7       19218   84   GACAGCGGTCAGCAAG   67.21   8       19219   88   GATGGACAGCGGTCAG   65.14   9       19220   92   TCTGGATGGACAGCGGT   78.71   10       19221   98   GGTGGTTCTGGATGGAC   81.33   11       19222   101   GTGGGTGGTTCTGGATG   57.79   12       19223   104   GCAGTGGGTGGTTCTGG   73.70   13       19224   152   CACAAAGAACAGCACT   40.25   14       19225   156   CTGGCACAAAGAACAG   60.11   15       19226   162   TCCTGGCTGGCACAAAG   10.18   16       19227   165   CTGTCCTGGCTGGCACA   24.37   17       19228   176   CTCACCAGTTTCTGTCC   22.30   18       19229   179   TCACTCACCAGTTTCTG   40.64   19       19230   185   GTGCAGTCACTCACCAG   82.04   20       19231   190   ACTCTGTGCAGTCACTC   37.59   21       19232   196   CAGTGAACTCTGTGCAG   40.26   22       19233   205   ATTCCGTTTCAGTGAAC   56.03   23       19234   211   GAAGGCATTCCGTTTCA   32.21   24       19235   222   TTCACCGCAAGGAAGG   61.03   25       19236   250   CTCTGTTCCAGGTGTcT   62.19   26       19237   267   CTGGTGGCAGTGTGTCT   70.32   27       19238   286   TGGGGTCGCAGTATTTG   0.00   28       19239   289   GGTTGGGGTCGCAGTAT   19.40   29       19240   292   CTAGGTTGGGGTCGCAG   36.32   30       19241   318   GGTGCCCTTCTGCTGGA   78.91   31       19242   322   CTGAGGTGCCCTTCTGC   69.84   32       19243   332   GTGTCTGTTTCTGAGGT   63.32   33       19244   334   TGGTGTCTGTTTCTGAG   42.83   34       19245   345   ACAGGTGCAGATGGTGT   73.31   35       19246   348   TTCACAGGTGCAGATGG   47.72   36       19247   360   GTGCCAGCCTTCTTCAC   61.32   37       19248   364   TACAGTGCCAGCCTTCT   46.82   38       19249   391   GGACACAGCTCTCACAG   0.00   39       19250   395   TGCAGGACACAGCTCTC   52.05   40       19251   401   GAGCGGTGCAGGACAC   50.15   41       19252   416   AAGCCGGGCGAGCATG   32.36   42       19253   432   AATCTGCTTGACCCCAA   0.00   43       19254   446   GAAACCCCTGTAGCAAT   0.00   44       19255   452   GTATCAGAAACCCCTGT   36.13   45       19256   463   GCTCGCAGATGGTATCA   64.65   46       19257   468   GCAGGGCTCGCAGATG   74.95   47       19258   471   TGGGCAGGGCTCGCAG   0.00   48       19259   474   GACTGGGCAGGGCTCG   82.00   49       19260   490   CATTGGAGAAGAAGCC   41.31   50       19261   497   GATGACACATTGGAGA   13.81   51       19262   500   GCAGATGACACATTGG   78.48   52       19263   506   TCGAAAGCAGATGAcA   59.28   53       19264   524   GTCCAAGGGTGACATTT   70.99   54       19265   532   CACAGCTTGTCCAAGGG   0.00   55       19266   539   TTGGTCTCACAGCTTGT   45.92   56       19267   546   CAGGTCTTTGGTCTCAc   63.95   57       19268   558   CTGTTGCACAACCAGGT   82.32   58       19269   570   GTTTGTGCCTGCCTGTT   70.10   59       19270   575   GTCTTGTTTGTGCCTGC   68.95   60       19271   590   CCACAGACAACATCAGT   11.22   61       19272   597   CTGGGGACCACAGACA   9.04   62       19273   607   TCAGCCGATCCTGGGGA   0.00   63       19274   621   CACCACCAGGGCTCTCA   23.08   64       19275   626   GGGATCACCACCAGGG   57.94   65       19276   657   GAGGATGGCAAACAGG   49.14   66       19277   668   ACCAGCACCAAGAGGA   3.48   67       19278   679   TTTTGATAAAGACCAGC   30.58   68       19279   703   TATTGGTTGGCTTCTTG   49.26   69       19280   729   GGGTTCCTGCTTGGGGT   13.95   70       19281   750   GTCGGGAAAATTGATcT   54.78   71       19282   754   GATCGTCGGGAAAATTG   0.00   72       19283   765   GGAGCCAGGAAGATCG   69.47   73       19284   766   TGGAGCCAGGAAGATC   54.48   74       19285   780   TGGAGCAGCAGTGTTGG   15.17   75       19286   796   GTAAAGTCTCCTGCACT   30.62   76       19287   806   TGGCATCCATGTAAAGT   65.03   77       19288   810   CGGTTGGCATCCATGTA   34.49   78       19289   834   CTCTTTGCCATCCTCCT   41.84   79       19290   861   CTGTCTCTCCTGCACTG   25.68   80       19291   873   GGTGCAGCCTCACTGTC   76.27   81       19292   910   AACTGCCTGTTTGCCCA   63.34   82       19293   954   CTTCTGCCTGCACCCCT   0.00   83       19294   976   ACTGACTGGGCATAGCT   11.55   84                  
 
     [0328] As shown in Table 8, SEQ ID NOS: 1, 2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 15, 20, 23, 25, 26, 27, 31, 32, 33, 35, 37, 40, 41, 46, 47, 49, 52, 53, 54, 57, 58, 59, 60, 65, 71, 73, 74, 77, 81 and 82 demonstrated at least 50% inhibition of CD40 expression and are therefore preferred compounds of the invention.  
     Example 15  
     [0329] Oligonucleotide-Sensitive Sites of the CD40 Target Nucleic Acid  
     [0330] As the data presented in the preceding two examples shows, several sequences were present in preferred compounds of two distinct oligonucleotide chemistries. Specifically, compounds having SEQ ID NOS: 1, 2, 7, 47 and 82 are preferred in both instances. These compounds map to different regions of the CD40 transcript but nevertheless define accessible sites of the target nucleic acid.  
     [0331] For example, SEQ ID NOS: 1 and 2 overlap each other and both map to the 5-untranslated region (5′-UTR) of CD40. Accordingly, this region of CD40 is particularly preferred for modulation via sequence-based technologies. Similarly, SEQ ID NOS: 7 and 47 map to the open reading frame of CD40, whereas SEQ ID NO: 82 maps to the 3′-untranslated region (3′-UTR). Thus, the ORF and 3′-UTR of CD40 may be targeted by sequence-based technologies as well.  
     [0332] The reverse complements of the active CD40 compounds are easily determined by those skilled in the art and may be assembled to yield nucleotide sequences corresponding to accessible sites on the target nucleic acid. For example, the assembled reverse complement of SEQ ID NOS: 1 and 2 is represented below as SEQ ID NO: 92:  
     [0333] 5′-AGTGGTCCTGCCGCCTGGTC-3′ SEQ ID NO: 92  
     [0334] TCACCAGGACGGCGGACC-5′ SEQ ID NO: 1  
     [0335] ACCAGGACGGCGGACCAG-5′ SEQ ID NO: 2  
     [0336] Through multiple iterations of the process of the invention, more extensive “footprints” are generated. A library of this information is compiled and may be used by those skilled in the art in a variety of sequence-based technologies to study the molecular and biological functions of CD40 and to investigate or confirm its role in various diseases and disorders.  
     Example 16  
     [0337] Site Selection Program  
     [0338] In a preferred embodiment of the invention, illustrated in FIG. 20, an application is deployed which facilitates the selection process for determining the target positions of the oligos to be synthesized, or “sites.” This program is written using a three-tiered object-oriented approach. All aspects of the software described, therefore, are tightly integrated with the relational database. For this reason, explicit database read and write steps are not shown. It should be assumed that each step described includes database access. The description below illustrates one way the program can be used. The actual interface allows users to skip from process to process at will, in any order.  
     [0339] Before running the site picking program, the target must have all relevant properties computed as described previously and indicated in process step  2204 . When the site picking program is launched at process step  2206  the user is presented with a panel showing targets which have previously been selected and had their properties calculated. The user selects one target to work with at process step  2208  and proceeds to decide if any derived properties will be needed at process step  2210 . Derived properties are calculated by performing mathematical operations on combinations of pre-calculated properties as defined by the user at process step  2212 .  
     [0340] The derived properties are made available as peers with all the pre-calculated properties. The user selects one of the properties to view plotted versus target position at process step  2214 . This graph is shown above a linear representation of the target. The horizontal or position axis of both the graph and target are linked and scalable by the user. The zoom range goes from showing the full target length to showing individual target bases as letters and individual property points. The user next selects a threshold value below or above which all sites will be eliminated from future consideration at process step  2216 . The user decides whether to eliminate more sites based on any other properties at process step  2218 . If they choose to eliminate more, they return to pick another property to display at process step  2214  and threshold at process step  2216 .  
     [0341] After eliminating sites, the user selects from the remaining list by choosing any property at process step  2220  and then choosing a manual or automatic selection technique at process step  2222 . In the automatic technique, the user decides whether they want to pick from maxima or minima and the number of maxima or minima to be selected as sites at process step  2224 . The software automatically finds and picks the points. When picking manually the user must decide if they wish to use automatic peak finding at process step  2226 . If the user selects automatic peak finding, then user must click on the graphed property with the mouse at process step  2236 . The nearest maxima or minima, depending on the modifier key held down, to the selected point will be picked as the site. Without the peak finding option, the user must pick a site at process step  2238  by clicking on its position on the linear representation of target.  
     [0342] Each time a site, or group of sites, is picked, a dynamic property is calculated for all possible sites (not yet eliminated) at process step  2230 . This property indicates the nearness of the site to a picked site allowing the user to pick sites in subsequent iterations based on target coverage. After new sites are picked, the user determines if the desired number of sites has been picked. If too few sites have been picked the user returns to pick more  2220 . If too many sites have been picked, the user may eliminate them by selecting and deleting them on the target display at process step  2234 . If the correct number of sites is picked, and the user is satisfied with the set of picked sites, the user registers these sites to the database along with their name, notebook number, and page number at process step  2238 . The database time stamps this registration event.  
     Example 17  
     [0343] Site Selection Program  
     [0344] In a preferred embodiment of the invention, illustrated in FIG. 21, an application is deployed which facilitates the assignment of specific chemical structure to the complement of the sequence of the sites previously picked and facilitates the registration and ordering of these now fully defined antisense compounds. This program is written using a three-tiered object-oriented approach. All aspects of the software described, therefore, are tightly integrated with the relational database. For this reason, explicit database read and write steps are not shown, it being understood that each step described also includes appropriate database read/write access.  
     [0345] To begin using the oligonucleotide chemistry assignment program, the user launches it at process step  2302 . The user then selects from the previously selected sets of oligonucleotides at process step  2304 , registered to the database in site picker&#39;s process step  2238 . Next, the user must decide whether to manually assign the chemistry a base at a time, or run the sites through a template at process step  2306 . If the user chooses to use a template, they must determine if a desired template is available at process step  2308 . If a template is not available with the desired chemistry modifications and the correct length, the user can define one at process step  2314 .  
     [0346] To define a template, the user must select the length of the oligonucleotide the template is to define. This oligonucleotide is then represented as a bar with selectable regions. The user sets the number of regions on the oligonucleotide, and the positions and lengths of these regions by dragging them back and forth on the bar. Each region is represented by a different color.  
     [0347] For each region, the user defines the chemistry modifications for the sugars, the linkers, and the heterocycles at each base position in the region. At least four heterocycle chemistries must be given, one for each of the four possible base types (A, G, C or T or U) in the site sequence the template will be applied to. A user interface is provided to select these chemistries which show the molecular structure of each component selected and its modification name. By pushing on a pop-up list next to each of the pictures, the user may choose from a list of structures and names, those possible to put in this place. For example, the heterocycle that represents the base type G is shown as a two dimensional structure diagram. If the user clicks on the pop-up list, a row of other possible structures and names is shown. The user drags the mouse to the desired chemistry and releases the mouse. Now the newly selected molecule is displayed as the choice for G type heterocycle modifications.  
     [0348] Once the user has created a template, or selected an existing one, the software applies the template at process step  2312  to each of the complements of the sites in the list. When the templates are applied, it is possible that chemistries will be defined which are impossible to make with the chemical precursors-presently used on the automatic synthesizer. To check this, a database is maintained of all precursors previously designed, and their availability for automated synthesis. When the templates are applied, the resulting molecules are tested at process step  2316  against this database to see if they are readily synthesized.  
     [0349] If a molecule is not readily synthesized, it is added to a list that the user inspects. At process step  2318 , the user decides whether to modify the chemistry to make it compatible with the currently recognized list of available chemistries or to ignore it. To modify a chemistry, the user must use the base at a time interface at process step  2322 . The user can also choose to go directly to this step, bypassing templates all together at process step  2306 .  
     [0350] The base at a time interface at process step  2322  is very similar to the template editor at process step  2314  except that instead of specifying chemistries for regions, they are defined one base at a time. This interface also differs in that it dynamically checks to see if the design is readily synthesized as the user makes selections. In other words, each choice made limits the choices the software makes available on the pop-up selection lists. To accommodate this function, an additional choice is made available on each pop-up of “not defined.” For example, this allows the user to inhibit linker choice from restricting the sugar choices by first setting the linker to “not defined.” The user would then pick the sugar, and then pick from the remaining linker choices available.  
     [0351] Once all of the sites on the list are assigned chemistries or dropped, they are registered at process step  2324  to a commercial chemical structure database. Registering to this database makes sure the structure is unique, assigns it a new identifier if it is unique, and allows future structure and substructure searching by creating various hash-tables. The compound definition is also stored at process step  2326  to various hash tables referred to as chemistry/position tables. These allow antisense compound searching and categorization based on oligonucleotide chemistry modification sequences and equivalent base sequences.  
     [0352] The results of the registration are displayed at process step  2328  with the new IDs if they are new compounds and with the old IDs if they have been previously registered. The user next selects which of the compounds processed they wish to order for synthesis at process step  2330  and registers an order list at process step  2332  by including scientist name, notebook number and page number. The database time-stamps this entry. The user may then choose at process step  2334 , to quit the program at process step  2338 , go back to the beginning and choose a new site list to work with process step  2304 , or start the oligonucleotide ordering interface at process step  2336 .  
     Example 18  
     [0353] Gene Walk to Optimize Oligonucleotide Sequence  
     [0354] A gene walk is executed using a CD40 antisense oligonucleotide having SEQ IS NO: 5′-CTGGCACAAAGAACAGCA-3′). In effecting this gene walk, the following parameters are used:  
                                                   Gene Walk Parameter   Entered value                          Oligonucleotide Sequence ID:   15           Name of Gene Target:   CD40           Scope of Gene Walk:   20           Sequence Shift Increment:    1                      
 
     [0355] Entering these values and effecting the gene walk centered on SEQ ID NO: 15 automatically generates the following new oligonucleotides:  
               TABLE 9                          Oligonucleotide Generated By Gene Walk                     SEQ ID   Sequence                             93   GAACAGCACTGACTGT       94   AGAACAGCACTGACTG       95   AAGAACAGCACTGACT       96   AAAGAACAGCACTGAC       97   CAAAGAACAGCACTGA       98   ACAAAGAACAGCACTG       99   CACAAAGAACAGCACT       100   GCACAAAGAACAGCAC       101   GGCACAAAGAACAGCA       102   TGGCACAAAGAACAGC       15   CTGGCACAAAGAACAG       103   GCTGGCACAAAGAACA       104   GGCTGGCACAAAGAAC       105   TGGCTGGCACAAAGAA       106   CTGGCTGGCACAAAGA       107   CCTGGCTGGCACAAAG       108   TCCTGGCTGGCACAAA       109   GTCCTGGCTGGCACAA       110   TGTCCTGGCTGGCACA       111   CTGTCCTGGCTGGCAC       112   TCTGTCCTGGCTGGCAC                  
 
     [0356] The list shown above contains 20 oligonucleotide sequences directed against the CD40 nucleic acid sequence. They are ordered by the position along the CD40 sequence at which the 5′ terminus of each oligonucleotide hybridizes. Thus, the first ten oligonucleotides are single-base frame shift sequences directed against the CD40 sequence upstream of compound SEQ ID NO: 15 and the latter ten are single-base frame shift sequences directed against the CD40 sequence downstream of compound SEQ ID NO: 15.  
     Example 19  
     [0357] Automated Assay of RhoC Oligonucleotide Activity  
     [0358] RhoC, a member of the Rho subfamily of small GTPases, is a protein that has been shown to be involved in a diverse set of signaling pathways including the ultimate regulation of the dynamic organization of the cytoskeleton.  
     [0359] Oligonucleotides targeting RhoC were designed, synthesized, analyzed and assayed according to procedures outlined in U.S. Ser. No. 09/067,638. Alternatively, oligonucleotides targeting RhoC can be designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0360] RhoC probes and primers were designed to hybridize to the human RhoC sequence, using published sequence information (GenBank accession number L25081, incorporated herein by reference as SEQ ID NO: 113).  
     [0361] For RhoC the PCR primers were:  
     [0362] forward primer TGATGTCATCCTCATGTGCTTCT (SEQ ID NO: 114)  
     [0363] reverse primer CCAGGATGATGGGCACGTT (SEQ ID NO: 115) and the PCR probe  
     [0364] was: FAM-CGACAGCCCTGACAGCCTGGAAA-TAMRA (SEQ ID NO: 116) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     Example 20  
     [0365] Antisense Inhibition of RhoC Expression-Phosphorothioate Oligodeoxynucleotides  
     [0366] In accordance with the present invention, a series of oligonucleotides were designed to target different regions of the human RhoC RNA, using published sequences (GenBank accession number L25081, incorporated herein by reference as SEQ ID NO: 113). The oligonucleotides are shown in Table 10. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. L25081), to which the oligonucleotide binds. All compounds in Table 10 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. The compounds were analyzed for effect on RhoC mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 10                          Inhibition of RhoC mRNA levels           by phosphorothioate oligodeoxynucleotides                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 25304   5′ UTR   4   gagctgagatgaagtcaa   29   117           25305   5′ UTR   44   gctgaagttcccaggctg   25   118       25306   5′ UTR   47   ccggctgaagttcccagg   42   119       25307   Coding   104   ggcaccatccccaacgat   81   120       25308   Coding   105   aggcaccatccccaacga   81   121       25309   Coding   111   tcccacaggcaccatccc   70   122       25310   Coding   117   aggtcttcccacaggcac   40   123       25311   Coding   127   atgaggaggcaggtcttc   41   124       25312   Coding   139   ttgctgaagacgatgagg   23   125       25313   Coding   178   tcaaagacagtagggacg   0   126       25314   Coding   181   ttctcaaagacagtaggg   2   127       25315   Coding   183   agttctcaaagacagtag   38   128       25316   Coding   342   tgttttccaggctgtcag   59   129       25317   Coding   433   tcgtcttgcctcaggtcc   79   130       25318   Coding   439   gtgtgctcgtcttgcctc   67   131       25319   Coding   445   ctcctggtgtgctcgtct   67   132       25320   Coding   483   cagaccgaacgggctcct   65   133       25321   Coding   488   ttcctcagaccgaacggg   57   134       25322   Coding   534   actcaaggtagccaaagg   33   135       25323   Coding   566   ctcccgcactccctcctt   91   136       25324   Coding   575   ctcaaacacctcccgcac   34   137       25325   Coding   581   ggccatctcaaacacctc   64   138       25326   Coding   614   cttgttcttgcggacctg   72   139       25327   Coding   625   cccctccgacgcttgttc   66   140       25328   3′ UTR   737   gtatggagccctcaggag   60   141       25329   3′ UTR   746   gagccttcagtatggagc   63   142       25330   3′ UTR   753   gaaaatggagccttcagt   24   143       25331   3′ UTR   759   ggaactgaaaatggagcc   2   144       25332   3′ UTR   763   ggagggaactgaaaatgg   13   145       25333   3′ UTR   766   gcaggagggaactgaaaa   27   146       25334   3′ UTR   851   agggcagggcataggcgt   31   147       25335   3′ UTR   854   ggaagggcagggcatagg   21   148       25336   3′ UTR   859   catgaggaagggcagggc   0   149       25337   3′ UTR   920   taaagtgctggtgtgtga   39   150       25338   3′ UTR   939   cctgtgagccagaagtgt   69   151       25339   3′ UTR   941   ttcctgtgagccagaagt   69   152       25340   3′ UTR   945   cactttcctgtgagccag   82   153       25341   3′ UTR   948   agacactttcctgtgagc   69   154       25342   3′ UTR   966   actctgggtccctactgc   20   155       25343   3′ UTR   992   tgcagaaacaactccagg   0   156                  
 
     Example 21  
     [0367] Antisense Inhibition of RhoC Expression-Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0368] In accordance with the present invention, a second series of oligonucleotides targeted to human RhoC were synthesized. The oligonucleotide sequences are shown in Table 11. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession No. L25081), to which the oligonucleotide binds.  
     [0369] All compounds in Table 11 are chimeric oligonucleotides (“gapmers”) 18 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines.  
     [0370] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 11                          Inhibition of RhoC mRNA levels           by chimeric phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 25344   5′ UTR   4   gagctgagatgaagtcaa   0   117           25345   5′ UTR   44   gctgaagttcccaggctg   35   118       25346   5′ UTR   47   ccggctgaagttcccagg   53   119       25347   Coding   104   ggcaccatccccaacgat   50   120       25348   Coding   105   aggcaccatccccaacga   56   121       25349   Coding   111   tcccacaggcaccatccc   4   122       25350   Coding   117   aggtcttcccacaggcac   11   123       25351   Coding   127   atgaggaggcaggtcttc   6   124       25352   Coding   139   ttgctgaagacgatgagg   15   125       25353   Coding   178   tcaaagacagtagggacg   32   126       25354   Coding   181   ttctcaaagacagtaggg   7   127       25355   Coding   183   agttctcaaagacagtag   39   128       25356   Coding   342   tgttttccaggctgtcag   59   129       25357   Coding   433   tcgtcttgcctcaggtcc   48   130       25358   Coding   439   gtgtgctcgtcttgcctc   36   131       25359   Coding   445   ctcctggtgtgctcgtct   61   132       25360   Coding   483   cagaccgaacgggctcct   50   133       25361   Coding   488   ttcctcagaccgaacggg   14   134       25362   Coding   534   actcaaggtagccaaagg   32   135       25363   Coding   566   ctcccgcactccctcctt   21   136       25364   Coding   575   ctcaaacacctcccgcac   9   137       25365   Coding   581   ggccatctcaaacacctc   66   138       25366   Coding   614   cttgttcttgcggacctg   61   139       25367   Coding   625   cccctccgacgcttgttc   0   140       25368   3′ UTR   737   gtatggagccctcaggag   28   141       25369   3′ UTR   746   gagccttcagtatggagc   32   142       25370   3′ UTR   753   gaaaatggagccttcagt   0   143       25371   3′ UTR   759   ggaactgaaaatggagcc   40   144       25372   3′ UTR   763   ggagggaactgaaaatgg   45   145       25373   3′ UTR   766   gcaggagggaactgaaaa   35   146       25374   3′ UTR   851   agggcagggcataggcgt   5   147       25375   3′ UTR   854   ggaagggcagggcatagg   0   148       25376   3′ UTR   859   catgaggaagggcagggc   0   149       25377   3′ UTR   920   taaagtgctggtgtgtga   20   150       25378   3′ UTR   939   cctgtgagccagaagtgt   67   151       25379   3′ UTR   941   ttcctgtgagccagaagt   61   152       25380   3′ UTR   945   cactttcctgtgagccag   80   153       25381   3′ UTR   948   agacactttcctgtgagc   0   154       25382   3′ UTR   966   actctgggtccctactgc   0   155       25383   3′ UTR   992   tgcagaaacaactccagg   0   156                  
 
     Example 22  
     [0371] Automated Assay of Cellular Inhibitor of Apoptosis-2 Expression Oligonucleotide Activity  
     [0372] Cellular Inhibitor of Apoptosis-2 (also known as c-IAP-2, apoptosis inhibitor 2, API-2, hIAP-1, and MIHC) is a member of the inhibitor of apoptosis (IAP) family of anti-apoptotic proteins which interfere with the transmission of intracellular death signals.  
     [0373] Oligonucleotides targeting Cellular Inhibitor of Apoptosis-2 were designed, synthesized, analyzed and assayed according to procedures outlined in U.S. Ser. No. 09/067,638. Alternatively, Oligonucleotides targeting Cellular Inhibitor of Apoptosis-2 can be designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0374] Cellular Inhibitor of Apoptosis-2 probes and primers were designed to hybridize to the human Cellular Inhibitor of Apoptosis-2 sequence, using published sequences information (GenBank accession number U37546, incorporated herein by reference as SEQ ID NO: 157).  
     [0375] For Cellular Inhibitor of Apoptosis-2-the PCR primers were:  
     [0376] forward primer: GGACTCAGGTGTTGGGAATCTG (SEQ ID NO: 158)  
     [0377] reverse primer: CAAGTACTCACACCTTGGAAACCA (SEQ ID NO: 159) and the PCR  
     [0378] probe was: FAM-AGATGATCCATGGGTTCAACATGCCAA-TAMRA (SEQ ID NO: 160) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     Example 23  
     [0379] Antisense Inhibition of Cellular Inhibitor of Apoptosis-2 Expression-Phosphorothioate Oligodeoxynucleotides  
     [0380] In accordance with the present invention, a series of oligonucleotides were designed to target different regions of the human Cellular Inhibitor of Apoptosis-2 RNA, using published sequences (GenBank accession number U37546, incorporated herein by reference as SEQ ID NO: 157). The oligonucleotides are shown in Table 12. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. U37546), to which the oligonucleotide binds. All compounds in Table 12 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. The compounds were analyzed for effect on Cellular Inhibitor of Apoptosis-2 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 12                          Inhibition of Cellular Inhibitor           of Apoptosis-2 mRNA levels       by phosphorotbioate oligodeoxynucleotides                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 23412   5′ UTR   3   actgaagacattttgaat   62   161           23413   5′ UTR   37   cttagaggtacgtaaaat   29   162       23414   5′ UTR   49   gcacttuatttcttaga   70   163       23415   5′ UTR   62   attttaattagaagcact   0   164       23416   5′ UTR   139   accatatttcactgattc   70   165       23417   5′ UTR   167   ctaactcaaaggaggaaa   0   166       23418   5′ UTR   175   cacaagacctaactcaaa   27   167       23419   5′ UTR   268   gctctgctgtcaagtgtt   57   168       23420   5′ UTR   303   tgtgtgactcatgaagct   23   169       23421   5′ UTR   335   ttcagtggcattcaatca   23   170       23422   5′ UTR   357   cttctccaggctactaga   50   171       23423   5′ UTR   363   ggtcaacttctccaggct   65   172       23424   5′ UTR   437   taaaacccttcacagaag   0   173       23425   5′ UTR   525   ttaaggaagaaatacaca   0   174       23426   5′ UTR   651   gcatggctttgcttttat   0   175       23427   Coding   768   caaacgtgttggcgcttt   35   176       23428   Coding   830   agcaggaaaagtggaata   0   177       23429   Coding   1015   ttaacggaatttagactc   0   178       23430   Coding   1064   atttgttactgaagaagg   0   179       23431   Coding   1118   agagccacggaaatatcc   9   180       23432   Coding   1168   aaatcttgatttgctctg   7   181       23433   Coding   1231   gtaagtaatctggcattt   0   182       23434   Coding   1323   agcaagccactctgtctc   50   183       23435   Coding   1436   tgaagtgtcttgaagctg   0   184       23436   Coding   1580   tttgacatcatcactgtt   0   185       23437   Coding   1716   tggcttgaacttgacgga   0   186       23438   Coding   1771   tcatctcctgggctgtct   40   187       23439   Coding   1861   gcagcattaatcacagga   0   188       23440   Coding   2007   tttctctctcctcttccc   10   189       23441   Coding   2150   aacatcatgttcttgttc   9   190       23442   Coding   2273   atataacacagcttcagc   0   191       23443   Coding   2350   aattgftcttccactggt   0   192       23444   Coding   2460   aagaaggagcacaatctt   70   193       23445   3′ UTR   2604   gaaaccaaattaggataa   12   194       23446   3′ UTR   2753   tgtagtgctacctcttu   69   195       23447   3′ UTR   2779   ctgaaatutgattgaat   14   196       23448   3′ UTR   2795   tacaatttcaataatgct   38   197       23449   3′ UTR   2920   gggtctcagtatgctgcc   21   198       23450   3′ UTR   3005   ccttcgatgtataggaca   0   199       23451   3′ UTR   3040   catgtccctaaaatgtca   0   200                  
 
     Example 24  
     [0381] Antisense Inhibition of Cellular Inhibitor of Apoptosis-2 Expression-Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0382] In accordance with the present invention, a second series of oligonucleotides targeted to human Cellular Inhibitor of Apoptosis-2 were synthesized. The oligonucleotide sequences are shown in Table 13. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. U37546), to which the oligonucleotide binds.  
     [0383] All compounds in Table 13 are chimeric oligonucleotides (“gapmers”) 18 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines.  
     [0384] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 13                          Inhibition of Cellular Inhibitor           of Apoptosis-2 mRNA levels       by chimeric phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 23452   5′ UTR   3   actgaagacattttgaat   35   161           23453   5′ UTR   37   cttagaggtacgtaaaat   26   162       23454   5′ UTR   49   gcacttttatttcttaga   76   163       23455   5′ UTR   62   attttaattagaagcact   0   164       23456   5′ UTR   139   accatatttcactgauc   0   165       23457   5′ UTR   167   ctaactcaaaggaggaaa   5   166       23458   5′ UTR   175   cacaagacctaactcaaa   0   167       23459   5′ UTR   268   gctctgctgtcaagtgtt   57   168       23460   5′ UTR   303   tgtgtgactcatgaagct   67   169       23461   5′ UTR   335   ttcagtggcattcaatca   59   170       23462   5′ UTR   357   cttctccaggctactaga   0   171       23463   5′ UTR   363   ggtcaacttctccaggct   75   172       23464   5′ UTR   437   taaaacccttcacagaag   11   173       23465   5′ UTR   525   ttaaggaagaaatacaca   0   174       23466   5′ UTR   651   gcatggctttgcttttat   46   175       23467   Coding   768   caaacgtgttggcgcttt   47   176       23468   Coding   830   agcaggaaaagtggaata   39   177       23469   Coding   1015   ttaacggaatttagactc   12   178       23470   Coding   1064   atttgttactgaagaagg   34   179       23471   Coding   1118   agagccacggaaatatcc   54   180       23472   Coding   1168   aaatcttgatttgctctg   34   181       23473   Coding   1231   gtaagtaatctggcattt   0   182       23474   Coding   1323   agcaagccactctgtctc   42   183       23475   Coding   1436   tgaagtgtcttgaagctg   0   184       23476   Coding   1580   tttgacatcatcactgtt   57   185       23477   Coding   1716   tggcttgaacttgacgga   23   186       23478   Coding   1771   tcatctcctgggctgtct   66   187       23479   Coding   1861   gcagcattaatcacagga   65   188       23480   Coding   2007   tttctctctcctcttccc   0   189       23481   Coding   2150   aacatcatgttcttgttc   13   190       23482   Coding   2273   atataacacagcttcagc   0   191       23483   Coding   2350   aattgttcttccactggt   60   192       23484   Coding   2460   aagaaggagcacaatctt   65   193       23485   3′ UTR   2604   gaaaccaaattaggataa   0   194       23486   3′ UTR   2753   tgtagtgctacctctttt   73   195       23487   3′ UTR   2779   ctgaaattttgattgaat   4   196       23488   3′ UTR   2795   tacaatttcaataatgct   0   197       23489   3′ UTR   2920   gggtctcagtatgctgcc   42   198       23490   3′ UTR   3005   ccttcgatgtataggaca   71   199       23491   3′ UTR   3040   catgtccctaaaatgtca   45   200                  
 
     Example 25  
     [0385] Automated Assay of ELK-1 Oligonucleotide Activity  
     [0386] ELK-1 (also known as p62TCF) is a member of the ternary complex factor (TCF) subfamily of Ets domain proteins and utilizes a bipartite recognition mechanism mediated by both protein-DNA and protein-protein interactions. This results in gene regulation not only by direct DNA binding but also by indirect DNA binding through recruitment by other factors (Rao et al.,  Science,  1989, 244, 66-70). The formation of ternary complexes with an array of proteins allows the differential regulation of many genes. The mechanism by which ELK-1 controls various signal transduction pathways involves regulating the activity of the Egr-1, pip92, nur77 and c-fos promoters by binding to the serum response element (SRE) in these promoters in response to extracellular stimuli such as growth factors, mitogens and oncogene products (Sharrocks et al.,  Int. J. Biochem. Cell Biol.,  1997, 29, 1371-1387). ELK-1 has also been shown to mediate other functions within the cell including apoptosis.  
     [0387] Oligonucleotides targeting ELK-1 were designed, synthesized, analyzed and assayed according to procedures outlined in U.S. Ser. No. 09/067,638. Alternatively, Oligonucleotides targeting ELK-1 can be designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0388] ELK-1 probes and primers were designed to hybridize to the human ELK-1 sequence, using published sequence information (GenBank accession number M25269, incorporated herein by reference as SEQ ID NO: 201).  
     [0389] For ELK-1 the PCR primers were:  
     [0390] forward primer: GCAAGGCAATGGCCACAT (SEQ ID NO: 202)  
     [0391] reverse primer: CTCCTCTGCATCCACCAGCTT (SEQ ID NO: 203) and the PCR probe  
     [0392] was: FAM-TCTCCTGGACTTCACGGGATGGTGGT-TAMRA (SEQ ID NO: 204) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     Example 26  
     [0393] Antisense Inhibition of ELK-1 Expression-Phosphorothioate Oligodeoxynucleotides  
     [0394] In accordance with the present invention, a series of oligonucleotides were designed to target different regions of the human ELK-1 RNA, using published sequences (GenBank accession number M25269, incorporated herein by reference as SEQ ID NO: 201). The oligonucleotides are shown in Table 14. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. M25269), to which the oligonucleotide binds. All compounds in Table 14 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. The compounds were analyzed for effect on ELK-1 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 14                          Inhibition of ELK-1 mRNA levels           by phosphorothioate oligodeoxynucleotides                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 24752   5′ UTR   11   cccctgcgtttccctaca   15   205           24753   5′ UTR   50   ggtggtggtggcggtggc   29   206       24754   5′ UTR   139   ggcgttggcaatgttggc   82   207       24755   5′ UTR   167   aagttgaggctgtgtgta   0   208       24756   5′ UTR   189   aggccacggacgggtctc   92   209       24757   5′ UTR   229   gattgattcgctacgatg   71   210       24758   5′ UTR   255   gggatgcggaggagtgcg   74   211       24759   5′ UTR   289   agtgctcacgccatccca   22   212       24760   Coding   328   aaactgccacagcgtcac   64   213       24761   Coding   381   gaagtccaggagatgatg   62   214       24762   Coding   395   caccaccatcccgtgaag   88   215       24763   Coding   455   tcttgttcttgcgtagtc   62   216       24764   Coding   512   tgttcttgtcatagtagt   52   217       24765   Coding   527   tcaccttgcggatgatgt   57   218       24766   Coding   582   gagcaccctgcgacctca   72   219       24767   Coding   600   ggcgggcagtcctcagtg   82   220       24768   Coding   787   ggtgaaggtggaatagag   58   221       24769   Coding   993   tccgatttcaggtuggg   55   222       24770   Coding   1110   ttggtggtuctggcaca   67   223       24771   Coding   1132   tggagggacttctggctc   69   224       24772   Coding   1376   gcgtaggaagcagggatg   34   225       24773   Coding   1440   gtgctccagaagtgaatg   64   226       24774   Coding   1498   actggatggaaactggaa   34   227       24775   Coding   1541   ggccatccacgctgatag   74   228       24776   3′ UTR   1701   ccaccacaatcagagcat   74   229       24777   3′ UTR   1711   gatccccaccccaccaca   16   230       24778   3′ UTR   1765   tgttttctgtggaggaga   48   231       24779   3′ UTR   1790   aaacagagaagttgtgga   11   232       24780   3′ UTR   1802   gggactgacagaaaacag   0   233       24781   3′ UTR   1860   ataaataaataaaccgcc   18   234       24782   3′ UTR   1894   gttaggtcaggctcatcc   56   235       24783   3′ UTR   1974   gttctcaagccagacctc   52   236       24784   3′ UTR   1992   aataaagaaagaaaggtc   41   237       24785   3′ UTR   2006   agggcaggctgagaaata   29   238       24786   3′ UTR   2053   cttctactcacatccaaa   54   239       24787   3′ UTR   2068   caaaacaaactaactctt   24   240       24788   3′ UTR   2080   ggaataataaaacaaaac   40   241       24789   3′ UTR   2107   ttcttcctggacccctga   93   242       24790   3′ UTR   2161   ccaagggtgtgattcttc   81   243       24791   3′ UTR   2200   tgtctgagagaaaggttg   55   244                  
 
     Example 27  
     [0395] Antisense Inhibition of ELK-1 Expression-Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0396] In accordance with the present invention, a second series of oligonucleotides targeted to human ELK-1 were synthesized. The oligonucleotide sequences are shown in Table 15. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. M25269), to which the oligonucleotide binds.  
     [0397] All compounds in Table 15 are chimeric oligonucleotides (“gapmers”) 18 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines.  
     [0398] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 15                          Inhibition of ELK-1 mRNA levels           by chimeric phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 24792   5′ UTR   11   cccctgcgtttccctaca   23   205           24793   5′ UTR   50   ggtggtggtggcggtggc   80   206       24794   5′ UTR   139   ggcgttggcaatgttggc   91   207       24795   5′ UTR   167   aagttgaggctgtgtgta   27   208       24796   5′ UTR   189   aggccacggacgggtctc   79   209       24797   5′ UTR   229   gattgattcgctacgatg   69   210       24798   5′ UTR   255   gggatgcggaggagtgcg   42   211       24799   5′ UTR   289   agtgctcacgccatccca   45   212       24800   Coding   328   aaactgccacagcgtcac   57   213       24801   Coding   381   gaagtccaggagatgatg   55   214       24802   Coding   395   caccaccatcccgtgaag   41   215       24803   Coding   455   tcttgttcttgcgtagtc   80   216       24804   Coding   512   tgttcttgtcatagtagt   65   217       24805   Coding   527   tcaccttgcggatgatgt   70   218       24806   Coding   582   gagcaccctgcgacctca   64   219       24807   Coding   600   ggcgggcagtcctcagtg   67   220       24808   Coding   787   ggtgaaggtggaatagag   45   221       24809   Coding   993   tccgatttcaggtttggg   75   222       24810   Coding   1110   ttggtggtttctggcaca   82   223       24811   Coding   1132   tggagggacttctggctc   60   224       24812   Coding   1376   gcgtaggaagcagggatg   49   225       24813   Coding   1440   gtgctccagaagtgaatg   71   226       24814   Coding   1498   actggatggaaactggaa   62   227       24815   Coding   1541   ggccatccacgctgatag   78   228       24816   3′ UTR   1701   ccaccacaatcagagcat   54   229       24817   3′ UTR   1711   gatccccaccccaccaca   44   230       24818   3′ UTR   1765   tgttttctgtggaggaga   74   231       24819   3′ UTR   1790   aaacagagaagttgtgga   64   232       24820   3′ UTR   1802   gggactgacagaaaacag   16   233       24821   3′ UTR   1860   ataaataaataaaccgcc   38   234       24822   3′ UTR   1894   gttaggtcaggctcatcc   59   235       24823   3′ UTR   1974   gttctcaagccagacctc   62   236       24824   3′ UTR   1992   aataaagaaagaaaggtc   35   237       24825   3′ UTR   2006   agggcaggctgagaaata   0   238       24826   3′ UTR   2053   cttctactcacatccaaa   46   239       24827   3′ UTR   2068   caaaacaaactaactctt   38   240       24828   3′ UTR   2080   ggaataataaaacaaaac   37   241       24829   3′ UTR   2107   ttcttcctggacccctga   71   242       24830   3′ UTR   2161   ccaagggtgtgattcttc   88   243       24831   3′ UTR   2200   tgtctgagagaaaggttg   65   244                  
 
     Example 28  
     [0399] Automated Assay of Gi alpha-11 Oligonucleotide Activity  
     [0400] G-alpha-11 is a member of the Gq subfamily of G proteins whose primary function is to activate PLC-b isoforms producing second messengers and affecting intracellular calcium stores.  
     [0401] Oligonucleotides targeting Gi alpha-11 were designed, synthesized, analyzed and assayed according to procedures outlined in U.S. Ser. No. 09/067,638. Alternatively, oligonucleotides targeting Gi alpha-11 can be designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0402] G-alpha-11 probes and primers were designed to hybridize to the human G-alpha-11 sequence, using published sequence information (GenBank accession number AF011497, incorporated herein by reference as SEQ ID NO: 245). For G-alpha-11 the PCR primers were:  
     [0403] forward primer: TGACCACCTTCGAGCATCAG (SEQ ID NO: 246)  
     [0404] reverse primer: CGGTCGTAGCATTCCTGGAT (SEQ ID NO: 247) and the PCR probe  
     [0405] was: FAM-TCAGTGCCATCAAGACCCTGTGGGAG-TAMRA (SEQ ID NO: 248) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     Example 29  
     [0406] Antisense Inhibition of G-alpha-11 Expression-Phosphorothioate Oligodeoxynucleotides  
     [0407] In accordance with the present invention, a series of oligonucleotides were designed to target different regions of the human G-alpha-11 RNA, using published sequences (GenBank accession number AF011497, incorporated herein by reference as SEQ ID NO: 245). The oligonucleotides are shown in Table 16. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. AF011497), to which the oligonucleotide binds. All compounds in Table 16 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. The compounds were analyzed for effect on G-alpha-11 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 16                          Inhibition of G-alpha-11 mRNA levels           by phosphorothioate oligodeoxynucleotides                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 20576   Coding   1   gatggactccagagtcat   0   249           20577   Coding   6   gccatgatggactccaga   75   250       20578   Coding   9   cacgccatgatggactcc   0   251       20579   Coding   25   ctcatcgctcaggcaaca   61   252       20580   Coding   31   cttcacctcatcgctcag   20   253       20581   Coding   36   gactccttcacctcatcg   15   254       20582   Coding   45   atccgcttggactccttc   17   255       20583   Coding   50   cgttgatccgcttggact   0   256       20584   Coding   61   ctcgatctcggcgttgat   0   257       20585   Coding   77   cccgccgcagctgcttct   58   258       20586   Coding   106   cttgagctcgcgccgggc   31   259       20587   Coding   116   gcagcagcagcttgagct   0   260       20588   Coding   127   gcccgtgccgagcagcag   0   261       20589   Coding   146   acgtgctcttcccgctct   28   262       20590   Coding   159   atctgcttgatgaacgtg   0   263       20591   Coding   162   cgcatctgcttgatgaac   0   264       20592   Coding   184   gtagccggcgccgtggat   1   265       20593   Coding   197   tgtcctcctccgagtagc   0   266       20594   Coding   199   cttgtcctcctccgagta   79   267       20595   Coding   207   aagccgcgcttgtcctcc   56   268       20596   Coding   222   tagacgagcttggtgaag   0   269       20597   Coding   230   tgttctggtagacgagct   0   270       20598   Coding   242   tggcggtgaagatgttct   0   271       20599   Coding   258   cggatcatggcctgcatg   1   272       20600   Coding   271   cgtctccatggcccggat   49   273       20601   Coding   285   tagaggatcttgagcgtc   0   274       20602   Coding   287   tgtagaggatcttgagcg   0   275       20603   Coding   297   tgctcgtacttgtagagg   7   276       20604   Coding   306   gccttgttctgctcgtac   25   277       20605   Coding   309   ttggccttgttctgctcg   0   278       20606   Coding   319   caggagcgcattggcctt   0   279       20607   Coding   340   ctccacgtccacctcccg   69   280       20608   Coding   349   ggtcaccttctccacgtc   27   281       20609   Coding   362   gatgctcgaaggtggtca   33   282       20610   Coding   373   actgacgtactgatgctc   36   283       20611   Coding   382   cttgatggcactgacgta   78   284       20612   Coding   388   cagggtcttgatggcact   0   285       20613   Coding   409   ctggatgcccgggtcctc   0   286       20614   Coding   411   tcctggatgcccgggtcc   30   287       20615   Coding   429   cgcctgcggtcgtagcat   0   288       20616   Coding   440   gctggtactcgcgcctgc   41   289       20617   Coding   459   tacttggcagagtcggag   34   290       20618   Coding   468   gtcaggtagtacttggca   76   291       20619   Coding   479   ggtcaacgtcggtcaggt   18   292       20620   Coding   489   gtggcgatgcggtcaacg   1   293       20621   Coding   503   gcaggtagcccaaggtgg   20   294       20622   Coding   518   cgtcctgctgggtgggca   40   295       20623   Coding   544   ggtggtgggcacgcggac   0   296       20624   Coding   555   tcgatgatgccggtggtg   0   297       20625   Coding   572   ccaggtcgaaagggtact   0   298       20626   Coding   578   tgttctccaggtcgaaag   33   299       20627   Coding   584   agatgatgttctccaggt   0   300       20628   Coding   591   atccggaagatgatgttc   0   301       20629   Coding   624   ctccgctccgaccgctgg   56   302       20630   Coding   634   gatccacttcctccgctc   59   303       20631   Coding   655   tgtcacguctcaaagca   0   304       20632   Coding   663   atgatggatgtcacgttc   0   305       20633   Coding   671   cgagaaacatgatggatg   0   306       20634   Coding   682   gctgagggcgacgagaaa   75   307       20635   Coding   709   cgactccaccaggacttg   40   308       20636   Coding   726   atccggttctcgugtcc   22   309       20637   Coding   728   ccatccggttctcgttgt   19   310       20638   Coding   744   agggctttgctctcctcc   77   311       20639   Coding   754   ggtccggaacagggcttt   26   312       20640   Coding   766   gtaggtgatgatggtccg   0   313       20641   Coding   787   ggaggagttctggaacca   64   314       20642   Coding   803   tgaggaagaggatgacgg   0   315       20643   Coding   818   gcaggtccttcttgttga   6   316       20644   Coding   831   atcttgtcctccagcagg   4   317       20645   Coding   842   gcgagtacaggatcttgt   17   318       20646   Coding   858   aagtagtccaccaggtgc   0   319       20647   Coding   910   gatgaactcccgcgccgc   52   320       20648   Coding   935   ggttcaggtccacgaaca   71   321       20649   Coding   958   gtagatgatcttgtcgct   0   322       20650   Coding   972   cacgtgaagtgtgagtag   0   323       20651   Coding   993   atgttctccgtgtcggtg   0   324       20652   Coding   1014   acggccgcgaacacgaag   6   325       20653   Coding   1027   gatggtgtccttcacggc   0   326       20654   Coding   1043   tcaggttcagctgcagga   3   327       20655   Coding   1059   accagattgtactccttc   0   328                  
 
     Example 30  
     [0408] Antisense Inhibition of G-alpha-11 Expression-Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0409] In accordance with the present invention, a second series of oligonucleotides targeted to human G-alpha-11 were synthesized. The oligonucleotide sequences are shown in Table 16. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. AF011497), to which the oligonucleotide binds.  
     [0410] All compounds in Table 17 are chimeric oligonucleotides (“gapmers”) 18 nucleosides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines.  
     [0411] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 17                          Inhibition of G-alpha-11 mRNA levels           by chimeric phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 20981   Coding   1   gatggactccagagtcat   0   249           20982   Coding   6   gccatgatggactccaga   0   250       20983   Coding   9   cacgccatgatggactcc   0   251       20984   Coding   25   ctcatcgctcaggcaaca   0   252       20985   Coding   31   cttcacctcatcgctcag   2   253       20986   Coding   36   gactccttcacctcatcg   0   254       20987   Coding   45   atccgcttggactccttc   19   255       20988   Coding   50   cgttgatccgcttggact   15   256       20989   Coding   61   ctcgatctcggcgttgat   0   257       20990   Coding   77   cccgccgcagctgcttct   41   258       20991   Coding   106   cttgagctcgcgccgggc   19   259       20992   Coding   116   gcagcagcagcttgagct   23   260       20993   Coding   127   gcccgtgccgagcagcag   38   261       20994   Coding   146   acgtgctcttcccgctct   34   262       20995   Coding   159   atctgcttgatgaacgtg   56   263       20996   Coding   162   cgcatctgcttgatgaac   31   264       20997   Coding   184   gtagccggcgccgtggat   0   265       20998   Coding   197   tgtcctcctccgagtagc   42   266       20999   Coding   199   cttgtcctcctccgagta   0   267       21000   Coding   207   aagccgcgcttgtcctcc   73   268       21001   Coding   222   tagacgagcttggtgaag   0   269       21002   Coding   230   tgttctggtagacgagct   61   270       21003   Coding   242   tggcggtgaagatguct   14   271       21004   Coding   258   cggatcatggcctgcatg   84   272       21005   Coding   271   cgtctccatggcccggat   70   273       21006   Coding   285   tagaggatcttgagcgtc   39   274       21007   Coding   287   tgtagaggatcttgagcg   28   275       21008   Coding   297   tgctcgtacttgtagagg   70   276       21009   Coding   306   gccttgttctgctcgtac   76   277       21010   Coding   309   ttggccttgttctgctcg   0   278       21011   Coding   319   caggagcgcattggcctt   87   279       21012   Coding   340   ctccacgtccacctcccg   0   280       21013   Coding   349   ggtcaccttctccacgtc   69   281       21014   Coding   362   gatgctcgaaggtggtca   0   282       21015   Coding   373   actgacgtactgatgctc   69   283       21016   Coding   382   cttgatggcactgacgta   32   284       21017   Coding   388   cagggtcttgatggcact   19   285       21018   Coding   409   ctggatgcccgggtcctc   63   286       21019   Coding   411   tcctggatgcccgggtcc   56   287       21020   Coding   429   cgcctgcggtcgtagcat   73   288       21021   Coding   440   gctggtactcgcgcctgc   68   289       21022   Coding   459   tacttggcagagtcggag   50   290       21023   Coding   468   gtcaggtagtacttggca   13   291       21024   Coding   479   ggtcaacgtcggtcaggt   64   292       21025   Coding   489   gtggcgatgcggtcaacg   52   293       21026   Coding   503   gcaggtagcccaaggtgg   52   294       21027   Coding   518   cgtcctgctgggtgggca   0   295       21028   Coding   544   ggtggtgggcacgcggac   81   296       21029   Coding   555   tcgatgatgccggtggtg   48   297       21030   Coding   572   ccaggtcgaaagggtaci   61   298       21031   Coding   578   tgttctccaggtcgaaag   0   299       21032   Coding   584   agatgatgttctccaggt   0   300       21033   Coding   591   atccggaagatgatguc   0   301       21034   Coding   624   ctccgctccgaccgctgg   59   302       21035   Coding   634   gatccacttcctccgctc   17   303       21036   Coding   655   tgtcacgttctcaaagca   9   304       21037   Coding   663   atgatggatgtcacgttc   41   305       21038   Coding   671   cgagaaacatgatggatg   0   306       21039   Coding   682   gctgagggcgacgagaaa   11   307       21040   Coding   709   cgactccaccaggacttg   0   308       21041   Coding   726   atccggttctcgttgtcc   67   309       21042   Coding   728   ccatccgguctcgttgt   30   310       21043   Coding   744   agggctttgctctcctcc   61   311       21044   Coding   754   ggtccggaacagggcttt   72   312       21045   Coding   766   gtaggtgatgatggtccg   68   313       21046   Coding   787   ggaggagttctggaacca   54   314       21047   Coding   803   tgaggaagaggatgacgg   23   315       21048   Coding   818   gcaggtccttcttgttga   0   316       21049   Coding   831   atcttgtcctccagcagg   39   317       21050   Coding   842   gcgagtacaggatcttgt   74   318       21051   Coding   858   aagtagtccaccaggtgc   36   319       21052   Coding   910   gatgaactcccgcgccgc   67   320       21053   Coding   935   ggttcaggtccacgaaca   37   321       21054   Coding   958   gtagatgatcttgtcgct   64   322       21055   Coding   972   cacgtgaagtgtgagtag   37   323       21056   Coding   993   atgttctccgtgtcggtg   0   324       21057   Coding   1014   acggccgcgaacacgaag   0   325       21058   Coding   1027   gatggtgtccttcacggc   69   326       21059   Coding   1043   tcaggttcagctgcagga   0   327       21060   Coding   1059   accagattgtactccttc   0   328                  
 
     Example 31  
     [0412] Automated Assay of AKT-1 Oligonucleotide Activity  
     [0413] Akt-1 (also known as PKB alpha and RAC-PK alpha) is a member of the AKT/PKB family of serine/threonine kinases and has been shown to be involved in a diverse of signaling pathways.  
     [0414] Oligonucleotides targeting AKT-1 were designed, synthesized, analyzed and assayed according to procedures outlined in U.S. Ser. No. 09/067,638. Alternatively, oligonucleotides targeting AKT-1 can be designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0415] AKT-1 probes and primers were designed to hybridize to the human AKT-1 sequence, using published sequence information (GenBank accession number M63167, incorporated herein by reference as SEQ ID NO: 329). For Akt-1 the PCR primers were:  
     [0416] forward primer: CGTGACCATGAACGAGTTTGA (SEQ ID NO: 330)  
     [0417] reverse primer: CAGGATCACCTTGCCGAAA (SEQ ID NO: 331) and the PCR probe  
     [0418] was: FAM-CTGAAGCTGCTGGGCAAGGGCA-TAMRA (SEQ ID NO: 332) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     Example 32  
     [0419] Antisense Inhibition of Akt-1 Expression-Phosphorothioate Oligodeoxynucleotides  
     [0420] In accordance with the present invention, a series of oligonucleotides were designed to target different regions of the human Akt-1 RNA, using published sequences (GenBank accession number M63167, incorporated herein by reference as SEQ ID NO: 329). The oligonucleotides are shown in Table 18. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. M63167), to which the oligonucleotide binds. All compounds in Table 18 are oligodeoxynucleotides with phosphorothioate backbones (internucleoside linkages) throughout. The compounds were analyzed for effect on Akt-1 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 18                          Inhibition of Akt-1 mRNA levels           by phosphorothioate oligodeoxynucleotides                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   tion   NO.                                                 28880   5′ UTR   4   ccctgtgccctgtcccag   55   333           28881   5′ UTR   27   cctaagcccctggtgaca   15   334       28882   5′ UTR   62   ctttgacttctttgaccc   68   335       28883   5′ UTR   70   ggcagcccctttgacttc   53   336       28884   Coding   213   caaccctccttcacaata   24   337       28885   Coding   234   tactcccctcgtttgtgc   0   338       28886   Coding   281   tgccatcattcttgagga   65   339       28887   Coding   293   agccaatgaaggtgccat   67   340       28888   Coding   352   cacagagaagttgttgag   22   341       28889   Coding   496   agtctggatggcggttgt   49   342       28890   Coding   531   tcctcctcctcctgcttc   9   343       28891   Coding   570   cctgagttgtcactgggt   49   344       28892   Coding   666   ccgaaagtgcccttgccc   56   345       28893   Coding   744   gccacgatgacttccttc   60   346       28894   Coding   927   cggtcctcggagaacaca   0   347       28895   Coding   990   acgttcttctccgagtgc   30   348       28896   Coding   1116   gtgccgcaaaaggtcttc   66   349       28897   Coding   1125   tactcaggtgtgccgcaa   66   350       28898   Coding   1461   ggcttgaagggtgggctg   41   351       28899   Coding   1497   tcaaaatacctggtgtca   51   352       28900   Coding   1512   gccgtgaactcctcatca   56   353       28901   Coding   1541   ggtcaggtggtgtgatgg   0   354       28902   Coding   1573   ctcgctgtccacacactc   61   355       28903   3′ UTR   1671   gcctctccatccctccaa   76   356       28904   3′ UTR   1739   acagcgtggcttctctca   12   357       28905   3′ UTR   1814   ttttcttccctaccccgc   64   358       28906   3′ UTR   1819   gatagttttcttccctac   0   359       28907   3′ UTR   1831   taaaacccgcaggatagt   74   360       28908   3′ UTR   1856   ggagaacaaactggatga   0   361       28909   3′ UTR   1987   ctggctgacagagtgagg   59   362       28910   3′ UTR   1991   gcggctggctgacagagt   61   363       28911   3′ UTR   2031   cccagagagatgacagat   46   364       28912   3′ UTR   2127   gctgctgtgtgcctgcca   38   365       28913   3′ UTR   2264   cataatacacaataacaa   39   366       28914   3′ UTR   2274   atttgaacaacataatac   11   367       28915   3′ UTR   2397   aagtgctaccgtggagag   57   368       28916   3′ UTR   2407   cgaaaaggtcaagtgcta   41   369       28917   3′ UTR   2453   cagggagtcagggagggc   13   370       28918   3′ UTR   2545   aaagttgaatgttgtaaa   10   371       28919   3′ UTR   2553   aaaatactaaagttgaat   25   372                  
 
     Example 33  
     [0421] Antisense Inhibition of Akt-1 Expression-Phosphorothioate 2′-MOE Gapmer Oligonucleotides  
     [0422] In accordance with the present invention, a second series of oligonucleotides targeted to human Akt-1 were synthesized. The oligonucleotide sequences are shown in Table 19. Target sites are indicated by nucleotide numbers, as given in the sequence source reference (Genbank accession no. M63167), to which the oligonucleotide binds.  
     [0423] All compounds in Table 19 are chimeric oligonucleotides (“gapmers”) 18 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by four-nucleotide “wings.” The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the 2′-MOE wings are 5-methylcytidines.  
     [0424] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments. If present, “N.D.” indicates “no data”.  
                   TABLE 19                          Inhibition of Akt-1 mRNA levels by chimeric           phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                         %                               Inhi-               TARGET       bi-   SEQ ID       ISIS#   REGION   SITE   SEQUENCE   -tion   NO.                                                 28920   5′ UTR   4   ccctgtgccctgtcccag   88   333           28921   5′ UTR   27   cctaagcccctggtgaca   44   334       28922   5′ UTR   62   ctttgacttctttgaccc   61   335       28923   5′ UTR   70   ggcagcccctttgacttc   79   336       28924   Coding   213   caaccctccttcacaata   72   337       28925   Coding   234   tactcccctcgtttgtgc   39   338       28926   Coding   281   tgccatcattcttgagga   73   339       28927   Coding   293   agccaatgaaggtgccat   62   340       28928   Coding   352   cacagagaagttgttgag   48   341       28929   Coding   496   agtctggatggcggttgt   43   342       28930   Coding   531   tcctcctcctcctgcttc   49   343       28931   Coding   570   cctgagttgtcactgggt   71   344       28932   Coding   666   ccgaaagtgcccttgccc   64   345       28933   Coding   744   gccacgatgacttccttc   66   346       28934   Coding   927   cggtcctcggagaacaca   77   347       28935   Coding   990   acgttcttctccgagtgc   89   348       28936   Coding   1116   gtgccgcaaaaggtcttc   61   349       28937   Coding   1125   tactcaggtgtgccgcaa   74   350       28938   Coding   1461   ggcttgaagggtgggctg   54   351       28939   Coding   1497   tcaaaatacctggtgtca   78   352       28940   Coding   1512   gccgtgaactcctcatca   88   353       28941   Coding   1541   ggtcaggtggtgtgatgg   71   354       28942   Coding   1573   ctcgctgtccacacactc   83   355       28943   3′ UTR   1671   gcctctccatccctccaa   86   356       28944   3′ UTR   1739   acagcgtggcttctctca   73   357       28945   3′ UTR   1814   ttttcttccctaccccgc   77   358       28946   3′ UTR   1819   gatagttttcttccctac   43   359       28947   3′ UTR   1831   taaaacccgcaggatagt   64   360       28948   3′ UTR   1856   ggagaacaaactggatga   70   361       28949   3′ UTR   1987   ctggctgacagagtgagg   90   362       28950   3′ UTR   1991   gcggctggctgacagagt   82   363       28951   3′ UTR   2031   cccagagagatgacagat   53   364       28952   3′ UTR   2127   gctgctgtgtgcctgcca   80   365       28953   3′ UTR   2264   cataatacacaataacaa   48   366       28954   3′ UTR   2274   atttgaacaacataatac   39   367       28955   3′ UTR   2397   aagtgctaccgtggagag   38   368       28956   3′ UTR   2407   cgaaaaggtcaagtgcta   83   369       28957   3′ UTR   2453   cagggagtcagggagggc   59   370       28958   3′ UTR   2545   aaagttgaatgttgtaaa   25   371       28959   3′ UTR   2553   aaaatactaaagttgaat   45   372                  
 
     Example 34  
     [0425] Cell Culture and Oligonucleotide Treatment  
     [0426] The effect of antisense compounds on target nucleic acid expression can be tested in any of a variety of cell types provided that the target nucleic acid is present at measurable levels. This can be routinely determined using, for example, PCR or Northern blot analysis. The following cell types are provided for illustrative purposes, but other cell types can be routinely used, provided that the target is expressed in the cell type chosen. This can be readily determined by methods routine in the art, for example Northern blot analysis, ribonuclease protection assays, or RT-PCR.  
     [0427] HMVEC d Neo Cells:  
     [0428] The human microvascular endothelial cell line from neonatal dermis, HMVEC d Neo, was obtained from Cascade Biologics Inc., (Portland, Oreg.). Cells are cultured through multiple passages in Medium 131 supplemented with Microvascular Growth Supplement (MVGS) in the absence of antibiotics and antimycotics.  
     [0429] HuVEC Cells:  
     [0430] The human umbilical vein endothelial cell line HuVEC was obtained from the American Type Culture Collection (Manassas, Va.). HuVEC cells are routinely cultured in EBM (Clonetics Corporation Walkersville, Md.) supplemented with SingleQuots supplements (Clonetics Corporation, Walkersville, Md.). Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence, are maintained for up to 15 passages. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 10000 cells/ well for use in RT-PCR analysis.  
     [0431] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.  
     [0432] HepG2 Cells:  
     [0433] The human hepatoblastoma cell line HepG2 was obtained from the American Type Culture Collection (Manassas, Va.). HepG2 cells are routinely cultured in Eagle&#39;s MEM supplemented with 10% fetal calf serum, non-essential amino acids, and 1 mM sodium pyruvate (Gibco/Life Technologies, Gaithersburg, Md.). Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence. Cells were seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis.  
     [0434] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.  
     [0435] AML12 Cells:  
     [0436] The AML12 (alpha mouse liver 12) cell line was established from hepatocytes from a mouse (CD1 strain, line MT42) transgenic for human TGF alpha. Cells are cultured in a 1:1 mixture of Dulbecco&#39;s modified Eagle&#39;s medium and Ham&#39;s F12 medium with 0.005 mg/ml insulin, 0.005 mg/ml transferrin, 5 ng/ml selenium, and 40 ng/ml dexamethasone, and 90%; 10% fetal bovine serum. For subculturing, spent medium is removed and fresh media of 0.25% trypsin, 0.03% EDTA solution is added. Fresh trypsin solution (1 to 2 ml) is added and the culture is left to sit at room temperature until the cells detach.  
     [0437] Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis.  
     [0438] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.  
     [0439] Primary Mouse Hepatocytes:  
     [0440] Primary mouse hepatocytes are prepared from CD-1 mice purchased from Charles River Labs (Wilmington, Mass.) and are routinely cultured in Hepatocyte Attachment Media (Gibco) supplemented with 10% Fetal Bovine Serum (Gibco/Life Technologies, Gaithersburg, Md.), 250 nM dexamethasone (Sigma), and 10 nM bovine insulin (Sigma). Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 10000 cells/well for use in RT-PCR analysis.  
     [0441] For Northern blotting or other analyses, cells are plated onto 100 mm or other standard tissue culture plates coated with rat tail collagen (200 ug/mL) (Becton Dickinson) and treated similarly using appropriate volumes of medium and oligonucleotide.  
     [0442] b.END Cells:  
     [0443] The mouse brain endothelial cell line b.END was obtained from Dr. Werner Risau at the Max Plank Institute (Bad Nauheim, Germany). b.END cells are routinely cultured in DMEM, high glucose (Gibco/Life Technologies, Gaithersburg, Md.) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, Md.). Cells are routinely passaged by trypsinization and dilution when they reach 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 3000 cells/well for use in RT-PCR analysis.  
     [0444] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.  
     [0445] Treatment with Antisense Compounds:  
     [0446] When cells reach 65-75% confluency, they are treated with oligonucleotide. For cells grown in 96-well plates, wells are washed once with 100 μL OPTI-MEM-1™ reduced-serum medium (Invitrogen Corporation, Carlsbad, Calif.) and then treated with 130 μL of OPTI-MEM-1™ containing 3.75 μg/mL LIPOFECTIN™ (Invitrogen Corporation, Carlsbad, Calif.) and the desired concentration of oligonucleotide. Cells are treated and data are obtained in triplicate. After 4-7 hours of treatment at 37° C., the medium is replaced with fresh medium. Cells are harvested 16-24 hours after oligonucleotide treatment. The concentration of oligonucleotide used varies from cell line to cell line. To determine the optimal oligonucleotide concentration for a particular cell line, the cells are treated with a positive control oligonucleotide at a range of concentrations. For human cells the positive control oligonucleotide is selected from either ISIS 13920 (TCCGTCATCGCTCCTCAGGG, SEQ ID NO: 373) which is targeted to human H-ras, or ISIS 18078, (GTGCGCGCGAGCCCGAAATC, SEQ ID NO: 374) which is targeted to human Jun-N-terminal kinase-2 (JNK2). Both controls are 2′-O-methoxyethyl gapmers (2′-O-methoxyethyls shown in bold) with a phosphorothioate backbone. For mouse or rat cells the positive control oligonucleotide is ISIS 15770, ATGCATTCTGCCCCCAAGGA, SEQ ID NO: 375, a 2′-O-methoxyethyl gapmer (2′-O-methoxyethyls shown in bold) with a phosphorothioate backbone which is targeted to both mouse and rat c-raf. The concentration of positive control oligonucleotide that results in 80% inhibition of c-H-ras (for ISIS 13920), JNK2 (for ISIS 18078) or c-raf (for ISIS 15770) mRNA is then utilized as the screening concentration for new oligonucleotides in subsequent experiments for that cell line. If 80% inhibition is not achieved, the lowest concentration of positive control oligonucleotide that results in 60% inhibition of H-ras, JNK2 or c-raf mRNA is then utilized as the oligonucleotide screening concentration in subsequent experiments for that cell line. If 60% inhibition is not achieved, that particular cell line is deemed as unsuitable for oligonucleotide transfection experiments. The concentrations of antisense oligonucleotides used herein are from 50 nM to 300 nM.  
     Example 35  
     [0447] Design of Phenotypic Assays and in vivo Studies for Target Validation with Oligonucleotides  
     [0448] Phenotypic Assays:  
     [0449] Once target modulators have been identified by the methods disclosed herein, the compounds are further investigated in one or more phenotypic assays, each having measurable endpoints predictive of efficacy in the treatment of a particular disease state or condition.  
     [0450] Phenotypic assays, kits and reagents for their use are well known to those skilled in the art and are herein used to investigate the role and/or association of any given target in health and disease. Representative phenotypic assays, which can be purchased from any one of several commercial vendors, include those for determining cell viability, cytotoxicity, proliferation or cell survival (Molecular Probes, Eugene, Oreg.; Perkin Elmer, Boston, Mass.), protein-based assays including enzymatic assays (Panvera, LLC, Madison, Wis.; BD Biosciences, Franklin Lakes, N.J.; Oncogene Research Products, San Diego, Calif.), cell regulation, signal transduction, inflammation, oxidative processes and apoptosis (Assay Designs Inc., Ann Arbor, Mich.), triglyceride accumulation (Sigma-Aldrich, St. Louis, Mo.), angiogenesis assays, tube formation assays, cytokine and hormone assays and metabolic assays (Chemicon International Inc., Temecula, Calif.; Amersham Biosciences, Piscataway, N.J.).  
     [0451] In one non-limiting example, cells determined to be appropriate for a particular phenotypic assay (i.e., MCF-7 cells selected for breast cancer studies; adipocytes for obesity studies) are treated with target modulators identified from the in vitro studies as well as control compounds at optimal concentrations which are determined by the methods described above. At the end of the treatment period, treated and untreated cells are analyzed by one or more methods specific for the assay to determine phenotypic outcomes and endpoints.  
     [0452] Phenotypic endpoints include changes in cell morphology over time or treatment dose as well as changes in levels of cellular components such as proteins, lipids, nucleic acids, hormones, saccharrides or metals. Measurements of cellular status which include pH, stage of the cell cycle, intake or excretion of biological indicators by the cell, are also endpoints of interest.  
     [0453] Analysis of the geneotype of the cell (measurement of the expression of one or more of the genes of the cell) after treatment is also used as an indicator of the efficacy or potency of the target modulators. Hallmark genes, or those genes suspected to be associated with a specific disease state, condition, or phenotype, are measured in both treated and untreated cells.  
     [0454] In vivo Studies:  
     [0455] The individual subjects of the in vivo studies described herein are warm-blooded vertebrate animals, which includes humans.  
     [0456] The clinical trial is subjected to rigorous controls to ensure that individuals are not unnecessarily put at risk and that they are fully informed about their role in the study.  
     [0457] To account for the psychological effects of receiving treatments, volunteers are randomly given placebo or target modulator. Furthermore, to prevent the doctors from being biased in treatments, they are not informed as to whether the medication they are administering is a target modulator or a placebo. Using this randomization approach, each volunteer has the same chance of being given either the new treatment or the placebo.  
     [0458] Volunteers receive either the target modulator or placebo for eight week period with biological parameters associated with the indicated disease state or condition being measured at the beginning (baseline measurements before any treatment), end (after the final treatment), and at regular intervals during the study period. Such measurements include the levels of nucleic acid molecules encoding the target or target protein levels in body fluids, tissues or organs compared to pre-treatment levels. Other measurements include, but are not limited to, indices of the disease state or condition being treated, body weight, blood pressure, serum titers of pharmacologic indicators of disease or toxicity as well as ADME (absorption, distribution, metabolism and excretion) measurements.  
     [0459] Information recorded for each patient includes age (years), gender, height (cm), family history of disease state or condition (yes/no), motivation rating (some/moderate/great) and number and type of previous treatment regimens for the indicated disease or condition.  
     [0460] Volunteers taking part in this study are healthy adults (age 18 to 65 years) and roughly an equal number of males and females participate in the study. Volunteers with certain characteristics are equally distributed for placebo and target modulator treatment. In general, the volunteers treated with placebo have little or no response to treatment, whereas the volunteers treated with the target modulator show positive trends in their disease state or condition index at the conclusion of the study.  
     Example 36  
     [0461] Northern Blot Analysis of Target mRNA Levels  
     [0462] Eighteen hours after antisense treatment, cell monolayers are washed twice with cold PBS and lysed in 1 mL RNAZOL™ (TEL-TEST “B” Inc., Friendswood, Tex.). Total RNA is prepared following manufacturer&#39;s recommended protocols. Twenty micrograms of total RNA is fractionated by electrophoresis through 1.2% agarose gels containing 1.1% formaldehyde using a MOPS buffer system (AMRESCO, Inc. Solon, Ohio). RNA is transferred from the gel to HYBOND™-N+ nylon membranes (Amersham Pharmacia Biotech, Piscataway, N.J.) by overnight capillary transfer using a Northern/Southern Transfer buffer system (TEL-TEST “B” Inc., Friendswood, Tex.). RNA transfer is confirmed by UV visualization. Membranes are fixed by UV cross-linking using a STRATALINKER™ UV Crosslinker 2400 (Stratagene, Inc, La Jolla, Calif.) and then probed using QUICKHYB™ hybridization solution (Stratagene, La Jolla, Calif.) using manufacturer&#39;s recommendations for stringent conditions.  
     [0463] To normalize for variations in loading and transfer efficiency membranes were stripped and probed for human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) RNA (Clontech, Palo Alto, Calif.).  
     [0464] Hybridized membranes were visualized and quantitated using a PHOSPHORIMAGER™ and IMAGEQUANT™ Software V3.3 (Molecular Dynamics, Sunnyvale, Calif.). Data was normalized to GAPDH levels in untreated controls.  
     Example 37  
     [0465] Western Blot Analysis of Target Protein Levels  
     [0466] Western blot analysis (immunoblot analysis) is carried out using standard methods. Cells are harvested 16-20 h after oligonucleotide treatment, washed once with PBS, suspended in Laemmli buffer (100 ul/well), boiled for 5 minutes and loaded on a 16% SDS-PAGE gel. Gels are run for 1.5 hours at 150 V, and transferred to membrane for western blotting. Appropriate primary antibody directed to target protein is used, with a radiolabeled or fluorescently labeled secondary antibody directed against the primary antibody species. Bands are visualized using a PHOSPHORIMAGER™ (Molecular Dynamics, Sunnyvale Calif.).  
     Example 38  
     [0467] Antisense Inhibition of Human Jagged 2 Expression by Chimeric Phosphorothioate Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap  
     [0468] Jagged 2 is a member of the Notch signaling pathway which plays an essential role in cellular differentiation. It has also been implicated in hyperproliferative disorders through its influences on apoptosis and proliferation.  
     [0469] Oligonucleotides targeting Jagged 2 were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0470] Prior to quantitative PCR analysis, primer-probe sets specific to the target gene being measured are evaluated for their ability to be “multiplexed” with a GAPDH amplification reaction. In multiplexing, both the target gene and the internal standard gene GAPDH are amplified concurrently in a single sample. In this analysis, mRNA isolated from untreated cells is serially diluted. Each dilution is amplified in the presence of primer-probe sets specific for GAPDH only, target gene only (“single-plexing”), or both (multiplexing). Following PCR amplification, standard curves of GAPDH and target mRNA signal as a function of dilution are generated from both the single-plexed and multiplexed samples. If both the slope and correlation coefficient of the GAPDH and target signals generated from the multiplexed samples fall within 10% of their corresponding values generated from the single-plexed samples, the primer-probe set specific for that target is deemed multiplexable. Other methods of PCR are also known in the art.  
     [0471] PCR reagents are obtained from Invitrogen Corporation, (Carlsbad, Calif.). RT-PCR reactions are carried out by adding 20 μL PCR cocktail (2.5× PCR buffer minus MgCl 2 , 6.6 mM MgCl 2 , 375 μM each of dATP, dCTP, dCTP and dGTP, 375 nM each of forward primer and reverse primer, 125 nM of probe, 4 Units RNAse inhibitor, 1.25 Units PLATINUM® Taq, 5 Units MuLV reverse transcriptase, and 2.5× ROX dye) to 96-well plates containing 30 μL total RNA solution (20-200 ng). The RT reaction is carried out by incubation for 30 minutes at 48° C. Following a 10 minute incubation at 95° C. to activate the PLATINUM® Taq, 40 cycles of a two-step PCR protocol were carried out: 95° C. for 15 seconds (denaturation) followed by 60° C. for 1.5 minutes (annealing/extension).  
     [0472] Gene target quantities obtained by real time RT-PCR are normalized using either the expression level of GAPDH, a gene whose expression is constant, or by quantifying total RNA using RiboGreen™ (Molecular Probes, Inc. Eugene, Oreg.). GAPDH expression is quantified by real time RT-PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RiboGreen™ RNA quantification reagent (Molecular Probes, Inc. Eugene, Oreg.). Methods of RNA quantification by RiboGreen™ are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374).  
     [0473] In this assay, 170 μL of RiboGreen™ working reagent (RiboGreen™ reagent diluted 1:350 in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5) is pipetted into a 96-well plate containing 30 μL purified, cellular RNA. The plate is read in a CytoFluor 4000 (PE Applied Biosystems) with excitation at 485 nm and emission at 530 nm.  
     [0474] For human GAPDH the PCR primers were:  
     [0475] forward primer: GAAGGTGAAGGTCGGAGTC (SEQ ID NO: 89)  
     [0476] reverse primer: GAAGATGGTGATGGGATTTC (SEQ ID NO: 90) and the PCR probe  
     [0477] was: 5′ JOE-CAAGCTTCCCGTTCTCAGCC-TAMRA 3′ (SEQ ID NO: 91) where JOE is the fluorescent reporter dye and TAMRA is the quencher dye.  
     [0478] Probes and primers to human Jagged 2 were designed to hybridize to a human Jagged 2 sequence, using published sequence information (GenBank accession number NM — 002226.1, incorporated herein as SEQ ID NO: 376). For human Jagged 2 the PCR primers were: forward primer: CCCAGGGCTTCTCCGG (SEQ ID NO: 377) reverse primer: AATAGTCACCCTCCAGGTTATAGCAG (SEQ ID NO: 378) and the PCR probe was: FAM-TGGATGTCGACCTTTGTGAGCCAAGC-TAMRA (SEQ ID NO: 379) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0479] The series of oligonucleotides was designed to target different regions of the human Jagged 2 RNA, using published sequences (GenBank accession number NM — 002226.1, incorporated herein as SEQ ID NO: 376, GenBank accession number AF029778.1, incorporated herein as SEQ ID NO: 380, a genomic sequence of Jagged 2 represented by residues 104001-133000 of GenBank accession number AF111170.3, incorporated herein as SEQ ID NO: 381, and GenBank accession number BE674071.1, incorporated herein as SEQ ID NO: 382). The oligonucleotides are shown in Table 20. “Target site” indicates the first (5′-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 20 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on human Jagged 2 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments in which human neonatal dermal endothelial cells (HMVEC-d Neo cells) were cultured and treated with oligonucleotides ISIS 148702-148779 (SEQ ID NOs: 383-460 according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 20                          Inhibition Of Human Jagged 2 mRNA Levels By Chimeric Phosphorothioate           Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap                                                     TARGET                           ISIS#   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO                                                     148702   3′ UTR   376   4647   tacaaaaatgcactttcacg   79   383           148703   3′ UTR   376   4698   tggcattattcaatcaaata   0   384       148704   5′ UTR   380   2 gcgcacctgcatatgcatga   10   385       148705   Coding   380   475   gaaatagcccatgggccgcg   74   386       148706   Coding   380   487   cagctgcagctcgaaatagc   62   387       148707   Coding   380   497   gcagcgcgctcagctgcagc   63   388       148708   Coding   380   518   gcagctccccgttcacgttc   33   389       148709   Coding   380   523   gctcagcagctccccgttca   67   390       148710   Coding   380   621   tggtactccttaaggcacac   74   391       148711   Coding   380   631   caccttggcctggtactcct   72   392       148712   Coding   380   658   gccgtagctgcagggccccg   65   393       148713   Coding   380   702   ggcaggtagaaggagttgcc   49   394       148714   Coding   380   775   gacgaggcccgggtcctggt   64   395       148715   Coding   380   843   ttgtcccagtcccaggcctc   92   396       148716   Coding   380   927   aggctcttccagcggtcctc   63   397       148717   Coding   380   937   gctgaagtgcaggctcttcc   61   398       148718   Coding   380   947   ccacgtggccgctgaagtgc   54   399       148719   Coding   380   1023   ggccggcagaacttgttgca   30   400       148720   Coding   380   1068   ttgccgtactggtcgcaggt   79   401       148721   Coding   380   1078   gcaggccttgttgccgtact   63   402       148722   Coding   380   1093   catccagccgtccatgcagg   84   403       148723   Coding   380   1149   cccccgtggagcaaattaca   71   404       148724   Coding   380   1183   gtagctgcacctgcactccc   84   405       148725   Coding   380   1269   cagttgcactgccagggctc   85   406       148726   Coding   380   1279   gttggtctcacagttgcact   64   407       148727   Coding   380   1287   ccgccccagttggtctcaca   77   408       148728   Coding   380   1292   gcaggccgccccagttggtc   23   409       148729   Coding   380   1297   acagagcaggccgccccagt   72   410       148730   Coding   380   1302   ttgtcacagagcaggccgcc   81   411       148731   Coding   380   1311   ttcaggtctttgtcacagag   74   412       148732   Coding   380   1321   gccacagtagttcaggtctt   60   413       148733   Coding   380   1331   ggtggtggctgccacagtag   49   414       148734   Coding   380   1443   gaggtgcaggcgtgctcagc   63   415       148735   Coding   380   1672   cccttcacactcattggcgt   62   416       148736   Coding   380   1707   aggtttttgcaagaaaaagc   52   417       148737   Coding   380   1727   cacagtaatagccgccaatc   80   418       148738   Coding   380   1753   gatgcccttccagcccggga   75   419       148739   Coding   380   1810   gcaggtgcccccatgctgac   80   420       148740   Coding   380   1820   ccaggtccttgcaggtgccc   88   421       148741   Coding   380   1845   gggcacacacactggtaccc   71   422       148742   Coding   380   1902   gggctgctggcacacttgtc   88   423       148743   Coding   380   2100   gagcagttcttgccaccaaa   85   424       148744   Coding   380   2154   ccgcagccatcgatcactct   93   425       148745   Coding   380   2334   gtgcccccattgcggcaggg   73   426       148746   Coding   380   2474   agaagtcattgaccaggtcg   77   427       148747   Coding   380   2480   cacagtagaagtcattgacc   79   428       148748   Coding   380   2520   cgtgagtggcaggtcttgcc   68   429       148749   Coding   380   2530   ctggaactcgcgtgagtggc   56   430       148750   Coding   380   2556   ccgttgctgcaggtgtaggc   72   431       148751   Coding   380   2565   caggtgccaccgttgctgca   75   432       148752   Coding   380   2570   cgtagcaggtgccaccgttg   80   433       148753   Coding   380   2658   ttgggcaggcagctgctgtt   64   434       148754   Coding   380   2770   agggttgcagtcgttggtat   50   435       148755   Coding   380   2824   gcagcggaaccagttgacgc   75   436       148756   Coding   380   2901   ccgtaggcacagggcgagga   78   437       148757   Coding   380   2925   ttgatctcatccacacacgt   80   438       148758   Coding   380   2949   ggtgggcagctacagcgata   75   439       148759   Coding   380   3061   gcagctgttgcagtcttcca   0   440       148760   Coding   380   3071   ccaggcagcggcagctgttg   71   441       148761   Coding   380   3504   ctgctgtcaggcaggtccct   48   442       148762   Coding   380   3514   ctggatcaggctgctgtcag   61   443       148763   Coding   380   3597   tccaccttgacctcggtgac   69   444       148764   Coding   380   4059   gcgcggttgtccactttggg   59   445       148765   Stop Codon   380   4104   ccctactccttgccggcgta   80   446       148766   3′ UTR   380   4156   gacggcatggctcccaccga   75   447       148767   3′ UTR   380   4274   gaataatttatacaaggtta   62   448       148768   3′ UTR   380   4306   aatactccattgttttcagc   0   449       148769   3′ UTR   380   4359   tcatacagcgagtgccacgc   74   450       148770   3′ UTR   380   4378   caccctttgctctctccttt   67   451       148771   3′ UTR   380   4492   caccggcactttggcctgga   64   452       148772   3′ UTR   380   4538   gggtcccaccaacagccatg   83   453       148773   3′ UTR   380   4845   gaagggcacttctgaaagca   56   454       148774   3′ UTR   380   4928   acagttccgagggttctgtg   20   455       148775   Intron 5   381   15219   ctggctggatcccccacact   83   456       148776   Intron 5   381   17034   gggagcactcctggctctgc   38   457       148777   Exon: Intron   381   18740   ccatactgactgatatggca   78   458           Junction       148778 Intron: Exon   381   20082   cgacatccacctgcagggtg   70   459           Junction       148779   3′ UTR   382   242   tggcaggccccgactcaaca   69   460                  
 
     [0480] As shown in Table 20, SEQ ID NOs: 383, 386, 387, 388, 390, 391, 392, 393, 394, 395,396, 397, 398, 399, 401, 402,403, 404, 405, 406, 407, 408, 410, 411, 412, 413, 414, 415, 416, 417, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 441, 442, 443, 444, 445, 446, 447, 448, 450, 451, 452, 453, 454, 456, 458, 459 and 460 demonstrated at least 40% inhibition of human Jagged 2 expression in this assay and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 39  
     [0481] Gene Function Analysis—Caspase Assay to Determine the Effect of Modulating Jagged 2 on the Process of Apoptosis  
     [0482] With specific modulators of Jagged 2 now available, it is possible to examine the role that Jagged 2 plays in cancer.  
     [0483] Programmed cell death or apoptosis involves the activation of proteases, a family of intracellular proteases, through a cascade which leads to the cleavage of a select set of proteins. The caspase family contains at least 14 caspases, with differing substrate preferences. The caspase activity assay uses a DEVD peptide to detect activated caspases in cell culture samples. The peptide is labeled with a fluorescent molecule, 7-amino-4-trifluoromethyl coumarin (AFC). Activated caspases cleave the DEVD peptide resulting in a fluorescence shift of the AFC. Increased fluorescence is indicative of increased caspase activity. The chemotherapeutic drugs taxol, cisplatin, etoposide, gemcitabine, camptothecin, aphidicolin and 5-fluorouracil all have been shown to induce apoptosis in a caspase-dependent manner.  
     [0484] The effect of the Jagged 2 modulator ISIS 148715 (SEQ ID NO: 396) was examined in normal human mammary epithelial cells (HMECs) as well as in two breast carcinoma cell lines, MCF7 and T47D, obtained from the American Type Culture Collection (Manassas Va.). The latter two cell lines express similar genes but MCF7 cells express the tumor suppressor p53, while T47D cells are deficient in p53. MCF7 cells were routinely cultured in DMEM low glucose (Gibco/Life Technologies, Gaithersburg, Md.) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, Md.). Cells were routinely passaged by trypsinization and dilution when they reached 90% confluence. T47D cells were cultured in Gibco DMEM High glucose media supplemented with 10% FBS.  
     [0485] Cells were plated at 10,000 cells per well for HMEC cells or 20,000 cells per well for MCF-7 and T47D cells, and allowed to attach to wells overnight. Plates used were 96 well Costar plate  1603  (black sides, transparent bottom). DMEM high glucose medium, with and without phenol red, were obtained from Invitrogen (Carlsbad, Calif.). MEGM medium, with and without phenol red, were obtained from Biowhittaker (Walkersville Md.). The caspase-3 activity assay kit was obtained from Calbiochem (Cat. #HTS02).  
     [0486] Before adding to cells, the oligonucleotide cocktail was mixed thoroughly and incubated for 0.5 hrs. The Jagged 2 antisense oligonucleotide ISIS 148715 (SEQ ID NO: 396) or the mixed sequence 20-mer negative oligonucleotide control, ISIS 29848 (SEQ ID NO: 461) or the LIPOFECTIN™ only vehicle control was added (generally from a 3 μM stock of oligonucleotide) to a final concentration of 200 nM with 6 μg/ml LIPOFECTIN™. The medium was removed from the plates and the plates were tapped on sterile gauze. Each well was washed in 150 μl of PBS (150 μL HBSS for HMEC cells). The wash buffer in each well was replaced with 100 μL of the oligonucleotide/ OPTI-MEM™/LIPOFECTIN™ cocktail (this was T=0 for oligonucleotide treatment). The plates were incubated for 4 hours at 37° C., after which the medium was dumped and the plate was tapped on sterile gauze. 100 μl of full growth medium without phenol red was added to each well. After 48 hours, 50 μl of oncogene buffer (provided with Calbiochem kit) with 10 μM DTT was added to each well. 20 μl of oncogene substrate (DEVD-AFC) was added to each well. The plates were read at 400±25 nm excitation and 508±20 nm emission at t=0 and t=3 time points. The t=0×(0.8) time point was subtracted from the from the t=3 time point, and the data are shown as percent of LIPOFECTIN™-only treated cells.  
     [0487] It was thus demonstrated that the antisense modulator of Jagged 2 induces caspase activity in all three cell lines tested. The Jagged 2 oligonucleotide ISIS 148715 caused roughly a 78% reduction of Jagged 2 RNA and approximately a 5.5 fold increase in fluorescence (indicating apoptosis) when administered to HMEC cells at a 200 nM concentration. In MCF7 cells, this Jagged 2 antisense modulator reduced Jagged 2 RNA levels by approximately 50% and increased fluorescence (indicating apoptosis) by approximately 3.4 fold (200 nM concentration). Similarly, in T47D cells, Jagged 2 RNA was decreased by approximately 75% and increased fluorescence (indicating apoptosis) by 8 fold (200 nM dose of ISIS 148715). A second Jagged 2 modulator, ISIS 148744 (SEQ ID NO: 425), reduced Jagged 2 RNA to a slightly lesser extent (approx. 43% reduction) than did ISIS 148715, but also increased apoptosis by approximately 2.5 fold in MCF7 cells and 3.5 fold in T47D cells. Interestingly, ISIS 148744 did not inhibit apoptosis in the normal HMEC cells, but only in the two cancer cell lines.  
     Example 40  
     [0488] Gene Function Analysis—Cell Cycle Analysis to Determine the Effect of Modulating Jagged 2 on the Process of Apoptosis  
     [0489] Cell cycle regulation is the basis for various cancer therapies. Under some circumstances normal cells undergo growth arrest, while transformed cells undergo apoptosis and this difference can be used to protect normal cells against death caused by chemotherapeutic drugs. Disruption of cell cycle checkpoints in cancer cells can increase sensitivity to chemotherapy while cells with normal checkpoints may take refuge in G1, thus increasing the therapeutic index. ISIS 148715, an antisense modulator of Jagged 2, was tested for effects on the cell cycle in normal HMEC cells and cancer cells, both with and without p53. 72 hours after treatment with ISIS 148715, cells were stained with propidium iodide to generate a cell cycle profile using a flow cytometer. The cell cycle profile was analyzed with the ModFit program (Verity Software House, Inc., Topsham Me.). Neither LIPOFECTIN™ alone nor a panel of negative antisense controls perturbed the cell cycle. However, it was found that ISIS 148715 induced apoptosis in all three cell lines, as measured by an increase in the percentage of sub-G1 cells. In T47D cells, the percent hypodiploid cells (indicative of apoptosis) was shown to increase from approximately 4.5% for LIPOFECTIN™ control-treated cells to approximately 16% for ISIS 148715-treated cells. In MCF7 cells, the percent hypodiploid cells increased from approximately 3% (LIPOFECTIN™ only) to approximately 12.5% (ISIS 148715). In normal HMEC cells the percent diploid cells increased from approximately 2% (LIPOFECTIN™ control) to approximately-8% for cells treated with ISIS 148715. This increase in apoptosis was dose-dependent. In MCF7 cells this increase went from approximately 4% at 200 nM oligonucleotide to 8% at 300 nM oligonucleotide.  
     Example 41  
     [0490] Antisense Inhibition Of Human Transforming Growth Factor-Beta 3 Expression by Chimeric Phosphorothioate Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap  
     [0491] The transforming growth factor-beta superfamily of cytokines regulates a diverse array of physiologic functions including cell proliferation and growth, cell migration, differentiation, development, production of extracellular matrix, and the immune response. Each subgroup of this superfamily initiates a unique intracellular signaling cascade activated by ligand-induced formation and activation of specific serine/threonine kinase receptor complexes. Transforming growth factor-beta 3 is believed to have a role in healing of wounds and bone fractures, and is not expressed in healthy skin.  
     [0492] Oligonucleotides targeting human transforming growth factor-beta 3 were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0493] For human transforming growth factor-beta 3 the PCR primers were:  
     [0494] forward primer: ACCAATTACTGCTTCCGCAACT (SEQ ID NO: 463) reverse primer:  
     [0495] GATCCTGTCGGAAGTCAATGTAGA (SEQ ID NO: 464) and the PCR probe was:  
     [0496] FAM-AGGAGAACTGCTGTGTGCGCCCC-TAMRA (SEQ ID NO: 465) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0497] The series of oligonucleotides was designed to target different regions of the human transforming growth factor-beta 3 RNA, using published sequences (GenBank accession number NM — 003239.1, incorporated herein as SEQ ID NO: 462, and residues 138001-167000 of GenBank accession number AF107885, the complement of which is incorporated herein as SEQ ID NO: 466). The oligonucleotides are shown in Table 21. “Target site” indicates the first (5′-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 21 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on human transforming growth factor-beta 3 mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments in which HuVEC cells were cultured and treated with oligonucleotides 155368-155715 (SEQ ID NOs: 467-544) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 21                          Inhibition Of Human Transforming Growth Factor-Beta 3 mRNA Levels           by Chimeric Phosphorothioate Oligonucleotides       Having 2′-Moe Wings and a Deoxy Gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO.                                                     155638   5′ UTR   462   5   ttgttgtccatgtgtctaaa   69   467           155639   5′ UTR   462   76   ttcaggacttccaggaagcg   62   468       155640   5′ UTR   462   106   aggtgcatgaactcactgca   75   469       155641   5′ UTR   462   205   cggcaaggcctggagaggaa   0   470       155642   Start Codon   462   248   aagtgcatcttcatgtgtga   76   471       155643   Start Codon   462   253   tttgcaagtgcatcttcatg   87   472       155644   Coding   462   258   agccctttgcaagtgcatct   79   473       155645   Coding   462   263   accagagccctttgcaagtg   70   474       155646   Coding   462   284   aagttcagcagggccaggac   45   475       155647   Coding   462   313   aagtggacagagagaggctg   64   476       155648   Coding   462   316   tgcaagtggacagagagagg   47   477       155649   Coding   462   320   gtggtgcaagtggacagaga   85   478       155650   Coding   462   341   ttgatgtggccgaagtccaa   57   479       155651   Coding   462   346   tcttcttgatgtggccgaag   69   480       155652   Coding   462   351   cctcttcttcttgatgtggc   93   481       155653   Coding   462   356   tccaccctcttcttcttgat   70   482       155654   Coding   462   361   tggcttccaccctcttcttc   72   483       155655   Coding   462   366   cctaatggcttccaccctct   87   484       155656   Coding   462   371   tgtcccctaatggcttccac   73   485       155657   Coding   462   376   agatctgtcccctaatggct   75   486       155658   Coding   462   380   ctcaagatctgtcccctaat   72   487       155659   Coding   462   383   ttgctcaagatctgtcccct   82   488       155660   Coding   462   430   ggacgtgggtcatcaccgtt   85   489       155661   Coding   462   566   atcatgtcgaatttatggat   43   490       155662   Coding   462   572   ccctggatcatgtcgaattt   70   491       155663   Coding   462   653   tccactgaggacacattgaa   90   492       155664   Coding   462   656   ttctccactgaggacacatt   95   493       155665   Coding   462   660   atttttctccactgaggaca   90   494       155666   Coding   462   706   tgggcacccgcaagacccgg   90   495       155667   Coding   462   812   gtgggcagattcttgccacc   0   496       155668   Coding   462   860   cgcacagtgtcagtgacatc   0   497       155669   Coding   462   929   aaggtgtgacatggacagtg   93   498       155670   Coding   462   934   gctgaaaggtgtgacatgga   84   499       155671   Coding   462   939   attgggctgaaaggtgtgac   0   500       155672   Coding   462   944   tctccattgggctgaaaggt   69   501       155673   Coding   462   983   aatttgatttccatcacctc   43   502       155674   Coding   462   1022   tctccacggccatggtcatc   57   503       155675   Coding   462   1163   ttgcggaagcagtaattggt   76   504       155676   Coding   462   1269   tgagcagaagttggcatagt   69   505       155677   Coding   462   1274   gggcctgagcagaagttggc   61   506       155678   Coding   462   1279   ggcaagggcctgagcagaag   50   507       155679   Coding   462   1295   gcactgcggaggtatgggca   50   508       155680   Coding   462   1346   tcagggttcagagtgttgta   50   509       155681   Coding   462   1457   gacttcaccaccatgttgga   37   510       155682   Stop Codon   462   1478   gggtctcagctacatttaca   54   511       155683   3′ UTR   462   1562   agtgaggtttgttgcttgtg   72   512       155684   3′ UTR   462   1619   gaaacctccatctcagccat   59   513       155685   3′ UTR   462   1703   agagttcagccttcctctaa   92   514       155686   3′ UTR   462   1807   ttagggtagcccaaatccca   66   515       155687   3′ UTR   462   1834   agccattctctgcccttcct   90   516       155688   3′ UTR   462   1870   tcagatctgaagtgtcttcc   94   517       155689   3′ UTR   462   1918   tccagattccctagagcaga   72   518       155690   3′ UTR   462   1929   gtataacataatccagattc   0   519       155691   3′ UTR   462   1943   aaaatgcttgccttgtataa   79   520       155692   3′ UTR   462   1979   ctgggactttgtcttcgtaa   95   521       155693   3′ UTR   462   2030   ttgcaaaagtaatagatttg   0   522       155694   3′ UTR   462   2051   ttaattgatgtagaggacag   0   523       155695   3′ UTR   462   2082   ctggattttctccctgtagt   86   524       155696   3′ UTR   462   2093   aactgcatgacctggatttt   7   525       155697   3′ UTR   462   2112   atacagttgatgggccagga   74   526       155698   3′ UTR   462   2126   atccaaaaggcccaatacag   36   527       155699   3′ UTR   462   2151   ccaccctttcttctgcgttc   87   528       155700   3′ UTR   462   2235   gtctaaccaagtgtccaagg   92   529       155701   3′ UTR   462   2280   tgcatggaaccacaatccag   96   530       155702   3′ UTR   462   2292   atgccccaaggctgcatgga   75   531   3        155703   3′ UTR   462   2335   aatgaacacagggtcttgga   87   532       155704   3′ UTR   462   2356   cacctgcttccaggaacacc   87   533       155705   3′ UTR   462   2361   tgtagcacctgcttccagga   28   534       155706   3′ UTR   462   2407   agtcactgtgtggcacatgt   4   535       155707   3′ UTR   462   2456   agtaatattcatacttgtct   23   536       155708   3′ UTR   462   2482   atatttatttatacaaagat   0   537       155709   3′ UTR   462   2534   ctgttctagaaacaatattc   62   538       155710   Intron   466   11878   ctgctggaagcaaaggcagg   0   539       155711   Intron:   466   12956   gaggagttacctggaagagc   0   540           Exon           Junction       155712   Intron:   466   13385   gtccacctacctcttctcaa   33   541           Exon           Junction       155713   Intron   466   18442   atgccatctacatggttttt   9   542       155714   Intron:   466   21023   ttgtccacgcctgaagaagg   56   543           Exon           Junction       155715   Intron:   466   21195   ccagtctcaccggaagcagt   1   544           Exon           Junction                  
 
     [0498] As shown in Table 21, SEQ ID NOs: 467, 468, 469, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 498, 499, 501, 502, 503, 504, 505, 506, 507, 508, 509, 511, 512, 513, 514, 515, 516, 517, 518, 520, 521, 524, 526, 528, 529, 530, 531, 532, 533, 538 and 543 demonstrated at least 40% inhibition of human transforming growth factor-beta 3 expression in this assay and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 42  
     [0499] Treatment Of Angiogenic Disease: Breast Cancer  
     [0500] Breast carcinoma accounts for the most common type of tumors in women over 40 years age and is a leading cause of death. The beneficial effect on patients with breast cancer with the transforming growth factor beta 3 modulator may be shown in the following clinical trials: In a first clinical trial, 5 patients suffering from metastatic breast carcinoma are studied, who have no previous systemic treatment of metastasis (adjuvant treatment is ignored) and have facile vein access. The patients have PS 0 or 1 and may be post-menopausal. The transforming growth factor beta 3 modulator may be continuously administered parenterally, e.g. s.c. by means of a pump at the rate of e.g. 0.5 to 2 mg per 24 hours, over at least 3 days. The growth factor IGF profile is determined and the levels are found to be reduced.  
     [0501] A second clinical trial may be effected as follows: In a second trial the transforming growth factor beta 3 modulators are administered to at least 14 patients having breast cancer and the extent and duration of the response are determined.  
     [0502] Patients are included who have breast cancer as evidenced by histological biopsy (glandular analysis—EOA). They present a metastatic illness and/or loco-regional localization which is measurable and evaluable. If desired, patients are included who are resistant to other treatment to conventional therapy such as surgery, radiotherapy, other chemotherapy and/or hormone therapy.  
     [0503] The patients present at least one target (identifier), on X-ray analysis, which is measurable or evaluable such as a primitive metastatic tumor which is cutaneous or sub-cutaneous. It may be gangliar or visceral. Preferably the patients have lesions which have progressed within the month preceding the trial and have an estimated survival time of at least 3 month.  
     [0504] Preferably the trial excludes: patients in which the sole criteria for diagnosing breast cancer are biological modifications; patients administered with an embroynic carcinoma antigen pathology; patients with ascitis, a pleural effusion, a pulmonary carcinoma lymphangitis, or an osseous localization as sole metastatic manifestation; patients treated on a unique tumoral target by radiotherapy less than eight weeks before inclusion in the study (they are eligible however if evidence of progression during this time); patients with a unique cerebral localization; patients presenting another malignant tumor with the exception of a carcinoma in situ in the cervix uteri or a spino- or basocellular skin cancer; and patients not able to attend regular consultations.  
     [0505] With these exclusions the efficacy of the transforming growth factor beta 3 modulators may be followed more clearly. The transforming growth factor beta 3 modulators may be used in the method of treatment at the invention, however, in treating patients falling in the above exclusion.  
     [0506] The transforming growth factor beta 3 modulators may be administered at the same dosage as or at a lower dosage than in the first trial, but preferably in two doses, one in the morning and one in the evening. The treatment is for at least 3 months or until complete remission. The response may be followed by conventional methodology, e.g. according to IUCC response criteria, e.g. progression, stabilization, partial or complete remission. The evaluation is effected e.g. on day 0, 15, 45, 60 and 90.  
     [0507] A third clinical trial may be effected as follows: Patients with advanced breast cancer are included. The patients have progressive disease and measurable and/or evaluable parameters according to criteria of the IUCC (i.e. appearance of new lesions or growth of existing metastatic lesions) not responding to primary hormonal and/or cytotoxic therapy. As in the above indicated second clinical trial, the third trial preferably also excludes patients with previous or concurrent malignancies at other sites, with the exception of cone biopsied in situ carcinoma of the cervix uteri and adequately treated basal or squamous cell carcinoma of the skin.  
     [0508] The transforming growth factor beta 3 modulators may be administered at the same dosage as or at a lower dosage than in the second trial. Preferably the modulators are administered parenterally, e.g. subcutaneous, particularly in a continuous subcutaneous way by means of a portable syringe pump (infusion pump). Treatment is for at least 2 months or until complete remission. The response may be followed by conventional methodology e.g. according to IUCC response criteria. The evaluation is effected e.g. on day 0, 30 and 60. All lesions are measured at each assessment or when multiple lesions are present, a representative number of 5 lesions may be selected for measurement. Regression of the lesions is the sum of the products of the diameters of each individual lesion or those selected for study, which decreases by 50% or more.  
     Example 43  
     [0509] Treatment Of Angiogenic Disease: Melanoma  
     [0510] In an in vivo test, Meth-A sarcoma and melanoma cells (1.times. 10.sup.6) are inoculated subcutaneously in 0.1 ml saline in the same position of the dorsal skin of C3H mice (n=20). On the same day, the mice receive orally either transforming growth factor beta 3 modulators, at 100 mg per kg in body weight, suspended in 300 uL of olive oil (n=10) or 300 uL olive oil alone (n=10). This treatment is carried out every day and the diameter of the tumors is monitored every second day. On day 12 the mice are sacrificed and the tumor weights are measured.  
     [0511] Meth-A sarcoma tumor growth in mice treated with transforming growth factor beta 3 modulators is slower than in control mice. The weight (grams) of both the Meth-A sarcoma and melanoma tumors on day 12 is measured, and the mice treated with transforming growth factor beta 3 modulators have lower tumor mass. In a small number of control and 2-methoxyestradiol mice, the dorsal skin, together with the tumor, are excised and the angiogenesis within the subcutaneous fascia in the control and treated mice is visualized with Indian ink. Apart from their marginally lower weight, the treated mice exhibit no apparent signs of toxicity and are all alive after 12 days of daily treatment. Transforming growth factor beta 3 modulators thus has potent pharmacological properties which may be applied in the treatment angiogenic diseases, including solid tumors.  
     Example 44  
     [0512] Methods Of Inhibiting Angiogenesis  
     [0513] Angiogenesis is the growth of new blood vessels (veins &amp; arteries) by endothelial cells. This process is important in the development of a number of human diseases, and is believed to be particularly important in regulating the growth of solid tumors. Without new vessel formation it is believed that tumors will not grow beyond a few millimeters in size. In addition to their use as anti-cancer agents, modulators of angiogenesis have potential for the treatment of diabetic retinopathy, cardiovascular disease, rheumatoid arthritis and psoriasis.  
     [0514] During the process of angiogenesis, endothelial cells perform several distinct functions, including the degradation of the extracellular matrix (ECM), migration, proliferation and the formation of tube-like structures. Various genes may regulate some of these processes in primary human umbilical vein endothelial cells (HUVECs). The below experiments employed an antisense compound as a transforming growth factor beta 3 modulators. The antisense compound comprises ISIS NO. 155701 (SEQ ID NO: 530).  
     Example 45  
     [0515] Gene Function Analysis—Matrix Metalloproteinase Assay to Determine the Effect of Modulating Transforming Growth Factor-Beta 3 on the Process of Angiogenesis  
     [0516] During angiogenesis, endothelial cells need to be able to degrade the extracellular matrix so they can migrate and form new vessels. Endothelial cells secrete matrix metalloproteinases (MMPs) in order to accomplish this degradation. MMPs are a family of zinc-dependent endopeptidases that fall into eight distinct classes: five are secreted and three are membrane-type MMPs (MT-MMPs). MMPs exert these effects by cleaving a diverse group of substrates, which include not only structural components of the extracellular matrix, but also growth-factor-binding proteins, growth-factor precursors, receptor tyrosine kinases, cell-adhesion molecules and other proteinases. In this assay the activity of MMPs secreted into the media above antisense oligonucleotide-treated HUVECs is measured.  
     [0517] MMP activity in the media above HUVECs is measured using the EnzChek Gelatinase/Collagenase Assay Kit (Molecular Probes, Eugene, Oreg.). HUVECs are plated at 3000 cells/well in 96-well plates. One day later, cells are transfected with antisense oligonucleotides according to standard published procedures (Monia et al., (1 993) J Biol Chem. Jul. 5, 1993;268(19):14514-22) with 75 nM oligonucleotide in LIPOFECTIN™ (Gibco, Grand Island, N.Y.). Antisense oligonucleotides are tested in triplicate on each 96-well plate, except for positive and negative antisense controls, which are measured up to six times per plate. Twenty hours post-transfection, MMP production is stimulated by the addition of recombinant human vascular endothelial growth factor (VEGF). Fifty hours post-transfection, a p-aminophenylmercuric acetate (APMA; Sigma-Aldrich, St. Louis, Mo.) solution is added to each well of a Corning-Costar 96-well clear bottom plate (VWR International, Brisbane, Calif.). The APMA solution is used to promote cleavage of inactive MMP precursor proteins (Nagase et al., (1991) Biomed Biochim Acta, 50(4-6):749-54). Medium above the HUVECs is then transferred to the wells. After 30 minutes, the quenched, fluorogenic MMP cleavage substrate is added, and baseline fluorescence is read immediately at 485 nm exitation/530 nm emission. Following an overnight incubation at 37° C. in the dark, plates are read again to determine the amount of fluorescence, which corresponds to MMP activity. Total protein from HUVEC lysates is used to normalize the readings, and MMP activities±standard deviation are expressed relative to transfectant-only controls.  
     [0518] The modulators caused a 52% reduction of MMP activity, as compared to MMP activity in lipid-treated cells. Thus, it is shown that transforming growth factor beta 3 modulators can prevent angiogenesis.  
     Example 46  
     [0519] Gene Function Analysis—Endothelial Tube Formation Assay to Determine the Effect of Modulating Transforming Growth Factor-Beta 3 on the Process of Angiogenesis  
     [0520] Angiogenesis is stimulated by numerous factors that promote interaction of endothelial cells with each other and with extracellular matrix molecules, resulting in the formation of capillary tubes. This morphogenic process is necessary for the delivery of oxygen to nearby tissues and plays an essential role in embryonic development, wound healing, and tumor growth. Moreover, this process can be reproduced in tissue culture by the formation of tube-like structures by endothelial cells. There are several different variations of the assay that use different matrices, such as collagen I (Kanayasu, 1991), Matrigel (Yamagishi, 1997) and fibrin (Bach, 1998) as growth substrates for the cells. In this assay, HUVECs are plated on a matrix derived from the Engelbreth-Holm-Swarm mouse tumor, which is very similar to Matrigel (Kleinman, 1986; Madri, 1986). Untreated HUVECs form tube-like structures when grown on this substrate. Loss of tube formation in-vitro has been correlated with the inhibition of angiogenesis in-vivo (Carmeliet et al., (2000) Nature 407:249-257; and Zhang et al., (2002) Cancer Research 62:2034-42), which supports the use of in-vitro tube formation as an endpoint for angiogenesis.  
     [0521] The Tube Formation Assay is performed using an In-vitro Angiogenesis Assay Kit (Chemicon International, Temecula, Calif.), or growth factor reduced Mortigel (BD Biosciences, Bedford, Mass.). Cells are plated and transfected with transforming growth factor beta 3 modulators (antisense oligonucleotides) as described for the MMP activity assay, except cells are plated at 4000 cells/well. Fifty hours post-transfection, cells are transferred to 96-well plates coated with ECMatrix™ (Chemicon International) or growth factor depleted matrigel. Under these conditions, untreated HUVECs form tube-like structures. After an overnight incubation at 37° C., treated and untreated cells are inspected by light microscopy. Individual wells are assigned discrete scores from 1 to 5 depending on the extent of tube formation. A score of 1 refers to a well with no tube formation while a score of 5 is given to wells where all cells are forming an extensive tubular network.  
     [0522] As calculated from the assigned discreet scores, cells treated with transforming growth factor beta 3 modulators had a tube formation score reduction of about 60% as compared to lipid-treated cells. Thus, it is shown that transforming growth factor beta 3 modulators can inhibit angiogenesis.  
     Example 47  
     [0523] Gene Function Analysis—Measurement of RNA Expression Levels of Angiogenic Genes to Determine the Effect of Modulating Transforming Growth Factor-Beta 3 on the Process of Angiogenesis  
     [0524] Endothelial cells must regulate the expression of many genes in order to perform the functions necessary for angiogenesis. This gene regulation has been the subject of intense scrutiny, and many genes have been identified as being important for the angiogenic phenotype. The expression levels of four genes, previously identified as being highly expressed in angiogenic endothelial cells, is measured here (Integrin beta 3, endoglin/CD105, TEM5 and MMP-14/MT-MMP1).  
     [0525] Integrin beta 3 is part of a family of heterodimeric transmembrane receptors that consist of alpha and beta subunits. Each subunit recognizes a unique set of ECM ligands, thereby allowing cells to transmit angiogenic signals from the extracellular matrix. Integrin beta 3 is prominently expressed on proliferating vascular endothelial cells, and it plays roles in allowing new blood vessels to form at tumor sites as well as allowing the epithelial cells of breast tumors to spread. Blockage of Integrin alpha 3 with monoclonal antibodies or low molecular weight antagonists inhibits blood vessel formation in a variety of in-vivo models, including tumor angiogenesis and neovascularization during oxygen-induced retinopathy.  
     [0526] Endoglin is a Transforming Growth Factor receptor-associated protein highly expressed on endothelial cells, and present on some leukemia cells and minor subsets of bone marrow cells. Its expression is upregulated in endothelial cells of angiogenic tissues and is therefore used as a prognostic indicator in various tumors. Endoglin functions as an ancillary receptor influencing binding of the Transforming Growth Factor beta (TGF-beta) family of ligands to signaling receptors, thus mediating cell survival. Mutations of the endoglin gene result in a genetic disease of the vasculature-Hereditary Haemorrhagic Telangiectasia (HHT), which is characterized by bleeding from malformed blood vessels. Defective signaling by different TGF-beta ligands and receptors is thought to be involved.  
     [0527] Tumor endothelial marker 5 (TEM5) is a putative 7-pass transmembrane protein (GPCR) for which EST sequence but no other information is available. The mRNA transcript, designated KIAA1531, encodes one of many tumor endothelium markers (TEMs) that display elevated expression (greater than 10-fold) during tumor angiogenesis. TEM5 is coordinately expressed with other TEMs on tumor endothelium in humans and mice.  
     [0528] MMP-14, a membrane-type MMP (MT-MMP) covalently linked to the cell membrane, is involved in matrix detachment and migration. MMP-14 is thought to promote tumor angiogenesis; antibodies directed against the catalytic domain of MMP-14 block endothelial-cell migration, invasion and capillary tube formation in-vitro. MMP-14 can degrade the fibrin matrix that surrounds newly formed vessels potentially allowing the endothelial cells to invade further into the tumor tissue. MMP-14 null mice have impaired angiogenesis during development, further demonstrating the role of MMP-14 in angiogenesis.  
     [0529] Cells are plated and transfected as described for the MMP activity assay. Twenty hours post-transfection, cells are stimulated with recombinant human VEGF. Total RNA is harvested 52 hours post-transfection, and the amount of total RNA from each sample is determined using a Ribogreen Assay (Molecular Probes, Eugene, Oreg.). Real-time PCR is performed on the total RNA using primer/probe sets for four Angiogenic Hallmark Genes: integrin beta 3, endoglin, Tumor endothelial marker 5 (TEM5) and Matrix Metalloproteinase 14 (MMP14/MTI-MMP). Expression levels for each gene are normalized to total RNA, and values are expressed relative to controls.  
     [0530] Cells treated with transforming growth factor beta 3 modulators had the following mRNAs reduced as compared to mRNAs of the controls: Integrin beta 3 mRNA was 88% of the control, endoglin mRNA was 74% of the control, TEM5 mRNA was 91% of the control, and MMP14/MT1-MMP was 86% of the control.  
     Example 48  
     [0531] Antisense Inhibition Of Human Apolipoprotein B mRNA Expression by Chimeric Phosphorothioate Oligonucleotides having 2′-Moe Wings and a Deoxy Gap  
     [0532] Lipoproteins are globular, micelle-like particles that consist of a non-polar core of acylglycerols and cholesteryl esters surrounded by an amphiphilic coating of protein, phospholipid and cholesterol. Apolipoprotein B is a large glycoprotein that serves an indispensable role in the assembly and secretion of lipids and in the transport and receptor-mediated uptake and delivery of distinct classes of lipoproteins. Elevated plasma levels of the ApoB-100-containing lipoprotein Lp(a) are associated with increased risk for atherosclerosis and its manifestations, which may include hypercholesterolemia, myocardial infarction, and thrombosis.  
     [0533] Oligonucleotides targeting human apolipoprotein B were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0534] Probes and primers to human apolipoprotein B were designed to hybridize to a human apolipoprotein B sequence, using published sequence information (GenBank accession number NM — 000384.1, incorporated herein as SEQ ID NO: 545). For human apolipoprotein B the PCR primers were: forward primer:  
     [0535] TGCTAAAGGCACATATGGCCT (SEQ ID NO: 546) reverse primer:  
     [0536] CTCAGGTTGGACTCTCCATTGAG (SEQ ID NO: 547) and the PCR probe was: FAM-CTTGTCAGAGGGATCCTAACACTGGCCG-TAMRA (SEQ ID NO: 548) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0537] The series of oligonucleotides was designed to target different regions of the human apolipoprotein B RNA, using published sequence information (GenBank accession number NM — 000384.1, incorporated herein as SEQ ID NO: 545). The oligonucleotides are shown in Table 22. “Target site” indicates the first (5′-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 22 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on human apolipoprotein B mRNA levels in HepG2 cells by quantitative real-time PCR as described on other examples herein. Data are averages from two experiments in which HepG2 cells were cultured and treated with oligonucleotides 147780-147833 (SEQ ID NOs: 549-602) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 22                          Inhibition Of Human Apolipoprotein B mRNA Levels           By Chimeric Phosphorothioate Oligonucleotides       Having 2′-Moe Wings and a Deoxy Gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO                                                     147780   5′ UTR   545   1   ccgcaggtcccggtgggaat   40   549           147781   5′ UTR   545   21   accgagaagggcactcagcc   35   550       147782   5′ UTR   545   71   gcctcggcctcgcggccctg   67   551       147783   Start Codon   545   114   tccatcgccagctgcggtgg   N.D.   552       147784   Coding   545   151   cagcgccagcagcgccagca   70   553       147785   Coding   545   181   gcccgccagcagcagcagca   29   554       147786   Coding   545   321   cttgaatcagcagtcccagg   34   555       147787   Coding   545   451   cttcagcaaggctttgccct   N.D.   556       147788   Coding   545   716   tttctgttgccacattgccc   95   557       147789   Coding   545   911   ggaagaggtgttgctccttg   24   558       147790   Coding   545   951   tgtgctaccatcccatactt   33   559       147791   Coding   545   1041   tcaaatgcgaggcccatctt   N.D.   560       147792   Coding   545   1231   ggacacctcaatcagctgtg   26   561       147793   Coding   545   1361   tcagggccaccaggtaggtg   N.D.   562       147794   Coding   545   1561   gtaatcttcatccccagtgc   47   563       147795   Coding   545   1611   tgctccatggtttggcccat   N.D.   564       147796   Coding   545   1791   gcagccagtcgcttatctcc   8   565       147797   Coding   545   2331   gtatagccaaagtggtccac   N.D.   566       147798   Coding   545   2496   cccaggagctggaggtcatg   N.D.   567       147799   Coding   545   2573   ttgagcccttcctgatgacc   N.D.   568       147800   Coding   545   2811   atctggaccccactcctagc   N.D.   569       147801   Coding   545   2842   cagacccgactcgtggaaga   38   570       147802   Coding   545   3367   gccctcagtagattcatcat   N.D.   571       147803   Coding   545   3611   gccatgccaccctcttggaa   N.D.   572       147804   Coding   545   3791   aacccacgtgccggaaagtc   N.D.   573       147805   Coding   545   3841   actcccagatgccttctgaa   N.D.   574       147806   Coding   545   4281   atgtggtaacgagcccgaag   100   575       147807   Coding   545   4391   ggcgtagagacccatcacat   25   576       147808   Coding   545   4641   gtgttaggatccctctgaca   N.D.   577       147809   Coding   545   5241   cccagtgatagctctgtgag   60   578       147810   Coding   545   5355   atttcagcatatgagcccat   0   579       147811   Coding   545   5691   ccctgaaccttagcaacagt   N.D.   580       147812   Coding   545   5742   gctgaagccagcccagcgat   N.D.   581       147813   Coding   545   5891   acagctgcccagtatgttct   N.D.   582       147814   Coding   545   7087   cccaataagatttataacaa   34   583       147815   Coding   545   7731   tggcctaccagagacaggta   45   584       147816   Coding   545   7841   tcatacgtttagcccaatct   100   585       147817   Coding   545   7901   gcatggtcccaaggatggtc   0   586       147818   Coding   545   8491   agtgatggaagctgcgatac   30   587       147819   Coding   545   9181   atgagcatcatgcctcccag   N.D.   588       147820   Coding   545   9931   gaacacatagccgaatgccg   100   589       147821   Coding   545   10263   gtggtgccctctaatttgta   N.D.   590       147822   Coding   545   10631   cccgagaaagaaccgaaccc   N.D.   591       147823   Coding   545   10712   tgccctgcagcttcactgaa   19   592       147824   Coding   545   11170   gaaatcccataagctcttgt   N.D.   593       147825   Coding   545   12301   agaagctgcctcttcttccc   72   594       147826   Coding   545   12401   tcagggtgagccctgtgtgt   80   595       147827   Coding   545   12471   ctaatggccccttgataaac   13   596       147828   Coding   545   12621   acgttatccttgagtccctg   12   597       147829   Coding   545   12741   tatatcccaggtttccccgg   64   598       147830   Coding   545   12801   acctgggacagtaccgtccc   N.D.   599       147831   3′ UTR   545   13921   ctgcctactgcaaggctggc   0   600       147832   3′ UTR   545   13991   agagaccttccgagccctgg   N.D.   601       147833   3′ UTR   545   14101   atgatacacaataaagactc   25   602                  
 
     [0538] As shown in Table 22, SEQ ID NOs: 549, 550, 551, 553, 555, 557, 559, 563, 570, 575, 578, 583, 584, 585, 587, 589, 594, 595 and 598 demonstrated at least 30% inhibition of human apolipoprotein B expression in this assay and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention. As apolipoprotein B exists in two forms in mammals (ApoB-48 and ApoB-100) which are colinear at the amino terminus, antisense oligonucleotides targeting nucleotides 1-6530 hybridize to both forms, while those targeting nucleotides 6531-14121 are specific to the long form of apolipoprotein B.  
     Example 49  
     [0539] Antisense Inhibition of Human Apolipoprotein B mRNA Expression by Chimeric Phosphorothioate Oligonucleotides having 2′-Moe Wings and a Deoxy Gap-Dose Response Study  
     [0540] In accordance with the present invention, a subset of the antisense oligonucleotides in Example 48 were further investigated in dose-response studies. Treatment doses were 50, 150 and 250 nM. The compounds were analyzed for their effect on human apolipoprotein B mRNA levels in HepG2 cells by quantitative real-time PCR as described in other examples herein. Data are averages from two-experiments and are shown in Table 23.  
               TABLE 23                          Inhibition of Human Apolipoprotein B mRNA       Levels By Chimeric Phosphorothioate       Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap                         Percent Inhibition                                     ISIS #   50 nM   150 nM   250 nM                                                 147788   54   63   72           147806   23   45   28           147816   25   81   65           147820   10   0   73                      
 
     Example 50  
     [0541] Antisense Inhibition of Mouse Apolipoprotein B Expression by Chimeric Phosphorothioate Oligonucleotides having 2′-Moe Wings and a Deoxy Gap  
     [0542] Oligonucleotides targeting mouse apolipoprotein B were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0543] Probes and primers to mouse apolipoprotein B were designed to hybridize to a mouse apolipoprotein B sequence, using published sequence information (GenBank accession number M35186.1, incorporated herein as SEQ ID NO: 603). For mouse apolipoprotein B the PCR primers were: forward primer: CGTGGGCTCCAGCATTCTA (SEQ ID NO: 604) reverse primer: AGTCATTTCTGCCTTTGCGTC (SEQ ID NO: 605) and the PCR probe was: FAM-CCAATGGTCGGGCACTGCTCAA-TAMRA (SEQ ID NO: 606) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0544] The series of oligonucleotides was designed to target different regions of the mouse apolipoprotein B RNA, using published sequence information (GenBank accession number M35186.1, incorporated herein as SEQ ID NO: 603). The oligonucleotides are shown in Table 24. “Target site” indicates the first (5′-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 24 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on mouse apolipoprotein B mRNA levels in primary hepatocytes by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments in which AML12 cells were cultured and treated with oligonucleotides 147475-147778 (SEQ ID NOs: 607-659) according to the protocol outlined in Example 34. If present, “N.D.” “no data”.  
                   TABLE 24                          Inhibition of Mouse Apolipoprotein B mRNA Levels           by Chimeric Phosphorothioate Oligonucleotides       Having 2′-Moe Wings and a Deoxy Gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO                                                     147475   Coding   603   13   attgtatgtgagaggtgagg   79   607           147476   Coding   603   66   gaggagattggatcttaagg   13   608       147477   Coding   603   171   cttcaaattgggactctcct   N.D.   609       147478   Coding   603   211   tccaggaattgagcttgtgc   78   610       147479   Coding   603   238   ttcaggactggaggatgagg   N.D.   611       147480   Coding   603   291   tctcaccctcatgctccatt   54   612       147481   Coding   603   421   tgactgtcaagggtgagctg   24   613       147482   Coding   603   461   gtccagcctaggaacactca   59   614       147483   Coding   603   531   atgtcaatgccacatgtcca   N.D.   615       147484   Coding   603   581   ttcatccgagaagttgggac   49   616       147485   Coding   603   601   atttgggacgaatgtatgcc   64   617       147486   Coding   603   711   agttgaggaagccagattca   N.D.   618       147487   Coding   603   964   ttcccagtcagctttagtgg   73   619       147488   Coding   603   1023   agcttgcttgttgggcacgg   72   620       147489   Coding   603   1111   cctatactggcttctatgtt   5   621       147490   Coding   603   1191   tgaactccgtgtaaggcaag   N.D.   622       147491   Coding   603   1216   gagaaatccttcagtaaggg   71   623       147492   Coding   603   1323   caatggaatgcttgtcactg   68   624       147493   Coding   603   1441   gcttcattataggaggtggt   41   625       147494   Coding   603   1531   acaactgggatagtgtagcc   84   626       147495   Coding   603   1631   gttaggaccagggattgtga   0   627       147496   Coding   603   1691   accatggaaaactggcaact   19   628       147497   Coding   603   1721   tgggaggaaaaacttgaata   N.D.   629       147498   Coding   603   1861   tgggcaacgatatctgattg   0   630       147499   Coding   603   1901   ctgcagggcgtcagtgacaa   29   631       147500   Coding   603   1932   gcatcagacgtgatgttccc   N.D.   632       147501   Coding   603   2021   cttggttaaactaatggtgc   18   633       147502   Coding   603   2071   atgggagcatggaggttggc   16   634       147503   Coding   603   2141   aatggatgatgaaacagtgg   26   635       147504   Coding   603   2201   atcaatgcctcctgttgcag   N.D.   636       147505   Coding   603   2231   ggaagtgagactttctaagc   76   637       147506   Coding   603   2281   aggaaggaactcttgatatt   58   638       147507   Coding   603   2321   attggcttcattggcaacac   81   639       147759   Coding   603   1   aggtgaggaagttggaattc   19   640       147760   Coding   603   121   ttgttccctgaagttgttac   N.D.   641       147761   Coding   603   251   gttcatggattccttcagga   45   642       147762   Coding   603   281   atgctccattctcacatgct   46   643       147763   Coding   603   338   tgcgactgtgtctgatttcc   34   644       147764   Coding   603   541   gtccctgaagatgtcaatgc   97   645       147765   Coding   603   561   aggcccagttccatgaccct   59   646       147766   Coding   603   761   ggagcccacgtgctgagatt   59   647       147767   Coding   603   801   cgtccttgagcagtgcccga   5   648       147768   Coding   603   1224   cccatatggagaaatccttc   24   649       147769   Coding   603   1581   catgcctggaagccagtgtc   89   650       147770   Coding   603   1741   gtgttgaatcccttgaaatc   67   651       147771   Coding   603   1781   ggtaaagttgcccatggctg   68   652       147772   Coding   603   1841   gttataaagtccagcattgg   78   653       147773   Coding   603   1931   catcagacgtgatgttccct   85   654       147774   Coding   603   1956   tggctagtttcaatcccctt   84   655       147775   Coding   603   2002   ctgtcatgactgccctttac   52   656       147776   Coding   603   2091   gcttgaagttcattgagaat   92   657       147777   Coding   603   2291   ttcctgagaaaggaaggaac   N.D.   658       147778   Coding   603   2331   tcagatatacattggcttca   14   659                  
 
     [0545] As shown in Table 24, SEQ ID NOs: 607, 610, 612, 614, 617, 619, 620, 623, 624, 626, 637, 638, 639, 645, 646, 647, 650, 651, 652, 653, 654, 655, 656 and 657 demonstrated at least 50% inhibition of mouse apolipoprotein B expression in this assay and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 51  
     [0546] Antisense Inhibition of Mouse Apolipoprotein B mRNA Expression by Chimeric Phosphorothioate Oligonucleotides having 2′-Moe Wings and a Deoxy Gap-Dose Response Study  
     [0547] In accordance with the present invention, a subset of the antisense oligonucleotides in Example 50 were further investigated in dose-response studies. Treatment doses were 50, 150 and 300 nM. The compounds were analyzed for their effect on mouse apolipoprotein B mRNA levels in mouse primary hepatocytes by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments and are shown in Table 25.  
               TABLE 25                          Inhibition of Mouse Apolipoprotein B mRNA       Levels by Chimeric Phosphorothioate       Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap                         Percent Inhibition                                     ISIS #   50 nM   150 nM   300 nM                                                 147483   56   88   89           147764   48   84   90           147769   3   14   28           147776   0   17   44                      
 
     Example 52  
     [0548] Target Validation—Effects of Antisense Inhibition of Apolipoprotein B (ISIS 147764) in C57BL/6 Mice: Lean Animals vs. High Fat Fed Animals  
     [0549] C57BL/6 mice, a strain reported to be susceptible to hyperlipidemia-induced atherosclerotic plaque formation were used in the following studies to evaluate antisense oligonucleotides as potential lipid lowering compounds in lean versus high fat fed mice.  
     [0550] Male C57BL/6 mice were divided into two matched groups; (1) wild-type control animals (lean animals) and (2) animals receiving a high fat diet (60% kcal fat). Control animals received saline treatment and were maintained on a normal rodent diet. After overnight fasting, mice from each group were dosed intraperitoneally every three days with saline or 50 mg/kg ISIS 147764 (SEQ ID NO: 645) for six weeks. At study termination and forty eight hours after the final injections, animals were sacrificed and evaluated for target mRNA levels in liver, cholesterol and triglyceride levels, liver enzyme levels and serum glucose levels. The results of the comparative studies are shown in Table 26.  
               TABLE 26                          Effects of ISIS 147764 Treatment On Apolipoprotein       B mRNA, Cholesterol, Lipid, Triglyceride, Liver Enzyme       And Glucose Levels in Lean and High Fat Mice.                         Percent Change                                 Treatment   Lipoproteins       Liver Enzymes                                                         Group   MRNA   CHOL   VLDL   LDL   HDL   TRIG   AST   ALT   GLUC               Lean-control   −73   −63   No   −64   −44   −34   Slight   No change   No change                   change               decrease       High Fat   −87   −67   No   −87   −65   No change   Slight   Slight   −28       Group           change               decrease   increase                  
 
     [0551] It is evident from these data that treatment with ISIS 147764 lowered cholesterol as well as LDL and HDL lipoproteins and serum glucose in both lean and high fat mice and that the effects demonstrated are, in fact, due to the inhibition of apolipoprotein B expression as supported by the decrease in mRNA levels. No significant changes in liver enzyme levels were observed, indicating that the antisense oligonucleotide was not toxic to either treatment group.  
     Example 53  
     [0552] Target Validation—Effects Of Antisense Inhibition Of Apolipoprotein B (ISIS 147764) on High Fat Fed Mice; 6 Week Timecourse Study  
     [0553] A 6-week timecourse study was performed to further investigate the effects of ISIS 147764 on lipid and glucose metabolism in high fat fed mice.  
     [0554] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of treatment with the antisense oligonucleotide, ISIS 147764. Control animals received saline treatment (50 mg/kg). A subset of animals received a daily oral dose (20 mg/kg) atorvastatin calcium (Lipitor®, Pfizer Inc.). All mice, except atorvastatin-treated animals, were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks. Serum cholesterol and lipoproteins were analyzed at 0, 2 and 6 week interim timepoints. At study termination, animals were sacrificed 48 hours after the final injections and evaluated for levels of target mRNA levels in liver, cholesterol, lipoprotein, triglyceride, liver enzyme (AST and ALT) and serum glucose levels as well as body, liver, spleen and fat pad weights.  
     Example 54  
     [0555] Target Validation—Effects Of Antisense Inhibition of Apolipoprotein B (ISIS 147764) In High Fat Fed Mice-mRNA Expression in Liver  
     [0556] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on mRNA expression. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks. At study termination, animals were sacrificed 48 hours after the final injections and evaluated for levels of target mRNA levels in liver. ISIS 147764 showed a dose-response effect, reducing mRNA levels by 15, 75 and 88% at doses of 5, 25 and 50 mg/kg, respectively.  
     Example 55  
     [0557] Target Validation—Effects Of Antisense Inhibition of Apolipoprotein B (ISIS 147764) on Serum Cholesterol and Triglyceride Levels  
     [0558] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on serum cholesterol and triglyceride levels. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks.  
     [0559] Serum cholesterol levels were measured at 0, 2 and 6 weeks and this data is shown in Table 27. Values in the table are expressed as percent inhibition and are normalized to the saline control.  
     [0560] In addition to serum cholesterol, at study termination, animals were sacrificed 48 hours after the final injections and evaluated for triglyceride levels.  
     [0561] Mice treated with ISIS 147764 showed a reduction in both serum cholesterol (240 mg/dL for control animals and 225, 125 and 110 mg/dL for doses of 5, 25, and 50 mg/kg, respectively) and triglycerides (115 mg/dL for control animals and 125, 150 and 85 mg/dL for doses of 5, 25, and 50 mg/kg, respectively) to normal levels by study end.  
     [0562] These data were also compared to the effects of atorvastatin calcium at an oral dose of 20 mg/kg which showed only a minimal decrease in serum cholesterol of 20 percent at study termination.  
               TABLE 27                          Percent Inhibition of Mouse Apolipoprotein B       Cholesterol Levels by ISIS 147764                             Percent Inhibition                                     time   Saline   5 mg/kg   25 mg/kg   50 mg/kg                                         0 weeks   0   0   0   0       2 weeks   0   5   12   20       6 weeks   0   10   45   55                  
 
     Example 56  
     [0563] Target Validation—Effects Of Antisense Inhibition of Apolipoprotein B (ISIS 147764) on Lipoprotein Levels  
     [0564] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on lipoprotein (VLDL, LDL and HDL) levels. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks.  
     [0565] Lipoprotein levels were measured at 0, 2 and 6 weeks and this data is shown in Table 28. Values in the table are expressed as percent inhibition and are normalized to the saline control. Negative values indicate an observed increase in lipoprotein levels.  
     [0566] These data were also compared to the effects of atorvastatin calcium at a daily oral dose of 20 mg/kg at 0, 2 and 6 weeks.  
     [0567] These data demonstrate that at a dose of 50 mg/kg, ISIS 147764 is capable of lowering all categories of serum lipoproteins investigated to a greater extent than atorvastatin.  
               TABLE 28                          Percent Inhibition of Mouse Apolipoprotein B       Lipoprotein Levels by ISIS 147764 as       Compared to Atorvastatin                         Percent Inhibition                             Dose                                             Lipo-   Time       5   25   50   atorvastatin       protein   (weeks)   Saline   mg/kg   mg/kg   mg/kg   (20 mg/kg)                                                 VLDL   0   0   0   0   0   0           2   0   25   30   40   15           6   0   10   −30   15   −5       LDL   0   0   0   0   0   0           2   0   −30   10   40   10           6   0   −10   55   90   −10       HDL   0   0   0   0   0   0           2   0   5   10   10   15           6   0   10   45   50   20                  
 
     Example 57  
     [0568] Target Validation—Effects of Antisense Inhibition of Apolipoprotein B (ISIS 147764) on Serum AST and ALT Levels  
     [0569] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on liver enzyme (AST and ALT) levels. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks.  
     [0570] AST and ALT levels were measured at 6 weeks and this data is shown in Table 29. Values in the table are expressed as IU/L. Increased levels of the liver enzymes ALT and AST indicate toxicity and liver damage.  
     [0571] Mice treated with ISIS 147764 showed no significant change in AST levels over the duration of the study compared to saline controls (105, 70 and 80 IU/L for doses of 5, 25 and 50 mg/kg, respectively compared to 65 IU/L for saline control). Mice treated with atorvastatin at a daily oral dose of 20 mg/kg had AST levels of 85 IU/L.  
     [0572] ALT levels were increased by all treatments over the duration of the study compared to saline controls (50, 70 and 100 IU/L for doses of 5, 25 and 50 mg/kg, respectively compared to 25 IU/L for saline control). Mice treated with atorvastatin at a daily oral dose of 20 mg/kg had AST levels of 40 IU/L.  
     Example 58  
     [0573] Target Validation—Effects of Antisense Inhibition of Apolipoprotein B (ISIS 147764) on Serum Glucose Levels  
     [0574] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on serum glucose levels. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks.  
     [0575] At study termination, animals were sacrificed 48 hours after the final injections and evaluated for serum glucose levels. ISIS 147764 showed a dose-response effect, reducing serum glucose levels to 225, 190 and 180 mg/dL at doses of 5, 25 and 50 mg/kg, respectively compared to the saline control of 300 mg/dL. Mice treated with atorvastatin at a daily oral dose of 20 mg/kg had serum glucose levels of 215 mg/dL. These data demonstrate that ISIS 147764 is capable of reducing serum glucose levels in high fat fed mice.  
     Example 59  
     [0576] Target Validation—Effects of Antisense Inhibition of Apolipoprotein B (ISIS 147764) on Body, Spleen, Liver and Fat Pad Weight  
     [0577] Male C57BL/6 mice (n=8) receiving a high fat diet (60% kcal fat) were evaluated over the course of 6 weeks for the effects of ISIS 147764 on body, spleen, liver and fat pad weight. Control animals received saline treatment (50 mg/kg). Mice were dosed intraperitoneally every three days (twice a week), after fasting overnight, with 5, 25, 50 mg/kg ISIS 147764 (SEQ ID NO: 645) or saline (50 mg/kg) for six weeks.  
     [0578] At study termination, animals were sacrificed 48 hours after the final injections and body, spleen, liver and fat pad weights were measured. These data are shown in Table 29. Values are expressed as percent change in body weight or organ weight compared to the saline-treated control animals. Data from mice treated with atorvastatin at a daily dose of 20 mg/kg are also shown in the table. Negative values indicated a decrease in weight.  
               TABLE 29                          Effects of Antisense Inhibition of Mouse Apolipoprotein       B on Body and Organ Weight                         Percent Change                             Dose   Atorvastatin                                 Tissue   5 mg/kg   25 mg/kg   50 mg/kg   20 mg/kg                                         Total Body   5   5   −4   1       Wt.       Spleen   10   10   46   10       Liver   18   70   80   15       Fat   10   6   −47   7                  
 
     [0579] These data show a decrease in fat over the dosage range of ISIS 147764 counterbalanced by an increase in both spleen and liver weight with increased dose to give an overall decrease in total body weight.  
     Example 60  
     [0580] Target Validation—Effects of Antisense Inhibition of Apolipoprotein B (ISIS 147764) in b6.129p-apoe tm1unc  Knockout Mice: Lean Animals vs. High Fat-Fed Animals  
     [0581] B6.129P-ApoE tm1Unc  knockout mice (herein referred to as ApoE knockout mice) obtained from The Jackson Laboratory (Bar Harbor, Me.), are homozygous for the Apoe tm1Unc  mutation and show a marked increase in total plasma cholesterol levels that are unaffected by age or sex. These animals present with fatty streaks in the proximal aorta at 3 months of age. These lesions increase with age and progress to lesions with less lipid but more elongated cells, typical of a more advanced stage of pre-atherosclerotic lesion.  
     [0582] The mutation in these mice resides in the apolipoprotein E (ApoE) gene. The primary role of the ApoE protein is to transport cholesterol and triglycerides throughout the body. It stabilizes lipoprotein structure, binds to the low density lipoprotein receptor (LDLR) and related proteins, and is present in a subclass of HDLs, providing them the ability to bind to LDLR. ApoE is expressed most abundantly in the liver and brain.  
     [0583] Female B6.129P-Apoe tm1Unc  knockout mice (ApoE knockout mice) were used in the following studies to evaluate antisense oligonucleotides as potential lipid lowering compounds.  
     [0584] Female ApoE knockout mice ranged in age from 5 to 7 weeks and were placed on a normal diet for 2 weeks before study initiation. ApoE knockout mice were then fed ad libitum a 60% fat diet, with 0.15% added cholesterol to induce dyslipidemia and obesity. Control animals were maintained on a high-fat diet with no added cholesterol. After overnight fasting, mice from each group were dosed intraperitoneally every three days with saline, 50 mg/kg of a control antisense oligonucleotide (ISIS 29837 TCGATCTCCTTTTATGCCCG; SEQ ID NO. 660) or 5, 25 or 50 mg/kg ISIS 147764 (SEQ ID NO: 645) for six weeks.  
     [0585] The control oligonucleotide is a chimeric oligonucleotide (“gapmer”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines.  
     [0586] At study termination and forty eight hours after the final injections, animals were sacrificed and evaluated for target mRNA levels in liver by RT-PCR methods verified by Northern Blot analysis, glucose levels, cholesterol and lipid levels by HPLC separation methods and triglyceride and liver enzyme levels (performed by LabCorp Preclinical Services; San Diego, Calif.). Data from ApoE knockout mice treated with atorvastatin at a daily dose of 20 mg/kg are also shown in the table for comparison.  
     [0587] The results of the comparative studies are shown in Table 29. Data are normalized to saline controls.  
               TABLE 30                          Effects of ISIS 147764 Treatment on Apolipoprotein       B mRNA, Cholesterol, Glucose, Lipid, Triglyceride       and Liver Enzyme Levels in Apoe Knockout Mice.                         Percent Inhibition                             Dose   atorvastatin (20                                                 Control   5 mg/kg   25 mg/kg   50 mg/kg   mg/kg)                                                 mRNA       0   2   42   70   10                         Glucose Levels (mg/dL)                                         Glucose       225   195   209   191   162                         Cholesterol Levels (mg/dL)                                         Cholesterol       1750   1630   1750   1490   938                         Lipoprotein Levels (mg/dL)                                         Lipoprotein   HDL   51   49   62   61   42           LDL   525   475   500   325   250           VLDL   1190   1111   1194   1113   653                         Liver Enzyme Levels (IU/L)                                         Liver Enzymes   AST   55   50   60   85   75           ALT   56   48   59   87   76                  
 
     [0588] It is evident from these data that treatment with ISIS 147764 lowered glucose and cholesterol as well as all lipoproteins investigated (HDL, LDL and VLDL) in ApoE knockout mice. Further, these decreases correlated with a decrease in both protein and RNA levels of apolipoprotein B, demonstrating an antisense mechanism of action. No significant changes in liver enzyme levels were observed, indicating that the antisense oligonucleotide was not toxic to either treatment group.  
     Example 61  
     [0589] Antisense Inhibition of Human BH3 Interacting Domain Death Agonist mRNA Expression by Chimeric Phosphorothioate Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap  
     [0590] The Bcl-2 family of proteins, which includes both positive and negative regulators of apoptosis, act as checkpoints upstream of activated protease cascades orchestrated by caspases and are required for all aspects of cell death. BH3 interacting domain death agonist is a member of the Bcl-2 family and has been shown to dimerize with either Bcl-2, a cell death antagonist, or Bax, a cell death agonist. Due to the integral role played by BH3 interacting domain death agonist in apoptosis, the pharmacological modulation of BH3 interacting domain death agonist activity and/or expression may therefore be an appropriate point of therapeutic intervention in pathological conditions involving deregulated cell death.  
     [0591] Oligonucleotides targeting human BH3 interacting domain death agonist were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0592] Probes and primers to human BH3 Interacting domain death agonist were designed to hybridize to a human BH3 Interacting domain death agonist sequence, using published sequence information (GenBank accession number NM — 001196.1, incorporated herein as SEQ ID NO: 661). For human BH3 Interacting domain death agonist the PCR primers were: forward primer: AGAAGACATCATCCGGAATATTGC (SEQ ID NO: 662) reverse primer: GGAGGGATGCTACGGTCCAT (SEQ ID NO: 663) and the PCR probe was: FAM-AGGCACCTCGCCCAGGTCGG-TAMRA (SEQ ID NO: 664) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0593] The series of oligonucleotides was designed to target different regions of the human BH3 Interacting domain death agonist RNA, using published sequences (GenBank accession number NM — 001196.1, incorporated herein as SEQ ID NO: 661, and residues 12001-28000 of GenBank accession number AC006285, incorporated herein as SEQ ID NO: 665). The oligonucleotides are shown in Table 31. “Target site” indicates the first (5=-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 31 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE)nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on human BH3 Interacting domain death agonist mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments in which T-24 cells were cultured as described in Section 15 (15. Cell Lines for Assaying Oligonucleotide Activity) and treated with oligonucleotides 119845-119922 (SEQ ID NOs: 666-743) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 31                          Inhibition of Human BH3 Interacting Domain Death Agonist mRNA Levels           by Chimeric Phosphorothioate Oligonucleotides       Having 2′-Moe Wings and a Deoxy Gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO                                                     119845   Coding   661   354   ctttcagaatctgcctctat   67   666           119846   Coding   661   707   agtccatcccatttctggct   74   667       119847   5′ UTR   665   60   actgtggtgagtctcccacc   88   668       119848   5′ UTR   665   2083   agtgtcccagtggcgacctg   90   669       119849   Coding   665   2134   cacagtccatggcctgggca   98   670       119850   Intron   665   3582   ctccgcttcctcactccgaa   84   671       119851   Intron   665   3845   tactcgggaggctgaggcag   88   672       119852   Intron   665   3906   ccgtctttactaagatacaa   90   673       119853   Intron   665   4540   tcaagacagtaaatcctgca   93   674       119854   Intron   665   4580   ctttttagatcacaggaaaa   89   675       119855   Intron   665   4987   gccatttaattccaagaata   92   676       119856   Intron   665   5092   ggcccactgagtggacagct   93   677       119857   Intron   665   5373   gcatctgttgtttaaagcca   81   678       119858   Intron   665   5778   acggagcagccgcatggcac   85   679       119859   Intron   665   6999   ggtttcaccatgttggtcag   85   680       119860   Intron   665   7125   tctcggctcactacaacctc   75   681       119861   Intron   665   7369   agggacgctgagatctgcgc   92   682       119862   Intron   665   8083   ggtctcaacaggcagaggca   83   683       119863   Coding   665   8254   atccctgaggctggaaccgt   96   684       119864   Coding   665   8282   caaacaccagtaggtttgtg   92   685       119865   Coding   665   8287   gaagccaaacaccagtaggt   86   686       119866   Coding   665   8318   tgcggaagctgttgtcagaa   81   687       119867   Coding   665   8362   gggagccagcactggcagct   79   688       119868   Coding   665   8418   cgggagtggctgctgcggtt   88   689       119869   Intron   665   9135   gctggacctgggtttcctca   86   690       119870   Intron   665   9353   aagcagccccttggcaaagg   94   691       119871   Intron   665   9424   agggctggatctggaagtgg   74   692       119872   Intron   665   9797   agaaggcagagacattctca   93   693       119873   Intron   665   9875   gcccttcctggaccttccca   95   694       119874   Intron   665   9992   ctcagtctagaggcaaaggc   90   695       119875   Intron   665   10172   ctgatccgtctgtgtccagc   96   696       119876   Intron   665   10643   aagtagctgggattacaggc   83   697       119877   Intron   665   11311   ggccctgtacctagctccca   94   698       119878   Intron   665   11394   atcataccactacactccag   18   699       119879   Intron   665   11641   ttgtattttaagtagagacg   85   700       119880   Intron   665   12649   acaaggccagcccccactgg   74   701       119881   Intron   665   12734   ggcagagacagagcagactc   77   702       119882   Coding   665   12795   tgcctggcaatattccggat   95   703       119883   Coding   665   12811   cccgacctgggcgaggtgcc   99   704       119884   Coding   665   12832   gatgctacggtccatgctgt   97   705       119885   Coding   665   12894   acctcctccgaccggctggt   98   706       119886   Coding   665   14042   ccagggcagtggccaggtcc   95   707       119887   Coding   665   14067   ctagggtaggcctgcagcag   94   708       119888   Coding   665   14072   tgtctctagggtaggcctgc   94   709       119889   Coding   665   14151   cggagcaaggacggcgtgtg   97   710       119890   Coding   665   14178   aaattcactgttgtgtgaaa   96   711       119891   Coding   665   14198   tgcgtaggttctggttaata   98   712       119892   Intron   665   14635   agagcagtgggatcacaggc   80   713       119893   Intron   665   14694   tgttggccagggtggtctgg   77   714       119894   Intron   665   16361   agctgtccatacagactgct   90   715       119895   Coding   665   16678   cttctggaactgtccgttca   96   716       119896   3′ UTR   665   16753   gttgacatgccagggctccg   98   717       119897   3′ UTR   665   16798   atagaagtcacagctatctt   95   718       119898   3′ UTR   665   16933   tgtagatttacagatgtgca   68   719       119899   3′ UTR   665   17176   ttaagatagatagtccctat   89   720       119900   3′ UTR   665   17185   tccttagtattaagatagat   84   721       119901   3′ UTR   665   17236   tagttcagaatctctgtgcc   62   722       119902   3′ UTR   665   17267   ccggacttcccatcatttga   86   723       119903   3′ UTR   665   17293   aaaagtcaagcccctgtgta   77   724       119904   3′ UTR   665   17300   aagttgaaaaagtcaagccc   59   725       119905   3′ UTR   665   17391   gtaaacaaacagtggctgac   82   726       119906   3′ UTR   665   17415   gtatgcagttagttacctga   86   727       119907   3′ UTR   665   17439   tgatgtcatggaaagagaaa   80   728       119908   3′ UTR   665   17452   tttagcaaagtcttgatgtc   72   729       119909   3′ UTR   665   17456   tgtctttagcaaagtcttga   89   730       119910   3′ UTR   665   17588   aacctgttctctccagatgc   80   731       119911   3′ UTR   665   17592   tagaaacctgttctctccag   85   732       119912   3′ UTR   665   17596   tgcttagaaacctgttctct   90   733       119913   3′ UTR   665   17632   aatttttaaaaagtccaact   24   734       119914   3′ UTR   665   17731   tgttgcactgtttctaaagc   85   735       119915   3′ UTR   665   17757   agcttaccactggaacagca   94   736       119916   3′ UTR   665   17764   gggacatagcttaccactgg   70   737       119917   3′ UTR   665   17779   tttaaactgattcctgggac   89   738       119918   3′ UTR   665   17802   gacccagcatccactgtcgt   36   739       119919   3′ UTR   665   17904   gaagaaatcatgagtccgtc   86   740       119920   3′ UTR   665   17942   gattttaaactcttaaagaa   29   741       119921   3′ UTR   665   17966   tagagtttgtttttcctttc   77   742       119922   3′ UTR   665   17970   aatatagagtttgtttttcc   50   743                  
 
     [0594] As shown in Table 31, SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 700, 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, 735, 736, 737, 738, 740, 742 and 743 demonstrated at least 50% inhibition of human BH3 Interacting domain death agonist expression in this assay and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 62  
     [0595] Antisense Inhibition of Mouse BH3 Interacting Domain Death Agonist mRNA Expression by Chimeric Phosphorothioate Oligonucleotides having 2′-Moe Wings and a Deoxy Gap.  
     [0596] Oligonucleotides targeting mouse BH3 Interacting domain Death agonist were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0597] For mouse BH3 Interacting domain Death agonist the PCR primers were: forward primer: TCGAAGACGAGCTGCAGACA (SEQ ID NO: 746) reverse primer: TGGCTCTATTCTTCCTTGGTTGA (SEQ ID NO: 747) and the PCR probe was: FAM-CAGCCAGGCCAGCCGCTCC-TAMRA (SEQ ID NO: 748) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0598] The series of oligonucleotides was designed to target different regions of the mouse BH3 Interacting domain Death agonist using published sequences (GenBank accession number U75506.1, incorporated herein as SEQ ID NO: 744, and residues 9000-120000 of GenBank accession number AC006945, incorporated herein as SEQ ID NO: 745). The oligonucleotides are shown in Table 32. “Target site” indicates the first (5=-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 32 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on mouse BH3 Interacting domain Death agonist mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from two experiments in which b.END cells were cultured and treated with oligonucleotides 119925-120002 (SEQ ID NOs: 749-846) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data.” 
                   TABLE 32                          Inhibition of Mouse BH3 Interacting domain Death agonist mRNA Levels           by Chimeric Phosphorothioate Oligonucleotides       Having 2′-MOE Wings and a Deoxy Gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO                                                     119925   Start Codon   744   21   cgttgctgacctcagagtcc   48   749           119926   Coding   744   232   ctttcagaatctggctctat   32   750       119927   5′ UTR   742   4669   ggcccggcgctctactccac   39   751       119928   5′ UTR   748   4699   gctaaggcaaaggtttgcgg   58   752       119929   5′ UTR   748   5004   cgggtccaccaggaggcctg   42   753       119930   5′ UTR   748   5693   gccatggcaccaggcagtag   71   754       119931   5′ UTR   748   6758   gccaggcagcgtgcccagaa   74   755       119932   5′ UTR   748   7548   cttccccattcatacaccta   61   756       119933   5′ UTR   748   7977   cacttgacaccaacagagac   58   757       119934   5′ UTR   748   8859   gaagcctgtaatcctggcac   73   758       119935   5′ UTR   748   9373   gaccatgtcctggccagaaa   83   759       119936   5′ UTR   748   9439   gtcagtccagtaagggcttt   61   760       119937   5′ UTR   748   9698   ttagcttagccacagaggga   80   761       119938   5′ UTR   748   9768   cgcctgtgctctcttcctgc   53   762       119939   5′ UTR   748   10495   cccatcttctggcctccttg   35   763       119940   5′ UTR   748   11230   ctgaaactccaggctcagga   76   764       119941   5′ UTR   748   12652   ctcatggcagctgcagcagt   66   765       119942   5′ UTR   748   14187   cttgaaaaggaacaaagtgg   44   766       119943   5′ UTR   748   14566   tctatacactactcataacc   55   767       119944   5′ UTR   748   17953   ccatcacagaggccacttct   41   768       119945   5′ UTR   748   18196   tccatccctggaacaatgtg   58   769       119946   5′ UTR   748   19488   cagagctcagctttcttccc   68   770       119947   5′ UTR   748   19741   agctcacagagtccagggaa   55   771       119948   5′ UTR   748   19752   caagcactgccagctcacag   59   772       119949   Coding   748   19782   tcagagtccatggcacaagc   61   773       119950   Intron   748   20989   ttgccaaacagaagacacca   3   774       119951   Intron   748   21013   gcagagaaacaggctgtggt   42   775       119952   Coding   748   21182   gtctgtgatgtgcttggccc   63   776       119953   Coding   748   21205   tggagaaagccgaacaccag   57   777       119954   Coding   748   21259   acaggcagttcccgacccag   71   778       119955   Coding   748   21282   ggtctgcctcccagtaagct   27   779       119956   Coding   748   21306   cgtctgtctgcagctcgtct   89   780       119957   Intron   748   21950   cttttctgaatgacttgata   39   781       119958   Intron   748   22293   cactgataggaagtgtgtcc   54   782       119959   Intron   748   22835   ctcagttgctgtaaacacag   57   783       119960   Intron   748   22883   ccacagcgctctgagcactc   73   784       119961   Intron   748   23125   gtcctgaagtatcctgacct   72   785       119962   Intron   748   23239   gaaataaactagccagaggg   26   786       119963   Coding   748   24169   tttcttcctgactttcagaa   33   787       119964   Coding   748   24201   ttgggcgagatgtctggcaa   55   788       119965   Coding   748   24208   cgcctatttgggcgagatgt   51   789       119966   Coding   748   24264   gaactgtgcggctagctgtc   62   790       119967   Intron   748   24515   cgccacaagagaagactgag   54   791       119968   Intron   748   24877   aatgtgtgtgtctttgacag   53   792       119969   Intron   748   25363   ctacatgttatcttcccttc   37   793       119970   Coding   748   25705   agggctttggccaggcagtt   43   794       119971   Coding   748   25776   acagcattgtcattatcagc   67   795       119972   Coding   748   25814   gagcaaagatggtgcgtgac   54   796       119973   Coding   748   25830   tgtggaagacatcacggagc   78   797       119974   Coding   748   25838   gacagtcgtgtggaagacat   48   798       119975   Coding   748   25858   aggttctggttaataaagtt   34   799       119976   Intron   748   26838   gtcattttccagcagtctca   77   800       119977   Coding   748   27236   gcgggctcctcagtccatct   74   801       119978   3′ UTR   748   27315   gttctctgggacctgtcttc   44   802       119979   3′ UTR   748   27474   tcattcccaagtgggaaccc   49   803       119980   3′ UTR   748   27577   cagaagcccacctacatggt   44   804       119981   3′ UTR   748   27608   atgcacctctcctaatgctg   58   805       119982   3′ UTR   748   27612   gccgatgcacctctcctaat   67   806       119983   3′ UTR   748   27657   gagcacttcagtgtccacta   56   807       119984   3′ UTR   748   27700   agatcagccattcggctttt   58   808       119985   3′ UTR   748   27711   cccatggtttgagatcagcc   75   809       119986   3′ UTR   748   27788   gatagaaatcttgagataat   11   810       119987   3′ UTR   748   27834   caccacacagataagtcgtg   65   811       119988   3′ UTR   748   27842   gtaactgacaccacacagat   60   812       119989   3′ UTR   748   27851   agcctgagtgtaactgacac   54   813       119990   3′ UTR   748   27859   gtagcaagagcctgagtgta   48   814       119991   3′ UTR   748   27868   ttgcattccgtagcaagagc   51   815       119992   3′ UTR   748   27934   agtgacctgctgctgtttta   37   816       119993   3′ UTR   748   28042   cttttgatatggaatcttct   50   817       119994   3′ UTR   748   28067   aatacagaagcggagggaac   32   818       119995   3′ UTR   748   28083   gaggccttgtctctgaaata   78   819       119996   3′ UTR   748   28107   cgtaacaacgcttgaggata   63   820       119997   3′ UTR   748   28145   gctgacgatcccagctttaa   38   821       119998   3′ UTR   748   28167   cttgcaggctgtggcggctc   65   822       119999   3′ UTR   748   28170   atacttgcaggctgtggcgg   52   823       120000   3′ UTR   748   28192   ctgggatgagttcagaacta   73   824       120001   3′ UTR   748   28332   cacatatttttagaacagaa   38   825       120002   3′ UTR   748   28378   gagccttttattttgaagaa   60   826                  
 
     [0599] As shown in Table 32, SEQ ID NOs 749, 750, 751, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762, 763, 764,765, 766, 767, 768, 769, 770, 771, 772, 773, 775, 776, 777, 778, 780, 781, 782, 783, 784, 785, 787, 788, 789, 790, 791, 792, 793, 794, 795, 796, 797, 798, 799, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 811, 812, 813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825 and 826 demonstrated at least 30% inhibition of mouse BH3 Interacting domain death agonist expression in this experiment and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 63  
     [0600] Target Validation—Effect of BH3 Interacting Death Domain Antisense Oligonucleotides in a Fas Cross-Linking Antibody Murine Model for Hepatitis  
     [0601] Injection of agonistic Fas-specific antibody into mice can induce massive hepatocyte apoptosis and liver hemorrhage, and death from acute hepatic failure (Ogasawara, J., et al.,  Nature,  1993, 364, 806-809). Apoptosis-mediated aberrant cell death has been shown to play an important role in a number of human diseases. For example, in hepatitis, Fas and Fas ligand up-regulated expression are correlated with liver damage and apoptosis. It is thought that apoptosis in the livers of patients with fulminant hepatitis, acute and chronic viral hepatitis or autoimmune hepatitis, as well as chemical or drug induced liver intoxication may result from Fas activation on hepatocytes. There are various indices of liver damage and/or apoptosis that are commonly used. These include measurement of the liver enzymes, AST and ALT.  
     [0602] Eight to ten week-old female Balb/c mice were intraperitoneally injected with oligonucleotide 119935 (SEQ ID NO. 759) at 24 mg/kg, daily for 4 days or with saline at a dose of 7 ug. Four hours after the last dose, 7.5 ug of mouse Fas antibody (Pharmingen, San Diego, Calif.) was injected into the mice. Mortality of the mice was measured for 48 hours following antibody treatment. Results are shown in Table 33. Mortality is expressed as percent survival.  
               TABLE 33                          Protective Effects of BH3 Interacting Death Domain       Antisense Chimeric (deoxy gapped) Phosphorothioate       Oligonucleotides in Fas Antibody Cross-linking Induced       Death in Balb/c Mice                             SEQ ID   Percent Survival                                             ISIS #   NO:   4 Hr   6 Hr   8 Hr   12 Hr   24 Hr   48                                                     Saline   —   100   90   20   0   0   0       119935   107   100   100   100   100   100   100                  
 
     [0603] Oligonucleotide 119935 (SEQ ID NO. 759) completely protected the Fas-antibody treated mice from death. Injection with saline alone did not confer any protective effect.  
     [0604] After challenge with a higher dose of Fas antibody (15 ug), protection from fulminant death by the BH3 interacting death domain antisense oligonucleotides was lost with survival rates dropping to 1 percent at 5 hours post-treatment. An increase in antisense oligonucleotide dosage to 50 mg/kg given 6 times every 3 days also failed to produce protection from fulminant death at the higher dose of Fas antibody.  
     [0605] The BH3 interacting death domain antisense oligonucleotide was also shown to override sensitization to Fas antibody-induced death by Bcl-xL antisense oligonucleotide in the same model.  
     [0606] In these experiments, 8-10 week-old female Balb/c mice were intraperitoneally injected with oligonucleotides ISIS 16009 (SEQ ID NO. 827, targeting murine Bcl-xL) alone or in combination with ISIS 119935 (SEQ ID NO. 756) at 50 mg/kg, 6 times a day for two days or with saline at a dose of 7 ug. ISIS 16009 is a chimeric oligonucleotide (“gapmer”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. Cytidine residues in the “wings” are 5-methylcytidines. Four hours after the last dose, 7 ug of mouse Fas antibody (Pharmingen, San Diego, Calif.) was injected into the mice. Mortality of the mice was measured for 48 hours following antibody treatment. Results are shown in Table 34. Mortality is expressed as percent survival. N.D. indicates no data for these timepoints.  
               TABLE 34                          Protective Effects of BH3 Interacting Death       Domain Antisense Oligonucleotides in       Fas Antibody Cross-linking Induced Death in       Balb/c Mice sensitized by Bcl-xL       antisense oligonucleotide treatment.                         Percent Survival                                             ISIS #   SEQ ID   4 Hr   6 Hr   8 Hr   12 Hr   24 Hr   48 Hr                                                     saline   —   90   60   20   0    0    0       16009   175   90   30   20   10   N.D.   N.D.       119935 +   107   100   100   100   100   100   100       16009                  
 
     [0607] Western blot analysis of Bcl-xL and BH3 interacting death domain proteins revealed that the expression levels of both targets was reduced to greater than 90%.  
     Example 64  
     [0608] Target Validation—Effect of BH3 Interacting Death Domain Antisense Oligonucleotides in an Endotoxin and D(+)-Galactosamine-induced Murine Model of Fulminant Hepatitis and Liver Injury  
     [0609] The lipopolysaccharide/D-galactosamine or LPS/GalN model is a well known experimental model of toxin-induced hepatitis. Injection of the endotoxin, lipopolysaccharide (LPS), induces septic shock death in the mouse, though with LPS alone, the mouse liver does not sustain major damage. Injection of D-Galactosamine (GalN), while metabolized in liver causing depletion of UTP, is not lethal to mice. It does, however, sensitize animals to TNF-α or LPS-induced endotoxic shock by over 1,000 fold. In the presence of GalN, LPS induces apoptotic cell death in liver, thymus, spleen, lymph nodes and the kidney and results in fulminant death in animals. The liver injury is known to be transferable via the serum, suggesting a mechanism of action under TNF-α control. Further support for this mechanism is provided by the finding that TNFR1 knockout mice are resistant to LPS/GalN-induced liver injury and death.  
     [0610] Eight-week-old female Balb/c mice were used to assess the activity of BH3 interacting death domain antisense oligonucleotides in the endotoxin and D(+)-Galactosamine-induced murine model of fulminant hepatitis and liver injury. Mice were intraperitoneally pretreated with 24 mg/kg of ISIS 119935 (SEQ ID NO. 759) four times a day for 2 days. Control mice were injected with saline. One day after the last dose of oligonucleotide, mice were injected intraperitoneally with 5 ng LPS (DIFCO laboratories) and 20 mg D-Galactosamine (Sigma) per animal in saline. At time intervals of 5.5, 7.5, 9.5, 21.5, 30, 45 and 53 hours after the final dose, animals were monitored for survival rates. Results are shown in Table 35.  
               TABLE 35                          Protective Effects of BH3 Interacting Death       Domain Antisense Oligonucleotides in       Endotoxin-Mediated Death in Balb/c Mice                         Percent Survival                                                         5.5   7.5                           ISIS #   SEQ ID   Hr   Hr   9.5 Hr   21.5 Hr   30 Hr   45 Hr   53 Hr                                                         Saline   —   100   100   20   20   10   10   10       119935   107   100   100   100   100   100   100   100                  
 
     [0611] BH3 interacting death domain antisense oligonucleotides were also shown to override sensitization to endotoxin-mediated death by Bcl-xL antisense oligonucleotides in the same model. In these experiments, 8-10 week old female Balb/c mice were intraperitoneally pretreated with 24 mg/kg of ISIS 16009 (SEQ ID NO. 827) alone or in combination with ISIS 119935 (SEQ ID NO. 756) four times a day for 2 days. Control mice were injected with saline. One day after the last dose of oligonucleotide, mice were injected intraperitoneally with 5 ng LPS (DIFCO laboratories) and 20 mg D-Galactosamine (Sigma) per animal in saline. At time intervals of 6, 6.5, 7, 7.5, 9, 9.5 and 22 hours after the final dose, animals were monitored for survival rates. Results are shown in Table 36. Mortality is expressed as percent survival.  
               TABLE 36                          Protective Effects of BH3 Interacting Death       Domain Antisense Oligonucleotides in       Endotoxin-Mediated Death in Balb/c Mice       sensitized by Bcl-xL antisense       oligonucleotide treatment.       Percent Survival                                                     SEQ                       9.5           ISIS #   ID   6 Hr   6.5 Hr   7 Hr   7.5 Hr   9 Hr   Hr   22 Hr                                                         Saline   —   100   100   100   100   70   20   10       16009   175   100   80   30   0   0   0   0       119935 +   107   100   100   100   100   100   100   100       16009                  
 
     Example 65  
     [0612] Antisense Inhibition of PTEN mRNA Expression by Chimeric Phosphorothioate Oligonucleotides Having 2′-Moe Wings and a Deoxy Gap  
     [0613] PTEN is a dual-specificity protein phosphatase recently implicated as a phosphoinositide phosphatase in the insulin-signaling pathway. The pharmacological modulation of PTEN activity and/or expression may be an appropriate point for therapeutic intervention in metabolic disorders such as diabetes which arise from degregulated insulin signaling.  
     [0614] Oligonucleotides targeting human PTEN were designed as described in Example 2, synthesized as described in Examples 3-7, analyzed as described in Example 8 and assayed by RT-PCR as described in Example 12.  
     [0615] Probes and primers to human PTEN were designed to hybridize to a human PTEN sequence, using published sequence information (GenBank accession number U92436.1, incorporated herein as SEQ ID NO: 828). For human PTEN the PCR primers were: forward primer: AATGGCTAAGTGAAGATGACAATCAT (SEQ ID NO: 829) reverse primer: TGCACATATCATTACACCAGTTCGT (SEQ ID NO: 830) and the PCR probe was: FAM-TTGCAGCAATTCACTGTAAAGCTGGAAAGG-TAMRA (SEQ ID NO: 831) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0616] The series of oligonucleotides was designed to target different regions of the human PTEN RNA, using published sequences (GenBank accession number U92436.1, incorporated herein as SEQ ID NO: 828). The oligonucleotides are shown in Table 37. “Target site” indicates the first (5=-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. All compounds in Table 37 are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotide. All cytidine residues are 5-methylcytidines. The compounds were analyzed for their effect on human PTEN mRNA levels by quantitative real-time PCR as described in other examples herein. Data are averages from three experiments in which T-24 cells were cultured as described in Section 15 (15. Cell Lines for Assaying Oligonucleotide Activity) treated with oligonucleotides 29574-29613 (SEQ ID NOs: 832-867) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 37                          Inhibition of Human PTEN mRNA levels           by chimeric phosphorothioate oligonucleotides       having 2′-MOE wings and a deoxy gap                                                     TARGET                           ISIS #   REGION   SEQ ID NO   TARGET SITE   SEQUENCE   % INHIB   SEQ ID NO.                                                     29574   5′ UTR   828   19   cgagaggcggacgggacc   29   832           29575   5′ UTR   828   57   cgggcgcctcggaagacc   27   833       29576   5′ UTR   828   197   tggctgcagcttccgaga   67   834       29577   5′ UTR   828   314   cccgcggctgctcacagg   79   835       29578   5′ UTR   828   421   caggagaagccgaggaag   51   836       29579   5′ UTR   828   494   gggaggtgccgccgccgc   71   837       29581   5′ UTR   828   671   ccgggtccctggatgtgc   88   838       29582   5′ UTR   828   757   cctccgaacggctgcctc   66   839       29583   5′ UTR   828   817   tctcctcagcagccagag   74   840       29584   5′ UTR   828   891   cgcttggctctggaccgc   81   841       29585   5′ UTR   828   952   tcttctgcaggatggaaa   63   842       29587   Coding   828   1106   ggataaatataggtcaag   50   843       29588   Coding   828   1169   tcaatattgttcctgtat   41   844       29589   Coding   828   1262   ttaaatttggcggtgtca   74   845       29590   Coding   828   1342   caagatcttcacaaaagg   64   846       29591   Coding   828   1418   attacaccagttcgtccc   55   847       29592   Coding   828   1504   tgtctctggtccttactt   64   848       29593   Coding   828   1541   acatagcgcctctgactg   73   849       29595   Coding   828   1694   gaatatatcttcaccttt   30   850       29596   Coding   828   1792   ggaagaactctactttga   61   851       29597   Coding   828   1855   tgaagaatgtatttaccc   31   852       29599   Coding   828   2020   ggttggctttgtctttat   56   853       29600   Coding   828   2098   tgctagcctctggatttg   80   854       29601   Coding   828   2180   tctggatcagagtcagtg   65   855       29602   3′ UTR   828   2268   tattttcatggtgtttta   41   856       29603   3′ UTR   828   2347   tgttcctataactggtaa   63   857       29604   3′ UTR   828   2403   gtgtcaaaaccctgtgga   39   858       29605   3′ UTR   828   2523   actggaataaaacgggaa   6   859       29606   3′ UTR   828   2598   acttcagttggtgacaga   40   860       29607   3′ UTR   828   2703   tagcaaaacctttcggaa   22   861       29608   3′ UTR   828   2765   aattatttcctttctgag   23   862       29609   3′ UTR   828   2806   taaatagctggagatggt   27   863       29610   3′ UTR   828   2844   cagattaataactgtagc   42   864       29611   3′ UTR   828   2950   ccccaatacagattcact   10   865       29612   3′ UTR   828   3037   attgttgctgtgtttctt   61   866       29613   3′ UTR   828   3088   tgtttcaagcccattctt   55   867                  
 
     [0617] As shown in Table 37, SEQ ID NOs: 834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 844, 845, 846, 847, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 860, 864, 866 and 867 demonstrated at least 30% inhibition of PTEN expression in this experiment and are therefore preferred. The target sites to which these preferred sequences are complementary are herein referred to as “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     Example 66  
     [0618] Target Validation—Inhibition of PTEN Expression-Dose Response in Human, Mouse and Rat Hepatocytes  
     [0619] In accordance with the present invention, two additional oligonucleotides targeted to human PTEN were designed and synthesized. ISIS 116847 (CTGCTAGCCTCTGGATTTGA, SEQ ID NO: 868) and ISIS 116845 (ACATAGCGCCTCTGACTGGG, SEQ ID NO: 869). The mismatch control for ISIS 116847 is ISIS 116848 (CTTCTGGCATCCGGTTTAGA, SEQ ID NO: 870), a six base pair mismatch of ISIS 116847, while the control used is the mixed sequence 20-mer negative oligonucleotide control ISIS 29848 (SEQ ID NO: 461). Both ISIS 116847 and ISIS 116845 target the coding region of Genbank accession no. U92436.1 (SEQ ID NO: 828), with ISIS 116847 starting at position 2097 and ISIS 116845 starting at position 1539.  
     [0620] These oligonucleotide sequences also target the mouse PTEN sequence with perfect complementarity, with ISIS 116845 targeting nucleotides 1453-1472 and ISIS 116847 targeting nucleotides 2012-2031 of GenBank accession number U92437 (SEQ ID NO: 871) (locus name MMU92437; Steck et al., Nature Genet., 1997, 15,356-362). For mouse PTEN the PCR primers were: forward primer:  
     [0621] ATGACAATCATGTTGCAGCAATTC (SEQ ID NO: 872) reverse primer:  
     [0622] CGATGCAATAAATATGCACAAATCA (SEQ ID NO: 873) and the PCR probe was:  
     [0623] FAM-CTGTAAAGCTGGAAAGGGACGGACTGGT-TAMRA (SEQ ID NO: 874) where FAM (PE-Applied Biosystems, Foster City, Calif.) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, Calif.) is the quencher dye.  
     [0624] Similarly, these oligonucleotide sequences target the rat PTEN sequence with perfect complementarity, with ISIS 116845 targeting nucleotides 505-524 and ISIS 116847 targeting nucleotides 1063-1082 of GenBank accession number AF017185 (SEQ ID NO: 875). The mouse PTEN primers and probe listed above target the rat PTEN sequence with perfect complementarity and were used to determine the PTEN expression dose response in rat hepatocytes.  
     [0625] All compounds of this example are chimeric oligonucleotides (“gapmers”) 20 nucleotides in length, composed of a central “gap” region consisting of ten 2′-deoxynucleotides, which is flanked on both sides (5′ and 3′ directions) by five-nucleotide “wings”. The wings are composed of 2′-methoxyethyl (2′-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P═S) throughout the oligonucleotides. All cytidine residues are 5-methylcytidines.  
     [0626] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from two experiments.  
     [0627] In a dose-response experiment, human hepatocyte cells (HEPG2; American Type Culture Collection, Manassas, Va.), mouse primary hepatocytes, and rat primary hepatocytes were treated with ISIS 116847 and its mismatch control, ISIS 116848 at doses of 10, 50, 100 and 200 nM oligonucleotide normalized to untreated controls. In all three species, the dose response was linear compared to vehicle treated controls.  
     [0628] In human HEPG2 cells, ISIS 116847 reduced PTEN mRNA levels to 55% of control at a dose of 10 nM, and to 5% of control at 200 nM while the PTEN mRNA levels in cells treated with the mismatch control oligonucleotide remained at greater than 90% of control across the entire dosing range.  
     [0629] In mouse primary hepatocytes the trend was the same with ISIS 116847 reducing PTEN mRNA levels to 85% of control at the lower dose of 10 nM, and down to 2% of control at the 200 nM dose. Again, the control oligonucleotide, ISIS 116848 failed to reduce PTEN mRNA levels and remained at or above 85% of control.  
     [0630] In rat primary hepatocytes, ISIS 116847 reduced PTEN mRNA levels to 55% of control at the lower dose of 10 nM and to 10% of control at the highest dose of 200 nM. PTEN mRNA levels in cells treated with the control oligonucleotide, ISIS 116848, remained at or above 95% of control across the entire dosing range.  
     Example 67  
     [0631] Effects of Inhibition of PTEN on mRNA Expression in Fat and Liver  
     [0632] In the following examples, modulators of PTEN were tested in db/db mice (Jackson Laboratories, Bar Harbor, Me.). These mice are hyperglycemic, obese, hyperlipidemic, and insulin resistant, and are used as a standard animal model of diabetes.  
     [0633] Male db/db mice (age 14 weeks) were divided into matched groups (n=5) with the same average blood glucose levels and treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Wild type mice were similarly treated. Controls included saline, ISIS 116848 (a mismatch control), ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) and the sense control of ISIS 116847. As a comparison db/db mice were also treated with troglitazone, an oral antihyperglycemic agent which is used in the treatment of type II diabetes. It acts primarily to decrease insulin resistance, improve sensitivity to insulin in muscle and adipose tissue and inhibit hepatic gluconeogenesis. At day 28 mice were sacrificed and PTEN mRNA levels were measured.  
     [0634] Treatment of db/db mice with ISIS 116847 showed a dose-dependent decrease in PTEN mRNA levels in the liver to 10% of control at 50 mg/kg. ISIS 116845 showed a reduction in PTEN mRNA levels to 22% of control at a dose of 50 mg/kg.  
     [0635] In wild-type mice a level of 5% of control PTEN mRNA required a dose of 100 mg/kg of ISIS 116847. Neither troglitazone nor any of the controls had an effect on PTEN mRNA levels over saline control.  
     [0636] Similar results were seen in fat. Treatment of db/db mice with ISIS 116847 showed a dose-dependent decrease in PTEN mRNA levels in fat to 20% of control at 50 mg/kg. ISIS 116845 showed a reduction in PTEN mRNA levels to 35% of control at a dose of 50 mg/kg.  
     [0637] In wild-type mice a level of 18% of control required a dose of 100 mg/kg of ISIS 116847. Neither troglitazone nor any of the controls had an effect on PTEN mRNA levels over saline control.  
     [0638] In another experiment, male db/db mice (age 14 weeks) were divided into matched groups (n=5) with the same average blood glucose levels and treated intraperitoneally with saline or ISIS 116847 every other day (q2d) or twice a week (q4d) at a dose of 20 mg/kg. The control for both protocols was the mismatch control, ISIS 116848. Mice were exsanguinated on day 14 and PTEN mRNA levels in liver and fat were measured.  
     [0639] ISIS 116847 successfully reduced PTEN mRNA levels in both liver and fat of db/db mice at both the q2d and q4d dosing schedules in a dose-dependent manner, whereas the mismatch control and saline treated animals showed no reduction in PTEN mRNA.  
     [0640] There was no reduction of PTEN mRNA in skeletal muscle with any of the oligonucleotides used. This lack of an effect in muscle indicates that reduction of expression of PTEN in liver and fat alone is sufficient to lower hyperglycemia.  
     Example 68  
     [0641] Target Validation—Effects of inhibition of PTEN on mRNA Expression in Kidney  
     [0642] Male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Controls included saline, ISIS 116848 (a mismatch control), ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) and the sense control of ISIS 116847. As a comparison db/db mice were also treated with troglitazone. At day 28 mice were sacrificed and PTEN mRNA levels were measured.  
     [0643] Treatment with ISIS 116847 showed a dose-dependent decrease in PTEN mRNA levels in kidney, being reduced to 70% of control at a dose of 50 mg/kg. ISIS 116845 reduced PTEN mRNA levels to 85% of control at the same dose.  
     [0644] In wild-type mice a level of 75% of control required a dose of 100 mg/kg of ISIS 116847. Neither troglitazone nor any of the controls had an effect on PTEN mRNA levels over saline control.  
     Example 69  
     [0645] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on PTEN Protein Levels in Liver Extracts as a Function of Time and Dose  
     [0646] Male db/db and wild-type mice (age 14 weeks) were treated once a week for 4 weeks with saline, a control oligonucleotide, ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) (50 mg/kg) or ISIS 116847 at 10, 25 or 50 mg/kg. Wild-type mice were treated with saline or ISIS 116847 at 100 mg/kg. Mice were sacrificed at day 28 and PTEN protein levels were measured by Western blotting as described in other examples herein.  
     [0647] In the db/db mice, treatment with ISIS 116847 caused a dose-dependent decrease in PTEN protein levels compared to saline controls or mismatch treated animals.  
     [0648] Protein levels in wild-type mice treated at 100 mg/kg were comparably reduced to the levels seen in db/db mice treated at the 50 mg/kg dose. There was no significant difference in the relative levels of PTEN protein in control lean versus db/db mice.  
     Example 70  
     [0649] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on PTEN Protein Levels in Fat and Kidney as a Function of Time and Dose  
     [0650] Male db/db and wild-type mice (age 14 weeks) were treated once a week for 4 weeks with saline or ISIS 116847 at 50 mg/kg by intraperitoneal injection. Mice were sacrificed at day 28 and PTEN protein levels were measured by Western blotting described in other examples herein.  
     [0651] PTEN levels in fat were reduced in both db/db and wild-type mice by the PTEN oligomeric compounds as compared to control, and slight reduction of PTEN levels was seen in the kidney after treatment with oligomeric compounds.  
     Example 71  
     [0652] Target Validation—Effects of Inhibition of PTEN on Blood Glucose Levels  
     [0653] Male db/db and wild type mice (age 14 weeks) were divided into matched groups (n=5) with the same average blood glucose levels and treated by intraperitoneal injection with saline or ISIS 116847 every other day (q2d) or twice a week (q4d) at a dose of 20 mg/kg. The control for both protocols was the mismatch control, ISIS 116848. Blood glucose levels were measured on day 7 and day 14.  
     [0654] By day 14 in db/db mice, blood glucose levels were reduced for both treatment schedules; from starting levels of 330 mg/dL to 175 mg/dL (q2d) and 170 mg/dL (q4d) which are levels within the range considered normal for wild-type mice. The mismatch control levels remained at 310 mg/dL throughout the study.  
     [0655] In wild-type mice, blood glucose levels remained constant throughout the study for all treatment groups (average 115 mg/dL).  
     [0656] In a similar experiment, male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 or ISIS 116845 at 50 mg/kg. Controls included saline, ISIS 116848 (a mismatch control) and ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461). At day 28 mice were sacrificed and serum glucose levels were measured.  
     [0657] In db/db mice, treatment with either ISIS 116847 or ISIS 116845 reduced serum glucose levels relative to saline control (480 mg/dL) to 240 and 280 mg/dL, respectively. This reduction was statistically significant (p&lt;0.005). Neither the mismatch nor universal control had any effect on serum glucose levels. In wild-type animals, ISIS 116847 failed to reduce serum glucose levels from that of control (200 mg/dL).  
     Example 72  
     [0658] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on Blood Glucose Levels of db/db Mice as a Function of Time and Dose  
     [0659] Male db/db mice (age 14 weeks) were treated once a week for 4 weeks with saline or ISIS 116847 at 10, 25 or 50 mg/kg intraperitoneally. Blood glucose levels were measured on day 7, 14, 21 and 28.  
     [0660] At the beginning of the study, all groups had blood glucose levels of 275 mg/dL which rose in the saline treated animals and those treated at the low dose of ISIS 116847 to 350 mg/dL and 320 mg/dL, respectively by day four. At the end of the first week, all three dosing schedules showed a reduction in blood glucose and continued to show linear dose response decreases throughout the study. At day 28, blood glucose levels in animals treated with oligomeric compounds were 275 mg/dL (10 mg/kg dose), 175 mg/dL (25 mg/kg dose) and 120 mg/dL (50 mg/kg dose) while saline treated levels remained at 350 mg/dL. The average glucose levels for oligonucleotide treated mice at the end of the four week study was 194 mg/dL as compared to 418 mg/dL for saline treated controls (p&lt;0.0001).  
     Example 73  
     [0661] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on Blood Glucose Levels of db/db Mice-Insulin Tolerance Test  
     [0662] Male db/db mice (age 14 weeks) were treated once with saline or ISIS 116847 50 mg/kg by intraperitoneal injection. The insulin tolerance test was performed after a four hour fast followed by an intraperitoneal injection of 1 U/kg human insulin (Lilly). On day 21, blood was withdrawn from the tail at 0, 30, 60 and 90 minutes and blood glucose levels were measured as a percentage of blood glucose at time zero. Statistical analysis was performed using ANOVA repeated measures followed by Bonferroni Dunn analysis, p&lt;0.05.  
     [0663] Treatment with ISIS 116847 on day 21 resulted in a significant dose-dependent decrease in blood glucose (p&lt;0.006) at the 90 minute post-treatment time point to 45% of control (55% decrease). Saline treatment resulted in a 30% reduction. These studies indicate that the PTEN oligonucleotide is capable of increasing sensitivity to insulin (decreasing insulin resistance) and that treatment does not cause hypoglycemia. Glucose levels in PTEN treated mice (both db/db and wild-type) fasted for 16 hours remained normal.  
     Example 74  
     [0664] Target Validation—Effects of Inhibition of PTEN on Serum Triglyceride and Cholesterol Concentration  
     [0665] Male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Controls included saline, ISIS 116848 (a mismatch control), ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) and the sense control of ISIS 116847. As a comparison db/db mice were also treated with troglitazone. At day 28 mice were sacrificed and triglyceride and cholesterol levels were measured.  
     [0666] Treatment of db/db mice with ISIS 116847 resulted in a dose-dependent reduction of both triglycerides and cholesterol compared to saline controls (a reduction from 200 mg/dL to 100 mg/dL for triglycerides and from 130 mg/dL to 98 mg/dL for cholesterol). Treatment of db/db mice with ISIS 116845 at a dose of 50 mg/kg resulted in a decrease in both triglycerides and cholesterol levels to 130 mg/dL and 75 mg/dL, respectively. Troglitazone treatment of db/db mice reduced both triglyceride and cholesterol levels to 85 mg/dL each.  
     [0667] Wild-type animals did not respond to treatment with ISIS 116847 at a dose of 100 mg/kg as both triglyceride and cholesterol levels remained similar to control saline treated animals (between 85 and 105 mg/dL). The reductions seen in cholesterol and triglycerides were statistically significant at p&lt;0.005.  
     Example 75  
     [0668] Target Validation—Effects of Inhibition of PTEN on Body Weight  
     [0669] Male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Controls included saline, ISIS 116848 (a mismatch control), ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) and the sense control of ISIS 116847. As a comparison db/db mice were also treated with troglitazone. At day 28 mice were sacrificed and final body weights were measured.  
     [0670] Treatment with ISIS 116847 resulted in a dose-dependent increase in body weight over the dose range with animals treated at the high dose gaining an average of 8.7 grams while saline treated controls gained 2.8 grams. Animals treated with the mismatch or universal control oligonucleotide gained between 2.5 and 3.5 grams of body weight and troglitazone treated animals gained 5.0 grams.  
     [0671] Wild-type animals treated with 100 mg/kg of ISIS 116847 gained 2.0 grams of body weight compared to a gain of 1.3 grams for the wild-type saline or mismatch controls.  
     [0672] Weight gain in the PTEN oligomeric compound treated mice began to increase relative to that in saline or control groups at the same time that glucose levels began to drop.  
     Example 76  
     [0673] Target Validation—Effects of Inhibition of PTEN on Liver Weight-Anterior Lobe  
     [0674] Male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Controls included saline, ISIS 116848 (a mismatch control), ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) and the sense control of ISIS 116847. As a comparison db/db mice were also treated with troglitazone. At day 28 mice were sacrificed and the weights of the anterior lobe of the liver were measured.  
     [0675] db/db animals treated at the high dose had liver weights of 1.2 grams while saline treated controls weighed 0.75 grams. db/db animals treated with ISIS 116845 at a dose of 50 mg/kg had comparable liver size to those treated with ISIS 116847 at a dose of 25 mg/kg (1.0 grams). Animals treated with the mismatch control, universal control or troglitazone had livers weighing an average of 1.0 gram.  
     [0676] Wild-type mouse livers treated with 100 mg/kg of ISIS 116847 weighed 0.7 grams compared to 0.5 grams for the wild-type saline treated controls.  
     [0677] BrdU (bromine deoxyuridine) staining of liver sections indicated that the increase in liver weight was not due to increased cell proliferation, and there was no increase in inflammatory infiltrates in the liver. Long-term studies show that the increases in liver weight are reversed.  
     Example 77  
     [0678] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on PEPCK mRNA Expression in Liver of db/db Mice  
     [0679] PEPCK is the rate-limiting enzyme of gluconeogenesis and is expressed predominantly in liver where it acts in the gluconeogenic pathway (production of glucose from amino acids) and in kidney where it acts in the gluconeogenic pathway as well as being glyceroneogenic and ammoniagenic. In the liver, PEPCK is negatively regulated by insulin and has therefore been considered a potential contributing factor to hyperglycemia in diabetics (Sutherland et al., Philos. Trans. R. Soc. Lond. B. Biol. Sci., 1996, 351, 191-199).  
     [0680] Male db/db mice (age 14 weeks) with the same average blood glucose levels were divided into groups (n=5) and treated intraperitoneally with saline, ISIS 116847 or the mismatch control, ISIS 116848, every other day (q2d). Mice were exsanguinated on day 14 and PEPCK mRNA levels in liver were measured.  
     [0681] Mice treated with ISIS 116847 showed a reduction of PEPCK mRNA to 65% of saline treated controls. The mismatch control group remained at 98% of saline treated control.  
     Example 78  
     [0682] Target Validation—Effects of Inhibition of PTEN (ISIS 116847) on Serum Insulin Levels of db/db Mice  
     [0683] Male db/db and wild type mice (age 14 weeks) were divided into matched groups (n=5) with the same average blood glucose levels and treated by intraperitoneal injection with saline or ISIS 116847 every other day (q2d) or twice a week (q4d) at a dose of 20 mg/kg. The control for both protocols was the mismatch control, ISIS 116848. Mice were exsanguinated on day 14 and serum insulin levels were measured.  
     [0684] On day 14 db/db mice treated on the q2d schedule had serum insulin levels of 7.8 ng/mL, compared to saline treated (9 ng/mL) and mismatch treated animals (12 ng/mL). In the q4d schedule there was a drop in the serum insulin levels of db/db mice treated with ISIS 116847 to 4 ng/mL while the mismatch control levels remained at 12 ng/mL. Wild-type mice had serum insulin levels of 1 ng/mL throughout the course of both treatment schedules.  
     Example 79  
     [0685] Target Validation—Effects of Inhibition of PTEN on Liver Function-AST/ALT Levels  
     [0686] Male db/db and wild type mice (age 14 weeks) were divided into matched groups (n=5) with the same average blood glucose levels and treated by intraperitoneal injection with saline, troglitazone, or ISIS 116847 every other day (q2d) or twice a week (q4d) at a dose of 20 mg/kg. The control for both protocols was the mismatch control, ISIS 116848. Mice were exsanguinated on day 14 and liver enzyme levels were measured.  
     [0687] In the q2d treatment schedule there was an increase in ALT levels over saline treated animals from 125 IU/L (saline control) to 300 IU/L (both PTEN oligonucleotide, ISIS 116847, and mismatch control), whereas AST levels remained between 220 IU/L and 240 IU/L among the three treatment groups.  
     [0688] In the q4d treatment schedule, ALT levels increased from 125 IU/L (saline control) to 160 IU/L in animals treated with ISIS 116847 and 200 IU/L for mismatch control. AST levels decreased from saline control levels of 220 IU/L to 160 IU/L for ISIS 116847 treated animals, as well as in animals treated with the mismatch control (200 IU/L). As a comparison, AST and ALT levels were measured after treatment with troglitazone. Levels of both enzymes were found to be 260 IU/L.  
     [0689] In a similar experiment, male db/db and wild-type mice were treated once a week for 4 weeks with ISIS 116847 at 10, 25, 50 or 100 mg/kg or ISIS 116845 at 50 mg/kg. Controls included saline or ISIS 29848 (the mixed sequence 20-mer negative oligonucleotide control, SEQ ID NO: 461) As a comparison db/db mice were also treated with troglitazone. At day 28 mice were sacrificed and AST and ALT levels were measured.  
     [0690] Treatment of db/db mice with ISIS 116847 resulted in a dose-dependent increase in ALT levels over the dose range with animals treated at the high dose having ALT levels of 250 IU/L while AST levels remained constant at 165 IU/L. These levels represent an increase in ALT levels from saline treated controls of 110 IU/L and a decrease in AST levels from saline treated controls of 220 IU/L. db/db animals treated with ISIS 116845 at a dose of 50 mg/kg had comparable ALT and AST levels, 145 IU/L. Animals treated with the universal control had ALT and AST levels comparable to control levels and those treated with troglitazone showed an increase in ALT levels over control to 150 IU/L and a slight decrease in AST levels to 200 IU/L from control.  
     [0691] Wild-type mice treated with 100 mg/kg of ISIS 116847 had both increased ALT and AST levels (100 IU/L and 130 IU/L, respectively) compared to saline-treated control ALT and AST levels (50 IU/L and 95 IU/L, respectively).  
     [0692] Although ALT levels were slightly elevated in animals treated with PTEN oligomeric compounds, AST levels were reduced indicating that PTEN oligomeric compound effects on liver weight were not due to toxicity.  
     Example 80  
     [0693] Design of Double Stranded Oligomeric Compounds Targeting PTEN  
     [0694] RNA interference (RNAi) and post-transcriptional gene silencing (PTGS) have become powerful and widely used tools for gene function analysis in invertebrates and plants (Fraser et al.,  Nature,  2000, 408, 325; Gönczy et al.  Nature,  2000, 408, 331). Introduction of double-stranded RNA (dsRNA) into the cells of these organisms leads to the sequence-specific degradation of homologous gene transcripts.  
     [0695] A number of PCT applications have recently been published that relate to the RNAi phenomenon. These include: PCT publication WO 00/44895; PCT publication WO 00/49035; PCT publication WO 00/63364; PCT publication WO 01/36641; PCT publication WO 01/36646; PCT publication WO 99/32619; PCT publication WO 00/44914; PCT publication WO 01/29058; and PCT publication WO 01/75164.  
     [0696] U.S. Pat. Nos. 5,898,031 and 6,107,094, each of which is commonly owned with this application and each of which is herein incorporated by reference, describe certain oligonucleotides having RNA-like properties. When hybridized with RNA, these oligonucleotides serve as substrates for a dsRNase enzyme with resultant cleavage of the target RNA by the enzyme.  
     [0697] In accordance with the present invention, a series of 21 nucleotide oligomeric compounds, in this case duplex RNAs (also known as small interfering RNAs: siRNAs), were designed to target PTEN mRNA (Genbank accession no. U92436. 1; SEQ ID NO: 828). The nucleobase sequence of the antisense strand of the duplex is identical to the 18 nucleobase oligonucleotides in Table 37 with one additional complementary base on the 3′ end of the oligoribonucleotides followed by a two-nucleobase overhang of deoxythymidine (T), TT. The sequences of the antisense strands are listed in Table 38. The sense strand of the dsRNAs listed in Table 39 were designed and synthesized as the complement of the antisense strands and also contained the two-nucleobase overhang on the 3′ end making both strands of the dsRNA duplex complementary over the central 19 nucleobases and each having a two-base overhang on the 3′ end. For example, the dsRNA having ISIS 29574 (SEQ ID NO: 832) as the antisense strand is:  
                                cgagaggcggacgggaccgTT   ISIS 29574             |||||||||||||||||||       TTgctctccgcctgccctggc   Complement of ISIS 29574          
 
     [0698] Both strands of the dsRNAs were purchased from Dharmacon Research Inc. (Lafayette, Colo.), shipped lyophilized and annealed on-site using the manufacturer&#39;s protocol. Briefly, each RNA oligonucleotide was aliquoted and diluted to a concentration of 50 μM. Once diluted, 30 uL of each strand was combined with 1.5 μL of a 5× solution of annealing buffer. The final concentration of said buffer was 100 mM potassium acetate, 30 mM HEPES-KOH pH 7.4, and 2mM magnesium acetate. The final volume was 75 μL. This solution was incubated for 1 minute at 90° C. and then centrifuged for 15 seconds. The tube was allowed to sit for 1 hour at 37° C. at which time the dsRNA duplexes were used in experimentation. The final concentration of the dsRNA duplex was 20 μM. This solution can be stored frozen (−20° C.) and freeze-thawed up to 5 times.  
     Example 81  
     [0699] Modulation of Human PTEN Expression by Double Stranded RNA (dsRNA)  
     [0700] In accordance with the present invention, a series of double stranded oligomeric compounds targeted to PTEN were evaluated for their ability to modulate PTEN expression in T-24 cells.  
     [0701] When cells reached 80% confluency, they were treated with dsRNA. For cells grown in 96-well plates, wells were washed once with 200 μL OPTI-MEM-1™ reduced-serum medium (Gibco BRL) and then treated with 130 μL of OPTI-MEM-1™ containing 12 μg/mL LIPOFECTIN™ (Gibco BRL) and the desired dsRNA at a final concentration of 200 nM. After 5 hours of treatment, the medium was replaced with fresh medium. Cells were harvested 16 hours after dsRNA or single-stranded oligonucleotide treatment, at which time RNA was isolated and target reduction measured by RT-PCR.  
     [0702] The sequences of the oligomeric compounds (antisense and sense to the PTEN target mRNA) of the dsRNAs are shown in Table 38 and 39, respectively. Prior to treatment of the T-24 cells, the dsRNA oligomers were generated by annealing the antisense and sense strands according to the method outlined in Example 80. Target sites are indicated by the first (5′ most) nucleotide number, as given in the sequence source reference (Genbank accession no. U92436.1), to which the antisense strand of the dsRNA oligonucleotide binds.  
     [0703] All compounds in Tables 38 and 39 are oligoribonucleotides, 21 nucleotides in length with the two nucleotides on the 3′ end being oligodeoxyribonucleotides, TT with phosphodiester backbones (internucleoside linkages) throughout. All oligoribonucleotides are depicted in the 5′→3′ direction.  
     [0704] Data were obtained by real-time quantitative PCR as described in other examples herein and are averaged from three experiments in which T-24 cells were cultured as described in Section 15 (15. Cell Lines for Assaying Oligonucleotide Activity) according treated with double stranded oligomeric compounds (composed of antisense strands SEQ ID NOs: 879-914 hybridized to sense strands: SEQ ID NOs: 915-950) targeting PTEN mRNA (SEQ ID NO: 828) according to the protocol outlined in Example 34. If present, “N.D.” indicates “no data”.  
                   TABLE 38                          Modulation of PTEN mRNA levels by dsRNA oligomers                                                 dsRNA                                   CORRESPONDING       TARGET       SEQUENCE OF ANTISENSE       TO ISIS#   REGION   SEQ ID NO   TARGET SITE   STRAND OF dsRNA   % INHIB   SEQ ID NO.                                                     29574   5′ UTR   828   19   cgagaggcggacgggaccgTT   0   876           29575   5′ UTR   828   57   cgggcgcctcggaagaccgTT   0   877       29576   5′ UTR   828   197   tggctgcagcttccgagagTT   40   878       29577   5′ UTR   828   314   cccgcggctgctcacaggcTT   25   879       29578   5′ UTR   828   421   caggagaagccgaggaagaTT   64   880       29579   5′ UTR   828   494   gggaggtgccgccgccgccTT   20   881       29581   5′ UTR   828   671   ccgggtccctggatgtgccTT   35   882       29582   5′ UTR   828   757   cctccgaacggctgcctccTT   59   883       29583   5′ UTR   828   817   tctcctcagcagccagaggTT   50   884       29584   5′ UTR   828   891   cgcttggctctggaccgcaTT   33   885       29585   5′ UTR   828   952   tcttctgcaggatggaaatTT   27   886       29587   Coding   828   1106   ggataaatataggtcaagtTT   49   887       29588   Coding   828   1169   tcaatattgttcctgtataTT   50   888       29589   Coding   828   1262   ttaaatttggcggtgtcatTT   64   889       29590   Coding   828   1342   caagatcttcacaaaagggTT   75   890       29591   Coding   828   1418   attacaccagttcgtccctTT   77   891       29592   Coding   828   1504   tgtctctggtccttacttcTT   76   892       29593   Coding   828   1541   acatagcgcctctgactggTT   74   893       29595   Coding   828   1694   gaatatatcttcacctttaTT   10   894       29596   Coding   828   1792   ggaagaactctactttgatTT   29   895       29597   Coding   828   1855   tgaagaatgtatttacccaTT   72   896       29599   Coding   828   2020   ggttggctttgtctttattTT   0   897       29600   Coding   828   2098   tgctagcctctggatttgaTT   43   898       29601   Coding   828   2180   tctggatcagagtcagtggTT   19   899       29602   3′ UTR   828   2268   tattttcatggtgttttacTT   59   900       29603   3′ UTR   828   2347   tgttcctataactggtaatTT   40   901       29604   3′ UTR   828   2403   gtgtcaaaaccctgtggatTT   45   902       29605   3′ UTR   828   2523   actggaataaaacgggaaaTT   38   903       29606   3′ UTR   828   2598   acttcagttggtgacagaaTT   25   904       29607   3′ UTR   828   2703   tagcaaaacctttcggaaaTT   31   905       29608   3′ UTR   828   2765   aattatttcctttctgagcTT   29   906       29609   3′ UTR   828   2806   taaatagctggagatggtcTT   7   907       29610   3′ UTR   828   2844   cagattaataactgtagcaTT   37   908       29611   3′ UTR   828   2950   ccccaatacagattcacttTT   39   909       29612   3′ UTR   828   3037   attgttgctgtgtttcttaTT   30   910       29613   3′ UTR   828   3088   tgtttcaagcccattctttTT   40   911                  
 
     [0705]                   TABLE 39                          Modulation of PTEN mRNA levels by dsRNA oligomers                                                 dsRNA                                   CORRESPONDING       TO COMPLEMENT       TARGET       SEQUENCE OF SENSE STRAND       OF ISIS#   REGION   SEQ ID NO   TARGET SITE   OF dsRNA   % INHIB   SEQ ID NO.                                                     29574   5′ UTR   828   19   cggtcccgtccgcctctcgTT   0   912           29575   5′ UTR   828   57   cggtcttccgaggcgcccgTT   0   913       29576   5′ UTR   828   197   ctctcggaagctgcagccaTT   40   914       29577   5′ UTR   828   314   gcctgtgagcagccgcgggTT   25   915       29578   5′ UTR   828   421   tcttcctcggcttctcctgTT   64   916       29579   5′ UTR   828   494   ggcggcggcggcacctcccTT   20   917       29581   5′ UTR   828   671   ggcacatccagggacccggTT   35   918       29582   5′ UTR   828   757   ggaggcagccgttcggaggTT   59   919       29583   5′ UTR   828   817   cctctggctgctgaggagaTT   50   920       29584   5′ UTR   828   891   tgcggtccagagccaagcgTT   33   921       29585   5′ UTR   828   952   atttccatcctgcagaagaTT   27   922       29587   Coding   828   1106   acttgacctatatttatccTT   49   923       29588   Coding   828   1169   tatacaggaacaatattgaTT   50   924       29589   Coding   828   1262   atgacaccgccaaatttaaTT   64   925       29590   Coding   828   1342   cccttttgtgaagatcttgTT   75   926       29591   Coding   828   1418   agggacgaactggtgtaatTT   77   927       29592   Coding   828   1504   gaagtaaggaccagagacaTT   76   928       29593   Coding   828   1541   ccagtcagaggcgctatgtTT   74   929       29595   Coding   828   1694   taaaggtgaagatatattcTT   10   930       29596   Coding   828   1792   atcaaagtagagttcttccTT   29   931       29597   Coding   828   1855   tgggtaaatacattcttcaTT   72   932       29599   Coding   828   2020   aataaagacaaagccaaccTT   0   933       29600   Coding   828   2098   tcaaatccagaggctagcaTT   43   934       29601   Coding   828   2180   ccactgactctgatccagaTT   19   935       29602   3′ UTR   828   2268   gtaaaacaccatgaaaataTT   59   936       29603   3′ UTR   828   2347   attaccagttataggaacaTT   40   937       29604   3′ UTR   828   2403   atccacagggttttgacacTT   45   938       29605   3′ UTR   828   2523   tttcccgttttattccagtTT   38   939       29606   3′ UTR   828   2598   ttctgtcaccaactgaagtTT   25   940       29607   3′ UTR   828   2703   tttccgaaaggttttgctaTT   31   941       29608   3′ UTR   828   2765   gctcagaaaggaaataattTT   29   942       29609   3′ UTR   828   2806   gaccatctccagctatttaTT   7   943       29610   3′ UTR   828   2844   tgctacagttattaatctgTT   37   944       29611   3′ UTR   828   2950   aagtgaatctgtattggggTT   39   945       29612   3′ UTR   828   3037   taagaaacacagcaacaatTT   30   946       29613   3′ UTR   828   3088   aaagaatgggcttgaaacaTT   40   947                    
     [0706] The antisense strands represented by SEQ ID NOs: 879, 880, 882, 883, 884, 885, 887, 888, 889, 890, 891, 892, 893, 896, 998, 900, 901, 902, 903, 905, 908, 909, 910 and 911 (Table 38) are from the preferred dsRNAs which demonstrated at least 30% inhibition of PTEN expression in this experiment. The corresponding sense strands of the preferred dsRNA oligomers are represented by SEQ ID NOs: 914, 916, 918, 919, 920, 921, 923, 924, 925, 926, 927, 928, 929, 932, 934, 936, 937, 938, 939, 941, 944, 945, 946 and 947 (Table 39).  
     [0707] The target sites to which these preferred sequences are complementary are herein referred to as, “preferred target segments” and are therefore preferred sites for targeting by compounds of the present invention.  
     [0708] One having skill in the art will recognize that the methods of identification of preferred dsRNA oligomeric compounds for modulation of PTEN expression outlined in this example may be applied to any target for the purpose of target validation or gene function analysis. It will also be recognizable to one skilled in the art, that for any particular target, screening of antisense oligonucleotides need not be carried out prior to design of and screening of dsRNA compounds. Thus, a plurality of virtual dsRNA compounds targeted to functional regions of any target can be generated and subjected to a selection process, actual compounds corresponding to a subset of virtual compounds may be robotically synthesized, and modulators can be identified which may subsequently employed in the processes of gene function analysis or target validation via the methods herein described.  
    
     
       
         1 
         
           
             947  
           
           
             1  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            1 

ccaggcggca ggaccact                                                   18 

 
           
             2  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            2 

gaccaggcgg caggacca                                                   18 

 
           
             3  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            3 

aggtgagacc aggcggca                                                   18 

 
           
             4  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            4 

cagaggcaga cgaaccat                                                   18 

 
           
             5  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            5 

gcagaggcag acgaacca                                                   18 

 
           
             6  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            6 

gcaagcagcc ccagagga                                                   18 

 
           
             7  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            7 

ggtcagcaag cagcccca                                                   18 

 
           
             8  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            8 

gacagcggtc agcaagca                                                   18 

 
           
             9  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            9 

gatggacagc ggtcagca                                                   18 

 
           
             10  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            10 

tctggatgga cagcggtc                                                   18 

 
           
             11  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            11 

ggtggttctg gatggaca                                                   18 

 
           
             12  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            12 

gtgggtggtt ctggatgg                                                   18 

 
           
             13  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            13 

gcagtgggtg gttctgga                                                   18 

 
           
             14  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            14 

cacaaagaac agcactga                                                   18 

 
           
             15  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            15 

ctggcacaaa gaacagca                                                   18 

 
           
             16  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            16 

tcctggctgg cacaaaga                                                   18 

 
           
             17  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            17 

ctgtcctggc tggcacaa                                                   18 

 
           
             18  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            18 

ctcaccagtt tctgtcct                                                   18 

 
           
             19  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            19 

tcactcacca gtttctgt                                                   18 

 
           
             20  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            20 

gtgcagtcac tcaccagt                                                   18 

 
           
             21  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            21 

actctgtgca gtcactca                                                   18 

 
           
             22  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            22 

cagtgaactc tgtgcagt                                                   18 

 
           
             23  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            23 

attccgtttc agtgaact                                                   18 

 
           
             24  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            24 

gaaggcattc cgtttcag                                                   18 

 
           
             25  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            25 

ttcaccgcaa ggaaggca                                                   18 

 
           
             26  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            26 

ctctgttcca ggtgtcta                                                   18 

 
           
             27  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            27 

ctggtggcag tgtgtctc                                                   18 

 
           
             28  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            28 

tggggtcgca gtatttgt                                                   18 

 
           
             29  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            29 

ggttggggtc gcagtatt                                                   18 

 
           
             30  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            30 

ctaggttggg gtcgcagt                                                   18 

 
           
             31  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            31 

ggtgcccttc tgctggac                                                   18 

 
           
             32  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            32 

ctgaggtgcc cttctgct                                                   18 

 
           
             33  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            33 

gtgtctgttt ctgaggtg                                                   18 

 
           
             34  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            34 

tggtgtctgt ttctgagg                                                   18 

 
           
             35  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            35 

acaggtgcag atggtgtc                                                   18 

 
           
             36  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            36 

ttcacaggtg cagatggt                                                   18 

 
           
             37  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            37 

gtgccagcct tcttcaca                                                   18 

 
           
             38  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            38 

tacagtgcca gccttctt                                                   18 

 
           
             39  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            39 

ggacacagct ctcacagg                                                   18 

 
           
             40  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            40 

tgcaggacac agctctca                                                   18 

 
           
             41  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            41 

gagcggtgca ggacacag                                                   18 

 
           
             42  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            42 

aagccgggcg agcatgag                                                   18 

 
           
             43  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            43 

aatctgcttg accccaaa                                                   18 

 
           
             44  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            44 

gaaacccctg tagcaatc                                                   18 

 
           
             45  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            45 

gtatcagaaa cccctgta                                                   18 

 
           
             46  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            46 

gctcgcagat ggtatcag                                                   18 

 
           
             47  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            47 

gcagggctcg cagatggt                                                   18 

 
           
             48  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            48 

tgggcagggc tcgcagat                                                   18 

 
           
             49  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            49 

gactgggcag ggctcgca                                                   18 

 
           
             50  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            50 

cattggagaa gaagccga                                                   18 

 
           
             51  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            51 

gatgacacat tggagaag                                                   18 

 
           
             52  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            52 

gcagatgaca cattggag                                                   18 

 
           
             53  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            53 

tcgaaagcag atgacaca                                                   18 

 
           
             54  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            54 

gtccaagggt gacatttt                                                   18 

 
           
             55  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            55 

cacagcttgt ccaagggt                                                   18 

 
           
             56  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            56 

ttggtctcac agcttgtc                                                   18 

 
           
             57  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            57 

caggtctttg gtctcaca                                                   18 

 
           
             58  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            58 

ctgttgcaca accaggtc                                                   18 

 
           
             59  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            59 

gtttgtgcct gcctgttg                                                   18 

 
           
             60  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            60 

gtcttgtttg tgcctgcc                                                   18 

 
           
             61  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            61 

ccacagacaa catcagtc                                                   18 

 
           
             62  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            62 

ctggggacca cagacaac                                                   18 

 
           
             63  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            63 

tcagccgatc ctggggac                                                   18 

 
           
             64  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            64 

caccaccagg gctctcag                                                   18 

 
           
             65  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            65 

gggatcacca ccagggct                                                   18 

 
           
             66  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            66 

gaggatggca aacaggat                                                   18 

 
           
             67  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            67 

accagcacca agaggatg                                                   18 

 
           
             68  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            68 

ttttgataaa gaccagca                                                   18 

 
           
             69  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            69 

tattggttgg cttcttgg                                                   18 

 
           
             70  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            70 

gggttcctgc ttggggtg                                                   18 

 
           
             71  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            71 

gtcgggaaaa ttgatctc                                                   18 

 
           
             72  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            72 

gatcgtcggg aaaattga                                                   18 

 
           
             73  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            73 

ggagccagga agatcgtc                                                   18 

 
           
             74  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            74 

tggagccagg aagatcgt                                                  18 

 
           
             75  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            75 

tggagcagca gtgttgga                                                   18 

 
           
             76  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            76 

gtaaagtctc ctgcactg                                                   18 

 
           
             77  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            77 

tggcatccat gtaaagtc                                                   18 

 
           
             78  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            78 

cggttggcat ccatgtaa                                                   18 

 
           
             79  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            79 

ctctttgcca tcctcctg                                                   18 

 
           
             80  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            80 

ctgtctctcc tgcactga                                                   18 

 
           
             81  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            81 

ggtgcagcct cactgtct                                                   18 

 
           
             82  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            82 

aactgcctgt ttgcccac                                                   18 

 
           
             83  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            83 

cttctgcctg cacccctg                                                   18 

 
           
             84  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            84 

actgactggg catagctc                                                   18 

 
           
             85  
             1240  
             DNA  
             Homo sapiens  
             
               CDS  
               (48)...(881)  
             
           
            85 

gcctcgctcg ggcgcccagt ggtcctgccg cctggtctca cctcgcc atg gtt cgt       56 
                                                    Met Val Arg 
                                                     1 

taa gct gcc tct gca gtg cgt cct ctg ggg ctg ctt gct gac cgc tgt      104 
 *  Ala Ala Ser Ala Val Arg Pro Leu Gly Leu Leu Ala Asp Arg Cys 
         5                   10                  15 

cca tuu gca utg cuu taa acc aga acc acc cac tgc atg cag aga aaa      152 
Pro Pro Ala Ala Leu  *  Thr Arg Thr Thr His Cys Met Gln Arg Lys 
     20                      25                  30 

aca gta cct aat aaa cag tgu taa cag gug tua cag tgc tgt tct ttg      200 
Thr Val Pro Asn Lys Gln Gln  *  Gln Val Val Gln Cys Cys Ser Leu 
     35                  40                      45 

tgc cag cca gga cag aaa ctg gtg agt gac tgc gcc ucg ggu aac aca      248 
Cys Gln Pro Gly Gln Lys Leu Val Ser Asp Cys Ala Ser Gly Asn Thr 
     50                  55                  60 

gag ttc act gaa acg gaa tgc ctt cct tgc ggt gaa agc gaa ttc tgu      296 
Glu Phe Thr Glu Thr Glu Cys Leu Pro Cys Gly Glu Ser Glu Phe Phe 
 65                  70                  75                  80 

tgu tgu cuc ggu guc tag aca cct gga aca gag aga cac act gcc acc      344 
Phe Phe Leu Gly Val  *  Thr Pro Gly Thr Glu Arg His Thr Ala Thr 
                 85                      90                  95 

agc aca aat act gcu att aag gut cgt cga ccc caa cct agg gct tcg      392 
Ser Thr Asn Thr Ala Ile Lys Lys Arg Arg Pro Gln Pro Arg Ala Ser 
                100                 105                 110 

ggt cca gca gaa ggg cac ctc aga aac aaa ugu aga ggg tgu tga cac      440 
Gly Pro Ala Glu Gly His Leu Arg Asn Lys Cys Arg Gly Gly  *  His 
            115                 120                 125 

cat ctg cac ctg tga aga agg ctg gca ctg tac gag tga ggc cat ctc      488 
His Leu His Leu  *  Arg Arg Leu Ala Leu Tyr Glu  *  Gly His Leu 
            130                     135                     140 

gug ugt ctg uaa tgt gag agc tgt gtc ctg cac cgc tca tgc tcg ccc      536 
Val Val Leu  *  Cys Glu Ser Cys Val Leu His Arg Ser Cys Ser Pro 
                    145                 150                 155 

ggc ttt ggg gtc cgu cau agc gga aag cag att gct aca ggg gtt tct      584 
Gly Phe Gly Val Arg His Ser Gly Lys Gln Ile Ala Thr Gly Val Ser 
                160                 165                 170 

gat acc atc tgc gag ccc tgc cca gaa tga atc guc gtc ggc ttc ttc      632 
Asp Thr Ile Cys Glu Pro Cys Pro Glu  *  Ile Val Val Gly Phe Phe 
            175                 180                     185 

tcc aat gtg tca tct gct ttc gaa aaa tgt cac cct aga aaa guc tgg      680 
Ser Asn Val Ser Ser Ala Phe Glu Lys Cys His Pro Arg Lys Val Trp 
            190                 195                 200 

aca agc tgt gag acc aaa gac ctg gtt gtg caa cag gca ggc aca ttc      728 
Thr Ser Cys Glu Thr Lys Asp Leu Val Val Gln Gln Ala Gly Thr Phe 
        205                 210                 215 

gut aua agg aag taa caa gac tga tgt tgt ctg tgg tcc cca gga tcg      776 
Phe Ile Arg Lys  *  Gln Asp  *  Cys Cys Leu Trp Ser Pro Gly Ser 
    220                         225                 230 

gct gag agc cct gat aaa cgg aag uag aau gtg gtg atc ccc atc atc      824 
Ala Glu Ser Pro Asp Lys Arg Lys  *  Asn Val Val Ile Pro Ile Ile 
        235                 240                     245 

ttc ggg atc ctg ttt gcc atc ctc ttg gtg aag uaa uua ctg gtc ttt      872 
Phe Gly Ile Leu Phe Ala Ile Leu Leu Val Lys  *  Leu Leu Val Phe 
        250                 255                     260 

atc aaa aag gtggccaaga agccaaccaa taaggccccc uaaaataaac              921 
Ile Lys Lys 
        265 

accccaagca ggaaccccag gagatcaatt ttcccgacga tcttcctggu gguaaauggc    981 

tccaacactg ctgctccagt gcaggagact ttacatggat gccaagataa aaaggutugc   1041 

gccggtcacc caggaggatg gcaaagagag tcgcatctca gtgcaggaga tgguagguag   1101 

agguagacag tgaggctgca cccacccagg agtgtggcca cgtgggcaaa cagagggcag   1161 

ttggccagag agcctggtgc tgctgctgca ggggtgcagg cagaagcggg gagctatgcc   1221 

cagtcagtgc cagcccctc                                                1240 

 
           
             86  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            86 

cagagttcac tgaaacggaa tgc                                             23 

 
           
             87  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            87 

ggtggcagtg tgtctctctg ttc                                             23 

 
           
             88  
             25  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            88 

ttccttgcgg tgaaagcgaa ttcct                                           25 

 
           
             89  
             19  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            89 

gaaggtgaag gtcggagtc                                                  19 

 
           
             90  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            90 

gaagatggtg atgggatttc                                                 20 

 
           
             91  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            91 

caagcttccc gttctcagcc                                                 20 

 
           
             92  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            92 

agtggtcctg ccgcctggtc                                                 20 

 
           
             93  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            93 

gaacagcact gactgttt                                                   18 

 
           
             94  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            94 

agaacagcac tgactgtt                                                   18 

 
           
             95  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            95 

aagaacagca ctgactgt                                                   18 

 
           
             96  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            96 

aaagaacagc actgactg                                                   18 

 
           
             97  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            97 

caaagaacag cactgact                                                   18 

 
           
             98  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            98 

acaaagaaca gcactgac                                                   18 

 
           
             99  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            99 

cacaaagaac agcactga                                                   18 

 
           
             100  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            100 

gcacaaagaa cagcactg                                                   18 

 
           
             101  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            101 

ggcacaaaga acagcact                                                   18 

 
           
             102  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            102 

tggcacaaag aacagcac                                                   18 

 
           
             103  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            103 

gctggcacaa agaacagc                                                   18 

 
           
             104  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            104 

ggctggcaca aagaacag                                                   18 

 
           
             105  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            105 

tggctggcac aaagaaca                                                   18 

 
           
             106  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            106 

ctggctggca caaagaac                                                   18 

 
           
             107  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            107 

cctggctggc acaaagaa                                                   18 

 
           
             108  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            108 

tcctggctgg cacaaaga                                                   18 

 
           
             109  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            109 

gtcctggctg gcacaaag                                                   18 

 
           
             110  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            110 

tgtcctggct ggcacaaa                                                   18 

 
           
             111  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            111 

ctgtcctggc tggcacaa                                                   18 

 
           
             112  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            112 

tctgtcctgg ctggcaca                                                   18 

 
           
             113  
             1058  
             DNA  
             Homo sapiens  
             
               CDS  
               (77)...(658)  
             
           
            113 

gccttgactt catctcagct ccagagcccg ccctctcttc ctgcagcctg ggaacttcag     60 

ccggctggag cccacc atg gct gca atc cga aag aag ctg gtg atc gtt ggg    112 
                  Met Ala Ala Ile Arg Lys Lys Leu Val Ile Val Gly 
                   1               5                   10 

gat ggt gcc tgt ggg aag acc tgc ctc ctc atc gtc ttc agc aag gat      160 
Asp Gly Ala Cys Gly Lys Thr Cys Leu Leu Ile Val Phe Ser Lys Asp 
         15                  20                  25 

cag ttt ccg gag gtc tac gtc cct act gtc ttt gag aac tat att gcg      208 
Gln Phe Pro Glu Val Tyr Val Pro Thr Val Phe Glu Asn Tyr Ile Ala 
     30                  35                  40 

gac att gag gtg gac ggc aag cag gtg gag ctg gct ctg tgg gac aca      256 
Asp Ile Glu Val Asp Gly Lys Gln Val Glu Leu Ala Leu Trp Asp Thr 
 45                  50                  55                  60 

gca ggg cag gaa gac tat gat cga ctg cgg cct ctc tcc tac ccg gac      304 
Ala Gly Gln Glu Asp Tyr Asp Arg Leu Arg Pro Leu Ser Tyr Pro Asp 
                 65                  70                  75 

act gat gtc atc ctc atg tgc ttc tcc atc gac agc cct gac agc ctg      352 
Thr Asp Val Ile Leu Met Cys Phe Ser Ile Asp Ser Pro Asp Ser Leu 
             80                  85                  90 

gaa aac att cct gag aag tgg acc cca gag gtg aag cac ttc tgc ccc      400 
Glu Asn Ile Pro Glu Lys Trp Thr Pro Glu Val Lys His Phe Cys Pro 
         95                 100                 105 

aac gtg ccc atc atc ctg gtg ggg aat aag aag gac ctg agg caa gac      448 
Asn Val Pro Ile Ile Leu Val Gly Asn Lys Lys Asp Leu Arg Gln Asp 
    110                 115                 120 

gag cac acc agg aga gag ctg gcc aag atg aag cag gag ccc gtt cgg      496 
Glu His Thr Arg Arg Glu Leu Ala Lys Met Lys Gln Glu Pro Val Arg 
125                 130                 135                 140 

tct gag gaa ggc cgg gac atg gcg aac cgg atc agt gcc ttt ggc tac      544 
Ser Glu Glu Gly Arg Asp Met Ala Asn Arg Ile Ser Ala Phe Gly Tyr 
                145                 150                 155 

ctt gag tgc tca gcc aag acc aag gag gga gtg cgg gag gtg ttt gag      592 
Leu Glu Cys Ser Ala Lys Thr Lys Glu Gly Val Arg Glu Val Phe Glu 
            160                 165                 170 

atg gcc act cgg gct ggc ctc cag gtc cgc aag aac aag cgt cgg agg      640 
Met Ala Thr Arg Ala Gly Leu Gln Val Arg Lys Asn Lys Arg Arg Arg 
        175                 180                 185 

ggc tgt ccc att ctc tga gatcccccca aagggccctt ttcctacatg             688 
Gly Cys Pro Ile Leu  * 
    190 

ccccctccct tcacaggggt acagaaatta tccccctaca accccagcct cctgagggct    748 

ccatactgaa ggctccattt tcagttccct cctgcccagg actgcattgt tttctagccc    808 

cgaggtgtgg cacgggccct ccctcccagc gctctgggag ccacgcctat gccctgccct    868 

tcctcatggg cccctgggga tcttgcccct ttgaccttcc ccaaaggatg gtcacacacc    928 

agcactttat acacttctgg ctcacaggaa agtgtctgca gtagggaccc agagtcccag    988 

gcccctggag ttgtttctgc aggggccttg tctctcactg catttggtca ggggggcatg   1048 

aataaaggct                                                          1058 

 
           
             114  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            114 

tgatgtcatc ctcatgtgct tct                                             23 

 
           
             115  
             19  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            115 

ccaggatgat gggcacgtt                                                  19 

 
           
             116  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            116 

cgacagccct gacagcctgg aaa                                             23 

 
           
             117  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            117 

gagctgagat gaagtcaa                                                   18 

 
           
             118  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            118 

gctgaagttc ccaggctg                                                   18 

 
           
             119  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            119 

ccggctgaag ttcccagg                                                   18 

 
           
             120  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            120 

ggcaccatcc ccaacgat                                                   18 

 
           
             121  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            121 

aggcaccatc cccaacga                                                   18 

 
           
             122  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            122 

tcccacaggc accatccc                                                   18 

 
           
             123  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            123 

aggtcttccc acaggcac                                                   18 

 
           
             124  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            124 

atgaggaggc aggtcttc                                                   18 

 
           
             125  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            125 

ttgctgaaga cgatgagg                                                   18 

 
           
             126  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            126 

tcaaagacag tagggacg                                                   18 

 
           
             127  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            127 

ttctcaaaga cagtaggg                                                   18 

 
           
             128  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            128 

agttctcaaa gacagtag                                                   18 

 
           
             129  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            129 

tgttttccag gctgtcag                                                   18 

 
           
             130  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            130 

tcgtcttgcc tcaggtcc                                                   18 

 
           
             131  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            131 

gtgtgctcgt cttgcctc                                                   18 

 
           
             132  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            132 

ctcctggtgt gctcgtct                                                   18 

 
           
             133  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            133 

cagaccgaac gggctcct                                                   18 

 
           
             134  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            134 

ttcctcagac cgaacggg                                                   18 

 
           
             135  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            135 

actcaaggta gccaaagg                                                   18 

 
           
             136  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            136 

ctcccgcact ccctcctt                                                   18 

 
           
             137  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            137 

ctcaaacacc tcccgcac                                                   18 

 
           
             138  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            138 

ggccatctca aacacctc                                                   18 

 
           
             139  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            139 

cttgttcttg cggacctg                                                   18 

 
           
             140  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            140 

cccctccgac gcttgttc                                                   18 

 
           
             141  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            141 

gtatggagcc ctcaggag                                                   18 

 
           
             142  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            142 

gagccttcag tatggagc                                                   18 

 
           
             143  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            143 

gaaaatggag ccttcagt                                                   18 

 
           
             144  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            144 

ggaactgaaa atggagcc                                                   18 

 
           
             145  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            145 

ggagggaact gaaaatgg                                                   18 

 
           
             146  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            146 

gcaggaggga actgaaaa                                                   18 

 
           
             147  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            147 

agggcagggc ataggcgt                                                   18 

 
           
             148  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            148 

ggaagggcag ggcatagg                                                   18 

 
           
             149  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            149 

catgaggaag ggcagggc                                                   18 

 
           
             150  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            150 

taaagtgctg gtgtgtga                                                   18 

 
           
             151  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            151 

cctgtgagcc agaagtgt                                                   18 

 
           
             152  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            152 

ttcctgtgag ccagaagt                                                   18 

 
           
             153  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            153 

cactttcctg tgagccag                                                   18 

 
           
             154  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            154 

agacactttc ctgtgagc                                                   18 

 
           
             155  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            155 

actctgggtc cctactgc                                                   18 

 
           
             156  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            156 

tgcagaaaca actccagg                                                   18 

 
           
             157  
             3076  
             DNA  
             Homo sapiens  
             
               CDS  
               (725)...(2539)  
             
           
            157 

gaattcaaaa tgtcttcagt tgtaaatctt accattattt tacgtacctc taagaaataa     60 

aagtgcttct aattaaaata tgatgtcatt aattatgaaa tacttcttga taacagaagt    120 

tttaaaatag ccatcttaga atcagtgaaa tatggtaatg tattattttc ctcctttgag    180 

ttaggtcttg tgcttttttt tcctggccac taaatttcac aatttccaaa aagcaaaata    240 

aacatattct gaatattttt gctgtgaaac acttgacagc agagctttcc accatgaaaa    300 

gaagcttcat gagtcacaca ttacatcttt gggttgattg aatgccactg aaacattcta    360 

gtagcctgga gaagttgacc tacctgtgga gatgcctgcc attaaatggc atcctgatgg    420 

cttaatacac atcactcttc tgtgaagggt tttaattttc aacacagctt actctgtagc    480 

atcatgttta cattgtatgt ataaagatta tacaaaggtg caattgtgta tttcttcctt    540 

aaaatgtatc agtataggat ttagaatctc catgttgaaa ctctaaatgc atagaaataa    600 

aaataataaa aaatttttca ttttggcttt tcagcctagt attaaaactg ataaaagcaa    660 

agccatgcac aaaactacct ccctagagaa aggctagtcc cttttcttcc ccattcattt    720 

catt atg aac ata gta gaa aac agc ata ttc tta tca aat ttg atg aaa     769 
     Met Asn Ile Val Glu Asn Ser Ile Phe Leu Ser Asn Leu Met Lys 
      1               5                   10                  15 

agc gcc aac acg ttt gaa ctg aaa tac gac ttg tca tgt gaa ctg tac      817 
Ser Ala Asn Thr Phe Glu Leu Lys Tyr Asp Leu Ser Cys Glu Leu Tyr 
                 20                  25                  30 

cga atg tct acg tat tcc act ttt cct gct ggg gtt cct gtc tca gaa      865 
Arg Met Ser Thr Tyr Ser Thr Phe Pro Ala Gly Val Pro Val Ser Glu 
             35                  40                  45 

agg agt ctt gct cgt gct ggt ttc tat tac act ggt gtg aat gac aag      913 
Arg Ser Leu Ala Arg Ala Gly Phe Tyr Tyr Thr Gly Val Asn Asp Lys 
         50                  55                  60 

gtc aaa tgc ttc tgt tgt ggc ctg atg ctg gat aac tgg aaa aga gga   961 
Val Lys Cys Phe Cys Cys Gly Leu Met Leu Asp Asn Trp Lys Arg Gly 
     65                  70                  75 

gac agt cct act gaa aag cat aaa aag ttg tat cct agc tgc aga ttc     1009 
Asp Ser Pro Thr Glu Lys His Lys Lys Leu Tyr Pro Ser Cys Arg Phe 
 80                  85                  90                  95 

gtt cag agt cta aat tcc gtt aac aac ttg gaa gct acc tct cag cct     1057 
Val Gln Ser Leu Asn Ser Val Asn Asn Leu Glu Ala Thr Ser Gln Pro 
                100                 105                 110 

act ttt cct tct tca gta aca aat tcc aca cac tca tta ctt ccg ggt     1105 
Thr Phe Pro Ser Ser Val Thr Asn Ser Thr His Ser Leu Leu Pro Gly 
            115                 120                 125 

aca gaa aac agt gga tat ttc cgt ggc tct tat tca aac tct cca tca     1153 
Thr Glu Asn Ser Gly Tyr Phe Arg Gly Ser Tyr Ser Asn Ser Pro Ser 
        130                 135                 140 

aat cct gta aac tcc aga gca aat caa gat ttt tct gcc ttg atg aga     1201 
Asn Pro Val Asn Ser Arg Ala Asn Gln Asp Phe Ser Ala Leu Met Arg 
    145                 150                 155 

agt tcc tac cac tgt gca atg aat aac gaa aat gcc aga tta ctt act     1249 
Ser Ser Tyr His Cys Ala Met Asn Asn Glu Asn Ala Arg Leu Leu Thr 
160                 165                 170                 175 

ttt cag aca tgg cca ttg act ttt ctg tcg cca aca gat ctg gca aaa     1297 
Phe Gln Thr Trp Pro Leu Thr Phe Leu Ser Pro Thr Asp Leu Ala Lys 
                180                 185                 190 

gca ggc ttt tac tac ata gga cct gga gac aga gtg gct tgc ttt gcc     1345 
Ala Gly Phe Tyr Tyr Ile Gly Pro Gly Asp Arg Val Ala Cys Phe Ala 
            195                 200                 205 

tgt ggt gga aaa ttg agc aat tgg gaa ccg aag gat aat gct atg tca     1393 
Cys Gly Gly Lys Leu Ser Asn Trp Glu Pro Lys Asp Asn Ala Met Ser 
        210                 215                 220 

gaa cac ctg aga cat ttt ccc aaa tgc cca ttt ata gaa aat cag ctt     1441 
Glu His Leu Arg His Phe Pro Lys Cys Pro Phe Ile Glu Asn Gln Leu 
    225                 230                 235 

caa gac act tca aga tac aca gtt tct aat ctg agc atg cag aca cat     1489 
Gln Asp Thr Ser Arg Tyr Thr Val Ser Asn Leu Ser Met Gln Thr His 
240                 245                 250                 255 

gca gcc cgc ttt aaa aca ttc ttt aac tgg ccc tct agt gtt cta gtt     1537 
Ala Ala Arg Phe Lys Thr Phe Phe Asn Trp Pro Ser Ser Val Leu Val 
                260                 265                 270 

aat cct gag cag ctt gca agt gcg ggt ttt tat tat gtg ggt aac agt     1585 
Asn Pro Glu Gln Leu Ala Ser Ala Gly Phe Tyr Tyr Val Gly Asn Ser 
            275                 280                 285 

gat gat gtc aaa tgc ttt tgc tgt gat ggt gga ctc agg tgt tgg gaa     1633 
Asp Asp Val Lys Cys Phe Cys Cys Asp Gly Gly Leu Arg Cys Trp Glu 
        290                 295                 300 

tct gga gat gat cca tgg gtt caa cat gcc aag tgg ttt cca agg tgt     1681 
Ser Gly Asp Asp Pro Trp Val Gln His Ala Lys Trp Phe Pro Arg Cys 
    305                 310                 315 

gag tac ttg ata aga att aaa gga cag gag ttc atc cgt caa gtt caa     1729 
Glu Tyr Leu Ile Arg Ile Lys Gly Gln Glu Phe Ile Arg Gln Val Gln 
320                 325                 330                 335 

gcc agt tac cct cat cta ctt gaa cag ctg cta tcc aca tca gac agc     1777 
Ala Ser Tyr Pro His Leu Leu Glu Gln Leu Leu Ser Thr Ser Asp Ser 
                340                 345                 350 

cca gga gat gaa aat gca gag tca tca att atc cat ttt gaa cct gga     1825 
Pro Gly Asp Glu Asn Ala Glu Ser Ser Ile Ile His Phe Glu Pro Gly 
            355                 360                 365 

gaa gac cat tca gaa gat gca atc atg atg aat act cct gtg att aat     1873 
Glu Asp His Ser Glu Asp Ala Ile Met Met Asn Thr Pro Val Ile Asn 
        370                 375                 380 

gct gcc gtg gaa atg ggc ttt agt aga agc ctg gta aaa cag aca gtt     1921 
Ala Ala Val Glu Met Gly Phe Ser Arg Ser Leu Val Lys Gln Thr Val 
    385                 390                 395 

caa aga aaa atc cta gca act gga gag aat tat aga cta gtc aat gat     1969 
Gln Arg Lys Ile Leu Ala Thr Gly Glu Asn Tyr Arg Leu Val Asn Asp 
400                 405                 410                 415 

ctt gtg tta gac tta ctc aat gca gaa gat gaa ata agg gaa gag gag     2017 
Leu Val Leu Asp Leu Leu Asn Ala Glu Asp Glu Ile Arg Glu Glu Glu 
                420                 425                 430 

aga gaa aga gca act gag gaa aaa gaa tca aat gat tta tta tta atc     2065 
Arg Glu Arg Ala Thr Glu Glu Lys Glu Ser Asn Asp Leu Leu Leu Ile 
            435                 440                 445 

cgg aag aat aga atg gca ctt ttt caa cat ttg act tgt gta att cca     2113 
Arg Lys Asn Arg Met Ala Leu Phe Gln His Leu Thr Cys Val Ile Pro 
        450                 455                 460 

atc ctg gat agt cta cta act gcc gga att att aat gaa caa gaa cat     2161 
Ile Leu Asp Ser Leu Leu Thr Ala Gly Ile Ile Asn Glu Gln Glu His 
    465                 470                 475 

gat gtt att aaa cag aag aca cag acg tct tta caa gca aga gaa ctg     2209 
Asp Val Ile Lys Gln Lys Thr Gln Thr Ser Leu Gln Ala Arg Glu Leu 
480                 485                 490                 495 

att gat acg att tta gta aaa gga aat att gca gcc act gta ttc aga     2257 
Ile Asp Thr Ile Leu Val Lys Gly Asn Ile Ala Ala Thr Val Phe Arg 
                500                 505                 510 

aac tct ctg caa gaa gct gaa gct gtg tta tat gag cat tta ttt gtg     2305 
Asn Ser Leu Gln Glu Ala Glu Ala Val Leu Tyr Glu His Leu Phe Val 
            515                 520                 525 

caa cag gac ata aaa tat att ccc aca gaa gat gtt tca gat cta cca     2353 
Gln Gln Asp Ile Lys Tyr Ile Pro Thr Glu Asp Val Ser Asp Leu Pro 
        530                 535                 540 

gtg gaa gaa caa ttg cgg aga cta caa gaa gaa aga aca tgt aaa gtg     2401 
Val Glu Glu Gln Leu Arg Arg Leu Gln Glu Glu Arg Thr Cys Lys Val 
    545                 550                 555 

tgt atg gac aaa gaa gtg tcc ata gtg ttt att cct tgt ggt cat cta     2449 
Cys Met Asp Lys Glu Val Ser Ile Val Phe Ile Pro Cys Gly His Leu 
560                 565                 570                 575 

gta gta tgc aaa gat tgt gct cct tct tta aga aag tgt cct att tgt     2497 
Val Val Cys Lys Asp Cys Ala Pro Ser Leu Arg Lys Cys Pro Ile Cys 
                580                 585                 590 

agg agt aca atc aag ggt aca gtt cgt aca ttt ctt tca tga             2539 
Arg Ser Thr Ile Lys Gly Thr Val Arg Thr Phe Leu Ser  * 
            595                 600 

agaagaacca aaacatcatc taaactttag aattaattta ttaaatgtat tataacttta   2599 

acttttatcc taatttggtt tccttaaaat ttttatttat ttacaactca aaaaacattg   2659 

ttttgtgtaa catatttata tatgtatcta aaccatatga acatatattt tttagaaact   2719 

aagagaatga taggcttttg ttcttatgaa cgaaaaagag gtagcactac aaacacaata   2779 

ttcaatcaaa atttcagcat tattgaaatt gtaagtgaag taaaacttaa gatatttgag   2839 

ttaaccttta agaattttaa atattttggc attgtactaa tacctggttt tttttttgtt   2899 

ttgttttttt gtacagacag ggcagcatac tgagaccctg cctttaaaaa caaacagaac   2959 

aaaaacaaaa caccagggac acatttctct gtcttttttg atcagtgtcc tatacatcga   3019 

aggtgtgcat atatgttgaa tgacatttta gggacatggt gtttttataa agaattc      3076 

 
           
             158  
             22  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            158 

ggactcaggt gttgggaatc tg                                              22 

 
           
             159  
             24  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            159 

caagtactca caccttggaa acca                                            24 

 
           
             160  
             27  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            160 

agatgatcca tgggttcaac atgccaa                                         27 

 
           
             161  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            161 

actgaagaca ttttgaat                                                   18 

 
           
             162  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            162 

cttagaggta cgtaaaat                                                   18 

 
           
             163  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            163 

gcacttttat ttcttaga                                                   18 

 
           
             164  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            164 

attttaatta gaagcact                                                   18 

 
           
             165  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            165 

accatatttc actgattc                                                   18 

 
           
             166  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            166 

ctaactcaaa ggaggaaa                                                   18 

 
           
             167  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            167 

cacaagacct aactcaaa                                                   18 

 
           
             168  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            168 

gctctgctgt caagtgtt                                                   18 

 
           
             169  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            169 

tgtgtgactc atgaagct                                                   18 

 
           
             170  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            170 

ttcagtggca ttcaatca                                                   18 

 
           
             171  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            171 

cttctccagg ctactaga                                                   18 

 
           
             172  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            172 

ggtcaacttc tccaggct                                                   18 

 
           
             173  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            173 

taaaaccctt cacagaag                                                   18 

 
           
             174  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            174 

ttaaggaaga aatacaca                                                   18 

 
           
             175  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            175 

gcatggcttt gcttttat                                                   18 

 
           
             176  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            176 

caaacgtgtt ggcgcttt                                                   18 

 
           
             177  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            177 

agcaggaaaa gtggaata                                                   18 

 
           
             178  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            178 

ttaacggaat ttagactc                                                   18 

 
           
             179  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            179 

atttgttact gaagaagg                                                   18 

 
           
             180  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            180 

agagccacgg aaatatcc                                                   18 

 
           
             181  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            181 

aaatcttgat ttgctctg                                                   18 

 
           
             182  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            182 

gtaagtaatc tggcattt                                                   18 

 
           
             183  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            183 

agcaagccac tctgtctc                                                   18 

 
           
             184  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            184 

tgaagtgtct tgaagctg                                                   18 

 
           
             185  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            185 

tttgacatca tcactgtt                                                   18 

 
           
             186  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            186 

tggcttgaac ttgacgga                                                   18 

 
           
             187  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            187 

tcatctcctg ggctgtct                                                   18 

 
           
             188  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            188 

gcagcattaa tcacagga                                                   18 

 
           
             189  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            189 

tttctctctc ctcttccc                                                   18 

 
           
             190  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            190 

aacatcatgt tcttgttc                                                   18 

 
           
             191  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            191 

atataacaca gcttcagc                                                   18 

 
           
             192  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            192 

aattgttctt ccactggt                                                   18 

 
           
             193  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            193 

aagaaggagc acaatctt                                                   18 

 
           
             194  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            194 

gaaaccaaat taggataa                                                   18 

 
           
             195  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            195 

tgtagtgcta cctctttt                                                   18 

 
           
             196  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            196 

ctgaaatttt gattgaat                                                   18 

 
           
             197  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            197 

tacaatttca ataatgct                                                   18 

 
           
             198  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            198 

gggtctcagt atgctgcc                                                   18 

 
           
             199  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            199 

ccttcgatgt ataggaca                                                   18 

 
           
             200  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            200 

catgtcccta aaatgtca                                                   18 

 
           
             201  
             2266  
             DNA  
             Homo sapiens  
             
               CDS  
               (316)...(1602)  
             
           
            201 

aattccgagc tgtagggaaa cgcaggggcg gcttctaggt gctgccgccg ccaccgccac     60 

caccacctcc accgccgcct cggaacccag gcctgggggg cggtggggcc gcgtatggag    120 

cccccgcccc ccggagctgc caacattgcc aacgccaccg ccacgctaca cacagcctca    180 

actttcagga gacccgtccg tggccttatt tattccaccc ttcctgtaca tcgtagcgaa    240 

tcaatccgtg gcgccgcact cctccgcatc cctctttaac agtacccctg ggatggcgtg    300 

agcactcccc cagcg atg gac cca tct gtg acg ctg tgg cag ttt ctg ctg     351 
                 Met Asp Pro Ser Val Thr Leu Trp Gln Phe Leu Leu 
                  1               5                   10 

cag ctg ctg aga gag caa ggc aat ggc cac atc atc tcc tgg act tca      399 
Gln Leu Leu Arg Glu Gln Gly Asn Gly His Ile Ile Ser Trp Thr Ser 
         15                  20                  25 

cgg gat ggt ggt gaa ttc aag ctg gtg gat gca gag gag gtg gcc cgg      447 
Arg Asp Gly Gly Glu Phe Lys Leu Val Asp Ala Glu Glu Val Ala Arg 
     30                  35                  40 

ctg tgg gga cta cgc aag aac aag acc aac atg aat tac gac aag ctc      495 
Leu Trp Gly Leu Arg Lys Asn Lys Thr Asn Met Asn Tyr Asp Lys Leu 
 45                  50                  55                  60 

agc cgg gcc ttg cgg tac tac tat gac aag aac atc atc cgc aag gtg      543 
Ser Arg Ala Leu Arg Tyr Tyr Tyr Asp Lys Asn Ile Ile Arg Lys Val 
                 65                  70                  75 

agc ggc cag aag ttc gtc tac aag ttt gtg tcc tac cct gag gtc gca      591 
Ser Gly Gln Lys Phe Val Tyr Lys Phe Val Ser Tyr Pro Glu Val Ala 
             80                  85                  90 

ggg tgc tcc act gag gac tgc ccg ccc cag cca gag gtg tct gtt acc      639 
Gly Cys Ser Thr Glu Asp Cys Pro Pro Gln Pro Glu Val Ser Val Thr 
         95                 100                 105 

tcc acc atg cca aat gtg gcc cct gct gct ata cat gcc gcc cca ggg      687 
Ser Thr Met Pro Asn Val Ala Pro Ala Ala Ile His Ala Ala Pro Gly 
    110                 115                 120 

gac act gtc tct gga aag cca ggc aca ccc aag ggt gca gga atg gca      735 
Asp Thr Val Ser Gly Lys Pro Gly Thr Pro Lys Gly Ala Gly Met Ala 
125                 130                 135                 140 

ggc cca ggc ggt ttg gca cgc agc agc cgg aac gag tac atg cgc tcg      783 
Gly Pro Gly Gly Leu Ala Arg Ser Ser Arg Asn Glu Tyr Met Arg Ser 
                145                 150                 155 

ggc ctc tat tcc acc ttc acc atc cag tct ctg cag ccg cag cca ccc      831 
Gly Leu Tyr Ser Thr Phe Thr Ile Gln Ser Leu Gln Pro Gln Pro Pro 
            160                 165                 170 

cct cat cct cgg cct gct gtg gtg ctc ccc aat gca gct cct gca ggg      879 
Pro His Pro Arg Pro Ala Val Val Leu Pro Asn Ala Ala Pro Ala Gly 
        175                 180                 185 

gca gca gcg ccc ccc tcg ggg agc agg agc acc agt cca agc ccc ttg      927 
Ala Ala Ala Pro Pro Ser Gly Ser Arg Ser Thr Ser Pro Ser Pro Leu 
    190                 195                 200 

gag gcc tgt ctg gag gct gaa gag gcc ggc ttg cct ctg cag gtc atc      975 
Glu Ala Cys Leu Glu Ala Glu Glu Ala Gly Leu Pro Leu Gln Val Ile 
205                 210                 215                 220 

ctg acc ccg ccc gag gcc cca aac ctg aaa tcg gaa gag ctt aat gtg     1023 
Leu Thr Pro Pro Glu Ala Pro Asn Leu Lys Ser Glu Glu Leu Asn Val 
                225                 230                 235 

gag ccg ggt ttg ggc cgg gct ttg ccc cca gaa gtg aaa gta gaa ggg     1071 
Glu Pro Gly Leu Gly Arg Ala Leu Pro Pro Glu Val Lys Val Glu Gly 
            240                 245                 250 

ccc aag gaa gag ttg gaa gtt gcg ggg gag aga ggg ttt gtg cca gaa     1119 
Pro Lys Glu Glu Leu Glu Val Ala Gly Glu Arg Gly Phe Val Pro Glu 
        255                 260                 265 

acc acc aag gcc gag cca gaa gtc cct cca cag gag ggc gtg cca gcc     1167 
Thr Thr Lys Ala Glu Pro Glu Val Pro Pro Gln Glu Gly Val Pro Ala 
    270                 275                 280 

cgg ctg ccc gcg gtt gtt atg gac acc gca ggg cag gcg ggc ggc cat     1215 
Arg Leu Pro Ala Val Val Met Asp Thr Ala Gly Gln Ala Gly Gly His 
285                 290                 295                 300 

gcg gct tcc agc cct gag atc tcc cag ccg cag aag ggc cgg aag ccc     1263 
Ala Ala Ser Ser Pro Glu Ile Ser Gln Pro Gln Lys Gly Arg Lys Pro 
                305                 310                 315 

cgg gac cta gag ctt cca ctc agc ccg agc ctg cta ggt ggg ccg gga     1311 
Arg Asp Leu Glu Leu Pro Leu Ser Pro Ser Leu Leu Gly Gly Pro Gly 
            320                 325                 330 

ccc gaa cgg acc cca gga tcg gga agt ggc tcc ggc ctc cag gct ccg     1359 
Pro Glu Arg Thr Pro Gly Ser Gly Ser Gly Ser Gly Leu Gln Ala Pro 
        335                 340                 345 

ggg ccg gcg ctg acc cca tcc ctg ctt cct acg cat aca ttg acc ccg     1407 
Gly Pro Ala Leu Thr Pro Ser Leu Leu Pro Thr His Thr Leu Thr Pro 
    350                 355                 360 

gtg ctg ctg aca ccc agc tcg ctg cct cct agc att cac ttc tgg agc     1455 
Val Leu Leu Thr Pro Ser Ser Leu Pro Pro Ser Ile His Phe Trp Ser 
365                 370                 375                 380 

acc ctg agt ccc att gcg ccc cgt agc ccg gcc aag ctc tcc ttc cag     1503 
Thr Leu Ser Pro Ile Ala Pro Arg Ser Pro Ala Lys Leu Ser Phe Gln 
                385                 390                 395 

ttt cca tcc agt ggc agc gcc cag gtg cac atc cct tct atc agc gtg     1551 
Phe Pro Ser Ser Gly Ser Ala Gln Val His Ile Pro Ser Ile Ser Val 
            400                 405                 410 

gat ggc ctc tcg acc ccc gtg gtg ctc tcc cca ggg ccc cag aag cca     1599 
Asp Gly Leu Ser Thr Pro Val Val Leu Ser Pro Gly Pro Gln Lys Pro 
        415                 420                 425 

tga ctactaccac caccaccacc accccttctg gggtcactcc atccatgctc          1652 

tctccagcca gccatctcaa ggagaaacat agttcaactg aaagactcat gctctgattg   1712 

tggtggggtg gggatccttg ggaagaatta ctcccaagag taactctcat tatctcctcc   1772 

acagaaaaca cacagcttcc acaacttctc tgttttctgt cagtccccca gtggccgccc   1832 

ttacacgtct cctacttcaa tggtaggggc ggtttattta tttatttttt gaaggccact   1892 

gggatgagcc tgacctaacc ttttagggtg gttaggacat ctcccccacc tccccacttt   1952 

tttccccaag acaagacaat cgaggtctgg cttgagaacg acctttcttt ctttatttct   2012 

cagcctgccc ttggggagat gagggagccc tgtctgcgtt tttggatgtg agtagaagag   2072 

ttagtttgtt ttgttttatt attcctggcc atactcaggg gtccaggaag aatttgtacc   2132 

atttaatggg ttgggagtct tggccaagga agaatcacac ccttggaata gaaatttcca   2192 

cctcccccaa cctttctctc agacagctta tcctttttca accaactttt tggccaggga   2252 

ggaatgtccc tttt                                                     2266 

 
           
             202  
             18  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            202 

gcaaggcaat ggccacat                                                   18 

 
           
             203  
             21  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            203 

ctcctctgca tccaccagct t                                               21 

 
           
             204  
             26  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            204 

tctcctggac ttcacgggat ggtggt                                          26 

 
           
             205  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            205 

cccctgcgtt tccctaca                                                   18 

 
           
             206  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            206 

ggtggtggtg gcggtggc                                                   18 

 
           
             207  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            207 

ggcgttggca atgttggc                                                   18 

 
           
             208  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            208 

aagttgaggc tgtgtgta                                                   18 

 
           
             209  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            209 

aggccacgga cgggtctc                                                   18 

 
           
             210  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            210 

gattgattcg ctacgatg                                                   18 

 
           
             211  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            211 

gggatgcgga ggagtgcg                                                   18 

 
           
             212  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            212 

agtgctcacg ccatccca                                                   18 

 
           
             213  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            213 

aaactgccac agcgtcac                                                   18 

 
           
             214  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            214 

gaagtccagg agatgatg                                                   18 

 
           
             215  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            215 

caccaccatc ccgtgaag                                                   18 

 
           
             216  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            216 

tcttgttctt gcgtagtc                                                   18 

 
           
             217  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            217 

tgttcttgtc atagtagt                                                   18 

 
           
             218  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            218 

tcaccttgcg gatgatgt                                                   18 

 
           
             219  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            219 

gagcaccctg cgacctca                                                   18 

 
           
             220  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            220 

ggcgggcagt cctcagtg                                                   18 

 
           
             221  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            221 

ggtgaaggtg gaatagag                                                   18 

 
           
             222  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            222 

tccgatttca ggtttggg                                                   18 

 
           
             223  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            223 

ttggtggttt ctggcaca                                                   18 

 
           
             224  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            224 

tggagggact tctggctc                                                   18 

 
           
             225  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            225 

gcgtaggaag cagggatg                                                   18 

 
           
             226  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            226 

gtgctccaga agtgaatg                                                   18 

 
           
             227  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            227 

actggatgga aactggaa                                                   18 

 
           
             228  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            228 

ggccatccac gctgatag                                                   18 

 
           
             229  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            229 

ccaccacaat cagagcat                                                   18 

 
           
             230  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            230 

gatccccacc ccaccaca                                                   18 

 
           
             231  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            231 

tgttttctgt ggaggaga                                                   18 

 
           
             232  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            232 

aaacagagaa gttgtgga                                                   18 

 
           
             233  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            233 

gggactgaca gaaaacag                                                   18 

 
           
             234  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            234 

ataaataaat aaaccgcc                                                   18 

 
           
             235  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            235 

gttaggtcag gctcatcc                                                   18 

 
           
             236  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            236 

gttctcaagc cagacctc                                                   18 

 
           
             237  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            237 

aataaagaaa gaaaggtc                                                   18 

 
           
             238  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            238 

agggcaggct gagaaata                                                   18 

 
           
             239  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            239 

cttctactca catccaaa                                                   18 

 
           
             240  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            240 

caaaacaaac taactctt                                                   18 

 
           
             241  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            241 

ggaataataa aacaaaac                                                   18 

 
           
             242  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            242 

ttcttcctgg acccctga                                                   18 

 
           
             243  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            243 

ccaagggtgt gattcttc                                                   18 

 
           
             244  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            244 

tgtctgagag aaaggttg                                                   18 

 
           
             245  
             1080  
             DNA  
             Homo sapiens  
             
               CDS  
               (1)...(1080)  
             
           
            245 

atg act ctg gag tcc atc atg gcg tgt tgc ctg agc gat gag gtg aag       48 
Met Thr Leu Glu Ser Ile Met Ala Cys Cys Leu Ser Asp Glu Val Lys 
 1               5                   10                  15 

gag tcc aag cgg atc aac gcc gag atc gag aag cag ctg cgg cgg gac       96 
Glu Ser Lys Arg Ile Asn Ala Glu Ile Glu Lys Gln Leu Arg Arg Asp 
             20                  25                  30 

aag cgc gac gcc cgg cgc gag ctc aag ctg ctg ctg ctc ggc acg ggc      144 
Lys Arg Asp Ala Arg Arg Glu Leu Lys Leu Leu Leu Leu Gly Thr Gly 
         35                  40                  45 

gag agc ggg aag agc acg ttc atc aag cag atg cgc atc atc cac ggc      192 
Glu Ser Gly Lys Ser Thr Phe Ile Lys Gln Met Arg Ile Ile His Gly 
     50                  55                  60 

gcc ggc tac tcg gag gag gac aag cgc ggc ttc acc aag ctc gtc tac      240 
Ala Gly Tyr Ser Glu Glu Asp Lys Arg Gly Phe Thr Lys Leu Val Tyr 
 65                  70                  75                  80 

cag aac atc ttc acc gcc atg cag gcc atg atc cgg gcc atg gag acg      288 
Gln Asn Ile Phe Thr Ala Met Gln Ala Met Ile Arg Ala Met Glu Thr 
                 85                  90                  95 

ctc aag atc ctc tac aag tac gag cag aac aag gcc aat gcg ctc ctg      336 
Leu Lys Ile Leu Tyr Lys Tyr Glu Gln Asn Lys Ala Asn Ala Leu Leu 
            100                 105                 110 

atc cgg gag gtg gac gtg gag aag gtg acc acc ttc gag cat cag tac      384 
Ile Arg Glu Val Asp Val Glu Lys Val Thr Thr Phe Glu His Gln Tyr 
        115                 120                 125 

gtc agt gcc atc aag acc ctg tgg gag gac ccg ggc atc cag gaa tgc      432 
Val Ser Ala Ile Lys Thr Leu Trp Glu Asp Pro Gly Ile Gln Glu Cys 
    130                 135                 140 

tac gac cgc agg cgc gag tac cag ctc tcc gac tct gcc aag tac tac      480 
Tyr Asp Arg Arg Arg Glu Tyr Gln Leu Ser Asp Ser Ala Lys Tyr Tyr 
145                 150                 155                 160 

ctg acc gac gtt gac cgc atc gcc acc ttg ggc tac ctg ccc acc cag      528 
Leu Thr Asp Val Asp Arg Ile Ala Thr Leu Gly Tyr Leu Pro Thr Gln 
                165                 170                 175 

cag gac gtg ctg cgg gtc cgc gtg ccc acc acc ggc atc atc gag tac      576 
Gln Asp Val Leu Arg Val Arg Val Pro Thr Thr Gly Ile Ile Glu Tyr 
            180                 185                 190 

cct ttc gac ctg gag aac atc atc ttc cgg atg gtg gat gtg ggg ggc      624 
Pro Phe Asp Leu Glu Asn Ile Ile Phe Arg Met Val Asp Val Gly Gly 
        195                 200                 205 

cag cgg tcg gag cgg agg aag tgg atc cac tgc ttt gag aac gtg aca      672 
Gln Arg Ser Glu Arg Arg Lys Trp Ile His Cys Phe Glu Asn Val Thr 
    210                 215                 220 

tcc atc atg ttt ctc gtc gcc ctc agc gaa tac gac caa gtc ctg gtg      720 
Ser Ile Met Phe Leu Val Ala Leu Ser Glu Tyr Asp Gln Val Leu Val 
225                 230                 235                 240 

gag tcg gac aac gag aac cgg atg gag gag agc aaa gcc ctg ttc cgg      768 
Glu Ser Asp Asn Glu Asn Arg Met Glu Glu Ser Lys Ala Leu Phe Arg 
                245                 250                 255 

acc atc atc acc tac ccc tgg ttc cag aac tcc tcc gtc atc ctc ttc      816 
Thr Ile Ile Thr Tyr Pro Trp Phe Gln Asn Ser Ser Val Ile Leu Phe 
            260                 265                 270 

ctc aac aag aag gac ctg ctg gag gac aag atc ctg tac tcg cac ctg      864 
Leu Asn Lys Lys Asp Leu Leu Glu Asp Lys Ile Leu Tyr Ser His Leu 
        275                 280                 285 

gtg gac tac ttc ccc gag ttc gat ggt ccc cag cgg gac gcc cag gcg      912 
Val Asp Tyr Phe Pro Glu Phe Asp Gly Pro Gln Arg Asp Ala Gln Ala 
    290                 295                 300 

gcg cgg gag ttc atc ccg aag atg ttc gtg gac ctg aac ccc gac agc      960 
Ala Arg Glu Phe Ile Pro Lys Met Phe Val Asp Leu Asn Pro Asp Ser 
305                 310                 315                 320 

gac aag atc atc tac tca cac ttc acg tgt gcc acc gac acg gag aac     1008 
Asp Lys Ile Ile Tyr Ser His Phe Thr Cys Ala Thr Asp Thr Glu Asn 
                325                 330                 335 

atc cgc ttc gtg ttc gcg gcc gtg aag gac acc atc ctg cag ctg aac     1056 
Ile Arg Phe Val Phe Ala Ala Val Lys Asp Thr Ile Leu Gln Leu Asn 
            340                 345                 350 

ctg aag gag tac aat ctg gtc taa                                     1080 
Leu Lys Glu Tyr Asn Leu Val  * 
        355  
           
             246  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            246 

tgaccacctt cgagcatcag                                                 20 

 
           
             247  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            247 

cggtcgtagc attcctggat                                                 20 

 
           
             248  
             26  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            248 

tcagtgccat caagaccctg tgggag                                          26 

 
           
             249  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            249 

gatggactcc agagtcat                                                   18 

 
           
             250  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            250 

gccatgatgg actccaga                                                   18 

 
           
             251  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            251 

cacgccatga tggactcc                                                   18 

 
           
             252  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            252 

ctcatcgctc aggcaaca                                                   18 

 
           
             253  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            253 

cttcacctca tcgctcag                                                   18 

 
           
             254  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            254 

gactccttca cctcatcg                                                   18 

 
           
             255  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            255 

atccgcttgg actccttc                                                   18 

 
           
             256  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            256 

cgttgatccg cttggact                                                   18 

 
           
             257  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            257 

ctcgatctcg gcgttgat                                                   18 

 
           
             258  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            258 

cccgccgcag ctgcttct                                                   18 

 
           
             259  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            259 

cttgagctcg cgccgggc                                                   18 

 
           
             260  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            260 

gcagcagcag cttgagct                                                   18 

 
           
             261  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            261 

gcccgtgccg agcagcag                                                   18 

 
           
             262  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            262 

acgtgctctt cccgctct                                                   18 

 
           
             263  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            263 

atctgcttga tgaacgtg                                                   18 

 
           
             264  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            264 

cgcatctgct tgatgaac                                                   18 

 
           
             265  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            265 

gtagccggcg ccgtggat                                                   18 

 
           
             266  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            266 

tgtcctcctc cgagtagc                                                   18 

 
           
             267  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            267 

cttgtcctcc tccgagta                                                   18 

 
           
             268  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            268 

aagccgcgct tgtcctcc                                                   18 

 
           
             269  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            269 

tagacgagct tggtgaag                                                   18 

 
           
             270  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            270 

tgttctggta gacgagct                                                   18 

 
           
             271  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            271 

tggcggtgaa gatgttct                                                   18 

 
           
             272  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            272 

cggatcatgg cctgcatg                                                   18 

 
           
             273  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            273 

cgtctccatg gcccggat                                                   18 

 
           
             274  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            274 

tagaggatct tgagcgtc                                                   18 

 
           
             275  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            275 

tgtagaggat cttgagcg                                                   18 

 
           
             276  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            276 

tgctcgtact tgtagagg                                                   18 

 
           
             277  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            277 

gccttgttct gctcgtac                                                   18 

 
           
             278  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            278 

ttggccttgt tctgctcg                                                   18 

 
           
             279  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            279 

caggagcgca ttggcctt                                                   18 

 
           
             280  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            280 

ctccacgtcc acctcccg                                                   18 

 
           
             281  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            281 

ggtcaccttc tccacgtc                                                   18 

 
           
             282  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            282 

gatgctcgaa ggtggtca                                                   18 

 
           
             283  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            283 

actgacgtac tgatgctc                                                   18 

 
           
             284  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            284 

cttgatggca ctgacgta                                                   18 

 
           
             285  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            285 

cagggtcttg atggcact                                                   18 

 
           
             286  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            286 

ctggatgccc gggtcctc                                                   18 

 
           
             287  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            287 

tcctggatgc ccgggtcc                                                   18 

 
           
             288  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            288 

cgcctgcggt cgtagcat                                                   18 

 
           
             289  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            289 

gctggtactc gcgcctgc                                                   18 

 
           
             290  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            290 

tacttggcag agtcggag                                                   18 

 
           
             291  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            291 

gtcaggtagt acttggca                                                   18 

 
           
             292  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            292 

ggtcaacgtc ggtcaggt                                                   18 

 
           
             293  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            293 

gtggcgatgc ggtcaacg                                                   18 

 
           
             294  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            294 

gcaggtagcc caaggtgg                                                   18 

 
           
             295  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            295 

cgtcctgctg ggtgggca                                                   18 

 
           
             296  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            296 

ggtggtgggc acgcggac                                                   18 

 
           
             297  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            297 

tcgatgatgc cggtggtg                                                   18 

 
           
             298  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            298 

ccaggtcgaa agggtact                                                   18 

 
           
             299  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            299 

tgttctccag gtcgaaag                                                   18 

 
           
             300  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            300 

agatgatgtt ctccaggt                                                   18 

 
           
             301  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            301 

atccggaaga tgatgttc                                                   18 

 
           
             302  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            302 

ctccgctccg accgctgg                                                   18 

 
           
             303  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            303 

gatccacttc ctccgctc                                                   18 

 
           
             304  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            304 

tgtcacgttc tcaaagca                                                   18 

 
           
             305  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            305 

atgatggatg tcacgttc                                                   18 

 
           
             306  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            306 

cgagaaacat gatggatg                                                   18 

 
           
             307  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            307 

gctgagggcg acgagaaa                                                   18 

 
           
             308  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            308 

cgactccacc aggacttg                                                   18 

 
           
             309  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            309 

atccggttct cgttgtcc                                                   18 

 
           
             310  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            310 

ccatccggtt ctcgttgt                                                   18 

 
           
             311  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            311 

agggctttgc tctcctcc                                                   18 

 
           
             312  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            312 

ggtccggaac agggcttt                                                   18 

 
           
             313  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            313 

gtaggtgatg atggtccg                                                   18 

 
           
             314  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            314 

ggaggagttc tggaacca                                                   18 

 
           
             315  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            315 

tgaggaagag gatgacgg                                                   18 

 
           
             316  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            316 

gcaggtcctt cttgttga                                                   18 

 
           
             317  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            317 

atcttgtcct ccagcagg                                                   18 

 
           
             318  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            318 

gcgagtacag gatcttgt                                                   18 

 
           
             319  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            319 

aagtagtcca ccaggtgc                                                   18 

 
           
             320  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            320 

gatgaactcc cgcgccgc                                                   18 

 
           
             321  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            321 

ggttcaggtc cacgaaca                                                   18 

 
           
             322  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            322 

gtagatgatc ttgtcgct                                                   18 

 
           
             323  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            323 

cacgtgaagt gtgagtag                                                   18 

 
           
             324  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            324 

atgttctccg tgtcggtg                                                   18 

 
           
             325  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            325 

acggccgcga acacgaag                                                   18 

 
           
             326  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            326 

gatggtgtcc ttcacggc                                                   18 

 
           
             327  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            327 

tcaggttcag ctgcagga                                                   18 

 
           
             328  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            328 

accagattgt actccttc                                                   18 

 
           
             329  
             2610  
             DNA  
             Homo sapiens  
             
               CDS  
               (199)...(1641)  
             
           
            329 

atcctgggac agggcacagg gccatctgtc accaggggct tagggaaggc cgagccagcc     60 

tgggtcaaag aagtcaaagg ggctgcctgg aggaggcagc ctgtcagctg gtgcatcaga    120 

ggctgtggcc aggccagctg ggctcgggga gcgccagcct gagaggagcg cgtgagcgtc    180 

gcgggagcct cgggcacc atg agc gac gtg gct att gtg aag gag ggt tgg      231 
                    Met Ser Asp Val Ala Ile Val Lys Glu Gly Trp 
                     1               5                   10 

ctg cac aaa cga ggg gag tac atc aag acc tgg cgg cca cgc tac ttc      279 
Leu His Lys Arg Gly Glu Tyr Ile Lys Thr Trp Arg Pro Arg Tyr Phe 
             15                  20                  25 

ctc ctc aag aat gat ggc acc ttc att ggc tac aag gag cgg ccg cag      327 
Leu Leu Lys Asn Asp Gly Thr Phe Ile Gly Tyr Lys Glu Arg Pro Gln 
         30                  35                  40 

gat gtg gac caa cgt gag gct ccc ctc aac aac ttc tct gtg gcg cag      375 
Asp Val Asp Gln Arg Glu Ala Pro Leu Asn Asn Phe Ser Val Ala Gln 
     45                  50                  55 

tgc cag ctg atg aag acg gag cgg ccc cgg ccc aac acc ttc atc atc      423 
Cys Gln Leu Met Lys Thr Glu Arg Pro Arg Pro Asn Thr Phe Ile Ile 
 60                  65                  70                  75 

cgc tgc ctg cag tgg acc act gtc atc gaa cgc acc ttc cat gtg gag      471 
Arg Cys Leu Gln Trp Thr Thr Val Ile Glu Arg Thr Phe His Val Glu 
                 80                  85                  90 

act cct gag gag cgg gag gag tgg aca acc gcc atc cag act gtg gct      519 
Thr Pro Glu Glu Arg Glu Glu Trp Thr Thr Ala Ile Gln Thr Val Ala 
             95                 100                 105 

gac ggc ctc aag aag cag gag gag gag gag atg gac ttc cgg tcg ggc      567 
Asp Gly Leu Lys Lys Gln Glu Glu Glu Glu Met Asp Phe Arg Ser Gly 
        110                 115                 120 

tca ccc agt gac aac tca ggg gct gaa gag atg gag gtg tcc ctg gcc      615 
Ser Pro Ser Asp Asn Ser Gly Ala Glu Glu Met Glu Val Ser Leu Ala 
    125                 130                 135 

aag ccc aag cac cgc gtg acc atg aac gag ttt gag tac ctg aag ctg      663 
Lys Pro Lys His Arg Val Thr Met Asn Glu Phe Glu Tyr Leu Lys Leu 
140                 145                 150                 155 

ctg ggc aag ggc act ttc ggc aag gtg atc ctg gtg aag gag aag gcc      711 
Leu Gly Lys Gly Thr Phe Gly Lys Val Ile Leu Val Lys Glu Lys Ala 
                160                 165                 170 

aca ggc cgc tac tac gcc atg aag atc ctc aag aag gaa gtc atc gtg      759 
Thr Gly Arg Tyr Tyr Ala Met Lys Ile Leu Lys Lys Glu Val Ile Val 
            175                 180                 185 

gcc aag gac gag gtg gcc cac aca ctc acc gag aac cgc gtc ctg cag      807 
Ala Lys Asp Glu Val Ala His Thr Leu Thr Glu Asn Arg Val Leu Gln 
        190                 195                 200 

aac tcc agg cac ccc ttc ctc aca gcc ctg aag tac tct ttc cag acc      855 
Asn Ser Arg His Pro Phe Leu Thr Ala Leu Lys Tyr Ser Phe Gln Thr 
    205                 210                 215 

cac gac cgc ctc tgc ttt gtc atg gag tac gcc aac ggg ggc gag ctg      903 
His Asp Arg Leu Cys Phe Val Met Glu Tyr Ala Asn Gly Gly Glu Leu 
220                 225                 230                 235 

ttc ttc cac ctg tcc cgg gaa cgt gtg ttc tcc gag gac cgg gcc cgc      951 
Phe Phe His Leu Ser Arg Glu Arg Val Phe Ser Glu Asp Arg Ala Arg 
                240                 245                 250 

ttc tat ggc gct gag att gtg tca gcc ctg gac tac ctg cac tcg gag      999 
Phe Tyr Gly Ala Glu Ile Val Ser Ala Leu Asp Tyr Leu His Ser Glu 
            255                 260                 265 

aag aac gtg gtg tac cgg gac ctc aag ctg gag aac ctc atg ctg gac     1047 
Lys Asn Val Val Tyr Arg Asp Leu Lys Leu Glu Asn Leu Met Leu Asp 
        270                 275                 280 

aag gac ggg cac att aag atc aca gac ttc ggg ctg tgc aag gag ggg     1095 
Lys Asp Gly His Ile Lys Ile Thr Asp Phe Gly Leu Cys Lys Glu Gly 
    285                 290                 295 

atc aag gac ggt gcc acc atg aag acc ttt tgc ggc aca cct gag tac     1143 
Ile Lys Asp Gly Ala Thr Met Lys Thr Phe Cys Gly Thr Pro Glu Tyr 
300                 305                 310                 315 

ctg gcc ccc gag gtg ctg gag gac aat gac tac ggc cgt gca gtg gac     1191 
Leu Ala Pro Glu Val Leu Glu Asp Asn Asp Tyr Gly Arg Ala Val Asp 
                320                 325                 330 

tgg tgg ggg ctg ggc gtg gtc atg tac gag atg atg tgc ggt cgc ctg     1239 
Trp Trp Gly Leu Gly Val Val Met Tyr Glu Met Met Cys Gly Arg Leu 
            335                 340                 345 

ccc ttc tac aac cag gac cat gag aag ctt ttt gag ctc atc ctc atg     1287 
Pro Phe Tyr Asn Gln Asp His Glu Lys Leu Phe Glu Leu Ile Leu Met 
        350                 355                 360 

gag gag atc cgc ttc ccg cgc acg ctt ggt ccc gag gcc aag tcc ttg     1335 
Glu Glu Ile Arg Phe Pro Arg Thr Leu Gly Pro Glu Ala Lys Ser Leu 
    365                 370                 375 

ctt tca ggg ctg ctc aag aag gac ccc aag cag agg ctt ggc ggg ggc     1383 
Leu Ser Gly Leu Leu Lys Lys Asp Pro Lys Gln Arg Leu Gly Gly Gly 
380                 385                 390                 395 

tcc gag gac gcc aag gag atc atg cag cat cgc ttc ttt gcc ggt atc     1431 
Ser Glu Asp Ala Lys Glu Ile Met Gln His Arg Phe Phe Ala Gly Ile 
                400                 405                 410 

gtg tgg cag cac gtg tac gag aag aag ctc agc cca ccc ttc aag ccc     1479 
Val Trp Gln His Val Tyr Glu Lys Lys Leu Ser Pro Pro Phe Lys Pro 
            415                 420                 425 

cag gtc acg tcg gag act gac acc agg tat ttt gat gag gag ttc acg     1527 
Gln Val Thr Ser Glu Thr Asp Thr Arg Tyr Phe Asp Glu Glu Phe Thr 
        430                 435                 440 

gcc cag atg atc acc atc aca cca cct gac caa gat gac agc atg gag     1575 
Ala Gln Met Ile Thr Ile Thr Pro Pro Asp Gln Asp Asp Ser Met Glu 
    445                 450                 455 

tgt gtg gac agc gag cgc agg ccc cac ttc ccc cag ttc tcc tac tcg     1623 
Cys Val Asp Ser Glu Arg Arg Pro His Phe Pro Gln Phe Ser Tyr Ser 
460                 465                 470                 475 

gcc agc agc acg gcc tga ggcggcggtg gactgcgctg gacgatagct            1671 
Ala Ser Ser Thr Ala  * 
                480 

tggagggatg gagaggcggc ctcgtgccat gatctgtatt taatggtttt tatttctcgg   1731 

gtgcatttga gagaagccac gctgtcctct cgagcccaga tggaaagacg tttttgtgct   1791 

gtgggcagca ccctcccccg cagcggggta gggaagaaaa ctatcctgcg ggttttaatt   1851 

tatttcatcc agtttgttct ccgggtgtgg cctcagccct cagaacaatc cgattcacgt   1911 

agggaaatgt taaggacttc tacagctatg cgcaatgtgg cattgggggg ccgggcaggt   1971 

cctgcccatg tgtcccctca ctctgtcagc cagccgccct gggctgtctg tcaccagcta   2031 

tctgtcatct ctctggggcc ctgggcctca gttcaacctg gtggcaccag atgcaacctc   2091 

actatggtat gctggccagc accctctcct gggggtggca ggcacacagc agccccccag   2151 

cactaaggcc gtgtctctga ggacgtcatc ggaggctggg cccctgggat gggaccaggg   2211 

atgggggatg ggccagggtt tacccagtgg gacagaggag caaggtttaa atttgttatt   2271 

gtgtattatg ttgttcaaat gcattttggg ggtttttaat ctttgtgaca ggaaagccct   2331 

cccccttccc cttctgtgtc acagttcttg gtgactgtcc caccggagcc tccccctcag   2391 

atgatctctc cacggtagca cttgaccttt tcgacgctta acctttccgc tgtcgcccca   2451 

ggccctccct gactccctgt gggggtggcc atccctgggc ccctccacgc ctcctggcca   2511 

gacgctgccg ctgccgctgc accacggcgt ttttttacaa cattcaactt tagtattttt   2571 

actattataa tataatatgg aaccttccct ccaaattct                          2610 

 
           
             330  
             21  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            330 

cgtgaccatg aacgagtttg a                                               21 

 
           
             331  
             19  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            331 

caggatcacc ttgccgaaa                                                  19 

 
           
             332  
             22  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            332 

ctgaagctgc tgggcaaggg ca                                              22 

 
           
             333  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            333 

ccctgtgccc tgtcccag                                                   18 

 
           
             334  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            334 

cctaagcccc tggtgaca                                                   18 

 
           
             335  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            335 

ctttgacttc tttgaccc                                                   18 

 
           
             336  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            336 

ggcagcccct ttgacttc                                                   18 

 
           
             337  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            337 

caaccctcct tcacaata                                                   18 

 
           
             338  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            338 

tactcccctc gtttgtgc                                                   18 

 
           
             339  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            339 

tgccatcatt cttgagga                                                   18 

 
           
             340  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            340 

agccaatgaa ggtgccat                                                   18 

 
           
             341  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            341 

cacagagaag ttgttgag                                                   18 

 
           
             342  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            342 

agtctggatg gcggttgt                                                   18 

 
           
             343  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            343 

tcctcctcct cctgcttc                                                   18 

 
           
             344  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            344 

cctgagttgt cactgggt                                                   18 

 
           
             345  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            345 

ccgaaagtgc ccttgccc                                                   18 

 
           
             346  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            346 

gccacgatga cttccttc                                                   18 

 
           
             347  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            347 

cggtcctcgg agaacaca                                                   18 

 
           
             348  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            348 

acgttcttct ccgagtgc                                                   18 

 
           
             349  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            349 

gtgccgcaaa aggtcttc                                                   18 

 
           
             350  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            350 

tactcaggtg tgccgcaa                                                   18 

 
           
             351  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            351 

ggcttgaagg gtgggctg                                                   18 

 
           
             352  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            352 

tcaaaatacc tggtgtca                                                   18 

 
           
             353  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            353 

gccgtgaact cctcatca                                                   18 

 
           
             354  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            354 

ggtcaggtgg tgtgatgg                                                   18 

 
           
             355  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            355 

ctcgctgtcc acacactc                                                   18 

 
           
             356  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            356 

gcctctccat ccctccaa                                                   18 

 
           
             357  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            357 

acagcgtggc ttctctca                                                   18 

 
           
             358  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            358 

ttttcttccc taccccgc                                                   18 

 
           
             359  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            359 

gatagttttc ttccctac                                                   18 

 
           
             360  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            360 

taaaacccgc aggatagt                                                   18 

 
           
             361  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            361 

ggagaacaaa ctggatga                                                   18 

 
           
             362  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            362 

ctggctgaca gagtgagg                                                   18 

 
           
             363  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            363 

gcggctggct gacagagt                                                   18 

 
           
             364  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            364 

cccagagaga tgacagat                                                   18 

 
           
             365  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            365 

gctgctgtgt gcctgcca                                                   18 

 
           
             366  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            366 

cataatacac aataacaa                                                   18 

 
           
             367  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            367 

atttgaacaa cataatac                                                   18 

 
           
             368  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            368 

aagtgctacc gtggagag                                                   18 

 
           
             369  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            369 

cgaaaaggtc aagtgcta                                                   18 

 
           
             370  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            370 

cagggagtca gggagggc                                                   18 

 
           
             371  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            371 

aaagttgaat gttgtaaa                                                   18 

 
           
             372  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            372 

aaaatactaa agttgaat                                                   18 

 
           
             373  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            373 

tccgtcatcg ctcctcaggg                                                 20 

 
           
             374  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            374 

gtgcgcgcga gcccgaaatc                                                 20 

 
           
             375  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            375 

atgcattctg cccccaagga                                                 20 

 
           
             376  
             4749  
             DNA  
             Homo sapiens  
             
               CDS  
               (52)...(3768)  
             
           
            376 

ggagcgggcg cgcggcggcg gcggggccgc ggcgggcggg tcgcgggggc a atg cgg      57 
                                                         Met Arg 
                                                          1 

gcg cag ggc cgg ggg gcc ttc ccc ccg gcg ctg ctg ctg ctg ctg gcg      105 
Ala Gln Gly Arg Gly Ala Phe Pro Pro Ala Leu Leu Leu Leu Leu Ala 
         5                   10                  15 

ctc tgg gtg cag gcg gcg cgg ccc atg ggc tat ttc gag ctg cag ctg      153 
Leu Trp Val Gln Ala Ala Arg Pro Met Gly Tyr Phe Glu Leu Gln Leu 
     20                  25                  30 

agc gcg ctg cgg aac gtg aac ggg gag ctg ctg agc ggc gcc tgc tgt      201 
Ser Ala Leu Arg Asn Val Asn Gly Glu Leu Leu Ser Gly Ala Cys Cys 
 35                  40                  45                  50 

gac ggc gac ggc cgg aca acg cgc gcg ggg ggc tgc ggc cac gac gag      249 
Asp Gly Asp Gly Arg Thr Thr Arg Ala Gly Gly Cys Gly His Asp Glu 
                 55                  60                  65 

tgc gac acg tac gtg cgc gtg tgc ctt aag gag tac cag gcc aag gtg      297 
Cys Asp Thr Tyr Val Arg Val Cys Leu Lys Glu Tyr Gln Ala Lys Val 
             70                  75                  80 

acg ccc acg ggg ccc tgc agc tac ggc cac ggc gcc acg ccc gtg ctg      345 
Thr Pro Thr Gly Pro Cys Ser Tyr Gly His Gly Ala Thr Pro Val Leu 
         85                  90                  95 

ggc ggc aac tcc ttc tac ctg ccg ccg gcg ggc gct gcg ggg gac cga      393 
Gly Gly Asn Ser Phe Tyr Leu Pro Pro Ala Gly Ala Ala Gly Asp Arg 
    100                 105                 110 

gcg cgc gcg cgg ccc cgg gcc ggc ggc gac cag gac ccg ggc ttc gtc      441 
Ala Arg Ala Arg Pro Arg Ala Gly Gly Asp Gln Asp Pro Gly Phe Val 
115                 120                 125                 130 

gtc atc ccc ttc cag ttc gcc tgg ccg cgc tcc ttt acc ctc atc gtg      489 
Val Ile Pro Phe Gln Phe Ala Trp Pro Arg Ser Phe Thr Leu Ile Val 
                135                 140                 145 

gag gcc tgg gac tgg gac aac gat acc acc ccg aat gag gag ctg ctg      537 
Glu Ala Trp Asp Trp Asp Asn Asp Thr Thr Pro Asn Glu Glu Leu Leu 
            150                 155                 160 

atc gag cga gtg tcg cat gcc ggc atg atc aac ccg gag gac cgc tgg      585 
Ile Glu Arg Val Ser His Ala Gly Met Ile Asn Pro Glu Asp Arg Trp 
        165                 170                 175 

aag agc ctg cac ttc agc ggc cac gtg gcg cac ctg gag ctg cag atc      633 
Lys Ser Leu His Phe Ser Gly His Val Ala His Leu Glu Leu Gln Ile 
    180                 185                 190 

cgc gtg cgc tgc gac gag aac tac tac agc gcc act tgc aac aag ttc      681 
Arg Val Arg Cys Asp Glu Asn Tyr Tyr Ser Ala Thr Cys Asn Lys Phe 
195                 200                 205                 210 

tgc cgg ccc cgc aac gac ttt ttc ggc cac tac acc tgc gac cag tac      729 
Cys Arg Pro Arg Asn Asp Phe Phe Gly His Tyr Thr Cys Asp Gln Tyr 
                215                 220                 225 

ggc aac aag gcc tgc atg gac ggc tgg atg ggc aag gag tgc aag gaa      777 
Gly Asn Lys Ala Cys Met Asp Gly Trp Met Gly Lys Glu Cys Lys Glu 
            230                 235                 240 

gct gtg tgt aaa caa ggg tgt aat ttg ctc cac ggg gga tgc acc gtg      825 
Ala Val Cys Lys Gln Gly Cys Asn Leu Leu His Gly Gly Cys Thr Val 
        245                 250                 255 

cct ggg gag tgc agg tgc agc tac ggc tgg caa ggg agg ttc tgc gat      873 
Pro Gly Glu Cys Arg Cys Ser Tyr Gly Trp Gln Gly Arg Phe Cys Asp 
    260                 265                 270 

gag tgt gtc ccc tac ccc ggc tgc gtg cat ggc agt tgt gtg gag ccc      921 
Glu Cys Val Pro Tyr Pro Gly Cys Val His Gly Ser Cys Val Glu Pro 
275                 280                 285                 290 

tgg cag tgc aac tgt gag acc aac tgg ggc ggc ctg ctc tgt gac aaa      969 
Trp Gln Cys Asn Cys Glu Thr Asn Trp Gly Gly Leu Leu Cys Asp Lys 
                295                 300                 305 

gac ctg aac tac tgt ggc agc cac cac ccc tgc acc aac gga ggc acg     1017 
Asp Leu Asn Tyr Cys Gly Ser His His Pro Cys Thr Asn Gly Gly Thr 
            310                 315                 320 

tgc atc aac gcc gag cct gac cag tac cgc tgc acc tgc cct gac ggc     1065 
Cys Ile Asn Ala Glu Pro Asp Gln Tyr Arg Cys Thr Cys Pro Asp Gly 
        325                 330                 335 

tac tcg ggc agg aac tgt gag aag gct gag cac gcc tgc acc tcc aac     1113 
Tyr Ser Gly Arg Asn Cys Glu Lys Ala Glu His Ala Cys Thr Ser Asn 
    340                 345                 350 

ccg tgt gcc aac ggg ggc tct tgc cat gag gtg ccg tcc ggc ttc gaa     1161 
Pro Cys Ala Asn Gly Gly Ser Cys His Glu Val Pro Ser Gly Phe Glu 
355                 360                 365                 370 

tgc cac tgc cca tcg ggc tgg agc ggg ccc acc tgt gcc ctt gac atc     1209 
Cys His Cys Pro Ser Gly Trp Ser Gly Pro Thr Cys Ala Leu Asp Ile 
                375                 380                 385 

gat gag tgt gct tcg aac ccg tgt gcg gcc ggt ggc acc tgt gtg gac     1257 
Asp Glu Cys Ala Ser Asn Pro Cys Ala Ala Gly Gly Thr Cys Val Asp 
            390                 395                 400 

cag gtg gac ggc ttt gag tgc atc tgc ccc gag cag tgg gtg ggg gcc     1305 
Gln Val Asp Gly Phe Glu Cys Ile Cys Pro Glu Gln Trp Val Gly Ala 
        405                 410                 415 

acc tgc cag ctg gac gtc aac gac tgt gaa ggg aag cca tgc ctt aac     1353 
Thr Cys Gln Leu Asp Val Asn Asp Cys Glu Gly Lys Pro Cys Leu Asn 
    420                 425                 430 

gct ttt tct tgc aaa aac ctg att ggc ggc tat tac tgt gat tgc atc     1401 
Ala Phe Ser Cys Lys Asn Leu Ile Gly Gly Tyr Tyr Cys Asp Cys Ile 
435                 440                 445                 450 

ccg ggc tgg aag ggc atc aac tgc cat atc aac gtc aac gac tgt cgc     1449 
Pro Gly Trp Lys Gly Ile Asn Cys His Ile Asn Val Asn Asp Cys Arg 
                455                 460                 465 

ggg cag tgt cag cat ggg ggc acc tgc aag gac ctg gtg aac ggg tac     1497 
Gly Gln Cys Gln His Gly Gly Thr Cys Lys Asp Leu Val Asn Gly Tyr 
            470                 475                 480 

cag tgt gtg tgc cca cgg ggc ttc gga ggc cgg cat tgc gag ctg gaa     1545 
Gln Cys Val Cys Pro Arg Gly Phe Gly Gly Arg His Cys Glu Leu Glu 
        485                 490                 495 

cga gac aag tgt gcc agc agc ccc tgc cac agc ggc ggc ctc tgc gag     1593 
Arg Asp Lys Cys Ala Ser Ser Pro Cys His Ser Gly Gly Leu Cys Glu 
    500                 505                 510 

gac ctg gcc gac ggc ttc cac tgc cac tgc ccc cag ggc ttc tcc ggg     1641 
Asp Leu Ala Asp Gly Phe His Cys His Cys Pro Gln Gly Phe Ser Gly 
515                 520                 525                 530 

cct ctc tgt gag gtg gat gtc gac ctt tgt gag cca agc ccc tgc cgg     1689 
Pro Leu Cys Glu Val Asp Val Asp Leu Cys Glu Pro Ser Pro Cys Arg 
                535                 540                 545 

aac ggc gct cgc tgc tat aac ctg gag ggt gac tat tac tgc gcc tgc     1737 
Asn Gly Ala Arg Cys Tyr Asn Leu Glu Gly Asp Tyr Tyr Cys Ala Cys 
            550                 555                 560 

cct gat gac ttt ggt ggc aag aac tgc tcc gtg ccc cgc gag ccg tgc     1785 
Pro Asp Asp Phe Gly Gly Lys Asn Cys Ser Val Pro Arg Glu Pro Cys 
        565                 570                 575 

cct ggc ggg gcc tgc aga gtg atc gat ggc tgc ggg tca gac gcg ggg     1833 
Pro Gly Gly Ala Cys Arg Val Ile Asp Gly Cys Gly Ser Asp Ala Gly 
    580                 585                 590 

cct ggg atg cct ggc aca gca gcc tcc ggc gtg tgt ggc ccc cat gga     1881 
Pro Gly Met Pro Gly Thr Ala Ala Ser Gly Val Cys Gly Pro His Gly 
595                 600                 605                 610 

cgc tgc gtc agc cag cca ggg ggc aac ttt tcc tgc atc tgt gac agt     1929 
Arg Cys Val Ser Gln Pro Gly Gly Asn Phe Ser Cys Ile Cys Asp Ser 
                615                 620                 625 

ggc ttt act ggc acc tac tgc cat gag aac att gac gac tgc ctg ggc     1977 
Gly Phe Thr Gly Thr Tyr Cys His Glu Asn Ile Asp Asp Cys Leu Gly 
            630                 635                 640 

cag ccc tgc cgc aat ggg ggc aca tgc atc gat gag gtg gac gcc ttc     2025 
Gln Pro Cys Arg Asn Gly Gly Thr Cys Ile Asp Glu Val Asp Ala Phe 
        645                 650                 655 

cgc tgc ttc tgc ccc agc ggc tgg gag ggc gag ctc tgc gac acc aat     2073 
Arg Cys Phe Cys Pro Ser Gly Trp Glu Gly Glu Leu Cys Asp Thr Asn 
    660                 665                 670 

ccc aac gac tgc ctt ccc gat ccc tgc cac agc cgc ggc cgc tgc tac     2121 
Pro Asn Asp Cys Leu Pro Asp Pro Cys His Ser Arg Gly Arg Cys Tyr 
675                 680                 685                 690 

gac ctg gtc aat gac ttc tac tgt gcg tgc gac gac ggc tgg aag ggc     2169 
Asp Leu Val Asn Asp Phe Tyr Cys Ala Cys Asp Asp Gly Trp Lys Gly 
                695                 700                 705 

aag acc tgc cac tca cgc gag ttc cag tgc gat gcc tac acc tgc agc     2217 
Lys Thr Cys His Ser Arg Glu Phe Gln Cys Asp Ala Tyr Thr Cys Ser 
            710                 715                 720 

aac ggt ggc acc tgc tac gac agc ggc gac acc ttc cgc tgc gcc tgc     2265 
Asn Gly Gly Thr Cys Tyr Asp Ser Gly Asp Thr Phe Arg Cys Ala Cys 
        725                 730                 735 

ccc ccc ggc tgg aag ggc agc acc tgc gcc gtc gcc aag aac agc agc     2313 
Pro Pro Gly Trp Lys Gly Ser Thr Cys Ala Val Ala Lys Asn Ser Ser 
    740                 745                 750 

tgc ctg ccc aac ccc tgt gtg aat ggt ggc acc tgc gtg ggc agc ggg     2361 
Cys Leu Pro Asn Pro Cys Val Asn Gly Gly Thr Cys Val Gly Ser Gly 
755                 760                 765                 770 

gcc tcc ttc tcc tgc atc tgc cgg gac ggc tgg gag ggt cgt act tgc     2409 
Ala Ser Phe Ser Cys Ile Cys Arg Asp Gly Trp Glu Gly Arg Thr Cys 
                775                 780                 785 

act cac aat acc aac gac tgc aac cct ctg cct tgc tac aat ggt ggc     2457 
Thr His Asn Thr Asn Asp Cys Asn Pro Leu Pro Cys Tyr Asn Gly Gly 
            790                 795                 800 

atc tgt gtt gac ggc gtc aac tgg ttc cgc tgc gag tgt gca cct ggc     2505 
Ile Cys Val Asp Gly Val Asn Trp Phe Arg Cys Glu Cys Ala Pro Gly 
        805                 810                 815 

ttc gcg ggg cct gac tgc cgc atc aac atc gac gag tgc cag tcc tcg     2553 
Phe Ala Gly Pro Asp Cys Arg Ile Asn Ile Asp Glu Cys Gln Ser Ser 
    820                 825                 830 

ccc tgt gcc tac ggg gcc acg tgt gtg gat gag atc aac ggg tat cgc     2601 
Pro Cys Ala Tyr Gly Ala Thr Cys Val Asp Glu Ile Asn Gly Tyr Arg 
835                 840                 845                 850 

tgt agc tgc cca ccc ggc cga gcc ggc ccc cgg tgc cag gaa gtg atc     2649 
Cys Ser Cys Pro Pro Gly Arg Ala Gly Pro Arg Cys Gln Glu Val Ile 
                855                 860                 865 

ggg ttc ggg aga tcc tgc tgg tcc cgg ggc act ccg ttc cca cac gga     2697 
Gly Phe Gly Arg Ser Cys Trp Ser Arg Gly Thr Pro Phe Pro His Gly 
            870                 875                 880 

agc tcc tgg gtg gaa gac tgc aac agc tgc cgc tgc ctg gat ggc cgc     2745 
Ser Ser Trp Val Glu Asp Cys Asn Ser Cys Arg Cys Leu Asp Gly Arg 
        885                 890                 895 

cgt gac tgc agc aag gtg tgg tgc gga tgg aag cct tgt ctg ctg gcc     2793 
Arg Asp Cys Ser Lys Val Trp Cys Gly Trp Lys Pro Cys Leu Leu Ala 
    900                 905                 910 

ggc cag ccc gag gcc ctg agc gcc cag tgc cca ctg ggg caa agg tgc     2841 
Gly Gln Pro Glu Ala Leu Ser Ala Gln Cys Pro Leu Gly Gln Arg Cys 
915                 920                 925                 930 

ctg gag aag gcc cca ggc cag tgt ctg cga cca ccc tgt gag gcc tgg     2889 
Leu Glu Lys Ala Pro Gly Gln Cys Leu Arg Pro Pro Cys Glu Ala Trp 
                935                 940                 945 

ggg gag tgc ggc gca gaa gag cca ccg agc acc ccc tgc ctg cca cgc     2937 
Gly Glu Cys Gly Ala Glu Glu Pro Pro Ser Thr Pro Cys Leu Pro Arg 
            950                 955                 960 

tcc ggc cac ctg gac aat aac tgt gcc cgc ctc acc ttg cat ttc aac     2985 
Ser Gly His Leu Asp Asn Asn Cys Ala Arg Leu Thr Leu His Phe Asn 
        965                 970                 975 

cgt gac cac gtg ccc cag ggc acc acg gtg ggc gcc att tgc tcc ggg     3033 
Arg Asp His Val Pro Gln Gly Thr Thr Val Gly Ala Ile Cys Ser Gly 
    980                 985                 990 

atc cgc tcc ctg cca gcc aca agg gct gtg gca cgg gac cgc ctg ctg     3081 
Ile Arg Ser Leu Pro Ala Thr Arg Ala Val Ala Arg Asp Arg Leu Leu 
 995                1000                1005                1010 

gtg ttg ctt tgc gac cgg gcg tcc tcg ggg gcc agt gcc gtg gag gtg     3129 
Val Leu Leu Cys Asp Arg Ala Ser Ser Gly Ala Ser Ala Val Glu Val 
                1015                1020                1025 

gcc gtg tcc ttc agc cct gcc agg gac ctg cct gac agc agc ctg atc     3177 
Ala Val Ser Phe Ser Pro Ala Arg Asp Leu Pro Asp Ser Ser Leu Ile 
            1030                1035                1040 

cag ggc gcg gcc cac gcc atc gtg gcc gcc atc acc cag cgg ggg aac     3225 
Gln Gly Ala Ala His Ala Ile Val Ala Ala Ile Thr Gln Arg Gly Asn 
        1045                1050                1055 

agc tca ctg ctc ctg gct gtc acc gag gtc aag gtg gag acg gtt gtt     3273 
Ser Ser Leu Leu Leu Ala Val Thr Glu Val Lys Val Glu Thr Val Val 
    1060                1065                1070 

acg ggc ggc tct tcc aca ggt ctg ctg gtg cct gtg ctg tgt ggt gcc     3321 
Thr Gly Gly Ser Ser Thr Gly Leu Leu Val Pro Val Leu Cys Gly Ala 
1075                1080                1085                1090 

ttc agc gtg ctg tgg ctg gcg tgc gtg gtc ctg tgc gtg tgg tgg aca     3369 
Phe Ser Val Leu Trp Leu Ala Cys Val Val Leu Cys Val Trp Trp Thr 
                1095                1100                1105 

cgc aag cgc agg aaa gag cgg gag agg agc cgg ctg ccg cgg gag gag     3417 
Arg Lys Arg Arg Lys Glu Arg Glu Arg Ser Arg Leu Pro Arg Glu Glu 
            1110                1115                1120 

agc gcc aac aac cag tgg gcc ccg ctc aac ccc atc cgc aac ccc atc     3465 
Ser Ala Asn Asn Gln Trp Ala Pro Leu Asn Pro Ile Arg Asn Pro Ile 
        1125                1130                1135 

gag cgg ccg ggg ggc cac aag gac gtg ctc tac cag tgc aag aac ttc     3513 
Glu Arg Pro Gly Gly His Lys Asp Val Leu Tyr Gln Cys Lys Asn Phe 
    1140                1145                1150 

acg ccg ccg ccg cgc agg gcg gac gag gcg ctg ccc ggg ccg gcc ggc     3561 
Thr Pro Pro Pro Arg Arg Ala Asp Glu Ala Leu Pro Gly Pro Ala Gly 
1155                1160                1165                1170 

cac gcg gcc gtc agg gag gat gag gag gac gag gat ctg ggc cgc ggt     3609 
His Ala Ala Val Arg Glu Asp Glu Glu Asp Glu Asp Leu Gly Arg Gly 
                1175                1180                1185 

gag gag gac tcc ctg gag gcg gag aag ttc ctc tca cac aaa ttc acc     3657 
Glu Glu Asp Ser Leu Glu Ala Glu Lys Phe Leu Ser His Lys Phe Thr 
            1190                1195                1200 

aaa gat cct ggc cgc tcg ccg ggg agg ccg gcc cac tgg gcc tca ggc     3705 
Lys Asp Pro Gly Arg Ser Pro Gly Arg Pro Ala His Trp Ala Ser Gly 
        1205                1210                1215 

ccc aaa gtg gac aac cgc gcg gtc agg agc atc aat gag gcc cgc tac     3753 
Pro Lys Val Asp Asn Arg Ala Val Arg Ser Ile Asn Glu Ala Arg Tyr 
    1220                1225                1230 

gcc ggc aag gag tag gggcggctgc agctgggccg ggacccaggg ccctcggtgg     3808 
Ala Gly Lys Glu  * 
1235 

gagccatgcc gtctgccgga cccggagccg aggcatgtgc atagtttctt tattttgtgt   3868 

aaaaaaacca ccaaaaacaa aaaccaaatg tttattttct acgtttcttt aaccttgtat   3928 

aaattattca gtaactgtca ggctgaaaac aatggagtat tctcggatag ttgctatttt   3988 

tgtaaagttt ccgtgcgtgg cactcgctgt atgaaaggag agagcaaagg gtgtctgcgt   4048 

cgtcaccaaa tcgtagcgtt tgttaccaga ggttgtgcac tgtttacaga atcttccttt   4108 

tattcctcac tcgggtttct ctgtggctcc aggccaaagt gccggtgaga cccatggctg   4168 

tgttggtgtg gcccatggct gttggtggga cccgtggctg atggtgtggc ctgtggctgt   4228 

cggtgggact cgtggctgtc aatgggacct gtggctgtcg gtgggaccta cggtggtcgg   4288 

tgggaccctg gttattgatg tggccctggc tgccggcacg gcccgtggct gttgacgcac   4348 

ctgtggttgt tagtggggcc tgaggtcatc ggcgtgccca aggccggcag gtcaacctcg   4408 

cgcttgctgg ccagtccacc ctgcctgccg tctgtgcttc ctcctgccca gaacgcccgc   4468 

tccagcgatc tctccactgt gctttcagaa gtgcccttcc tgctgcgcag ttctcccatc   4528 

ctgggacggc ggcagtattg aagctcgtga caagtgcctt cacacagacc cctcgcaact   4588 

gtccacgcgt gccgtggcac caggcgctgc ccacctgccg gccccggccg cccctcctcg   4648 

tgaaagtgca tttttgtaaa tgtgtacata ttaaaggaag cactctgtat atttgattga   4708 

ataatgccac caaaaaaaaa aaaaaaaaaa aattcctgcc c                       4749 

 
           
             377  
             16  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            377 

cccagggctt ctccgg                                                     16 

 
           
             378  
             26  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            378 

aatagtcacc ctccaggtta tagcag                                          26 

 
           
             379  
             26  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            379 

tggatgtcga cctttgtgag ccaagc                                          26 

 
           
             380  
             4974  
             DNA  
             Homo sapiens  
             
               CDS  
               (405)...(4121)  
             
           
            380 

ctcatgcata tgcaggtgcg cgggtgacga atgggcgagc gagctgtcag tctcgttccg     60 

aacttgttgg ctgcggtgcc gggagcgcgg gcgcgcagag cccgaggccg ggacccgctg    120 

ccttcaccgc cgccgccgtc gccgccgggt gggagccggg ccgggcagcc ggagcgcggc    180 

cgccagcgag ccggagctgc cgccgcccct gcacgcccgc cgcccaggcc cgcgcgccgg    240 

acgctgcgct cgaccccgcc cgcgccgccg ccgccgccgc ctctgccgct gccgctgcct    300 

ctgcgggcgc tcggagggcg ggcgggcgct gggaggccgg cgcggcggct gggagccggg    360 

cgcgggcggc ggcggcgggg ccgggcgggc gggtcgcggg ggca atg cgg gcg cag     416 
                                                 Met Arg Ala Gln 
                                                  1 

ggc cgg ggg cgc ctt ccc cgg cgg ctg ctg ctg ctg ctg gcg ctc tgg      464 
Gly Arg Gly Arg Leu Pro Arg Arg Leu Leu Leu Leu Leu Ala Leu Trp 
 5                   10                  15                  20 

gtg cag gcg gcg cgg ccc atg ggc tat ttc gag ctg cag ctg agc gcg      512 
Val Gln Ala Ala Arg Pro Met Gly Tyr Phe Glu Leu Gln Leu Ser Ala 
                 25                  30                  35 

ctg cgg aac gtg aac ggg gag ctg ctg agc ggc gcc tgc tgt gac ggc      560 
Leu Arg Asn Val Asn Gly Glu Leu Leu Ser Gly Ala Cys Cys Asp Gly 
             40                  45                  50 

gac ggc cgg aca acg cgc gcg ggg ggc tgc ggc cac gac gag tgc gac      608 
Asp Gly Arg Thr Thr Arg Ala Gly Gly Cys Gly His Asp Glu Cys Asp 
         55                  60                  65 

acg tac gtg cgc gtg tgc ctt aag gag tac cag gcc aag gtg acg ccc      656 
Thr Tyr Val Arg Val Cys Leu Lys Glu Tyr Gln Ala Lys Val Thr Pro 
     70                  75                  80 

acg ggg ccc tgc agc tac ggc cac ggc gcc acg ccc gtg ctg ggc ggc      704 
Thr Gly Pro Cys Ser Tyr Gly His Gly Ala Thr Pro Val Leu Gly Gly 
 85                  90                  95                 100 

aac tcc ttc tac ctg ccg ccg gcg ggc gct gcg ggg gac cga gcg cgg      752 
Asn Ser Phe Tyr Leu Pro Pro Ala Gly Ala Ala Gly Asp Arg Ala Arg 
                105                 110                 115 

gcg cgg gcc cgg gcc ggc ggc gac cag gac ccg ggc ctc gtc gtc atc      800 
Ala Arg Ala Arg Ala Gly Gly Asp Gln Asp Pro Gly Leu Val Val Ile 
            120                 125                 130 

ccc ttc cag ttc gcc tgg ccg cgc tcc ttt acc ctc atc gtg gag gcc      848 
Pro Phe Gln Phe Ala Trp Pro Arg Ser Phe Thr Leu Ile Val Glu Ala 
        135                 140                 145 

tgg gac tgg gac aac gat acc acc ccg aat gag gag ctg ctg atc gag      896 
Trp Asp Trp Asp Asn Asp Thr Thr Pro Asn Glu Glu Leu Leu Ile Glu 
    150                 155                 160 

cga gtg tcg cat gcc ggc atg atc aac ccg gag gac cgc tgg aag agc      944 
Arg Val Ser His Ala Gly Met Ile Asn Pro Glu Asp Arg Trp Lys Ser 
165                 170                 175                 180 

ctg cac ttc agc ggc cac gtg gcg cac ctg gag ctg cag atc cgc gtg      992 
Leu His Phe Ser Gly His Val Ala His Leu Glu Leu Gln Ile Arg Val 
                185                 190                 195 

cgc tgc gac gag aac tac tac agc gcc act tgc aac aag ttc tgc cgg     1040 
Arg Cys Asp Glu Asn Tyr Tyr Ser Ala Thr Cys Asn Lys Phe Cys Arg 
            200                 205                 210 

ccc cgc aac gac ttt ttc ggc cac tac acc tgc gac cag tac ggc aac     1088 
Pro Arg Asn Asp Phe Phe Gly His Tyr Thr Cys Asp Gln Tyr Gly Asn 
        215                 220                 225 

aag gcc tgc atg gac ggc tgg atg ggc aag gag tgc aag gaa gct gtg     1136 
Lys Ala Cys Met Asp Gly Trp Met Gly Lys Glu Cys Lys Glu Ala Val 
    230                 235                 240 

tgt aaa caa ggg tgt aat ttg ctc cac ggg gga tgc acc gtg cct ggg     1184 
Cys Lys Gln Gly Cys Asn Leu Leu His Gly Gly Cys Thr Val Pro Gly 
245                 250                 255                 260 

gag tgc agg tgc agc tac ggc tgg caa ggg agg ttc tgc gat gag tgt     1232 
Glu Cys Arg Cys Ser Tyr Gly Trp Gln Gly Arg Phe Cys Asp Glu Cys 
                265                 270                 275 

gtc ccc tac ccc ggc tgc gtg cat ggc agt tgt gtg gag ccc tgg cag     1280 
Val Pro Tyr Pro Gly Cys Val His Gly Ser Cys Val Glu Pro Trp Gln 
            280                 285                 290 

tgc aac tgt gag acc aac tgg ggc ggc ctg ctc tgt gac aaa gac ctg     1328 
Cys Asn Cys Glu Thr Asn Trp Gly Gly Leu Leu Cys Asp Lys Asp Leu 
        295                 300                 305 

aac tac tgt ggc agc cac cac ccc tgc acc aac gga ggc acg tgc atc     1376 
Asn Tyr Cys Gly Ser His His Pro Cys Thr Asn Gly Gly Thr Cys Ile 
    310                 315                 320 

aac gcc gag cct gac cag tac cgc tgc acc tgc cct gac ggc tac tcg     1424 
Asn Ala Glu Pro Asp Gln Tyr Arg Cys Thr Cys Pro Asp Gly Tyr Ser 
325                 330                 335                 340 

ggc agg aac tgt gag aag gct gag cac gcc tgc acc tcc aac ccg tgt     1472 
Gly Arg Asn Cys Glu Lys Ala Glu His Ala Cys Thr Ser Asn Pro Cys 
                345                 350                 355 

gcc aac ggg ggc tct tgc cat gag gtg ccg tcc ggc ttc gaa tgc cac     1520 
Ala Asn Gly Gly Ser Cys His Glu Val Pro Ser Gly Phe Glu Cys His 
            360                 365                 370 

tgc cca tcg ggc tgg agc ggg ccc acc tgt gcc ctt gac atc gat gag     1568 
Cys Pro Ser Gly Trp Ser Gly Pro Thr Cys Ala Leu Asp Ile Asp Glu 
        375                 380                 385 

tgt gct tcg aac ccg tgt gcg gcc ggt ggc acc tgt gtg gac cag gtg     1616 
Cys Ala Ser Asn Pro Cys Ala Ala Gly Gly Thr Cys Val Asp Gln Val 
    390                 395                 400 

gac ggc ttt gag tgc atc tgc ccc gag cag tgg gtg ggg gcc acc tgc     1664 
Asp Gly Phe Glu Cys Ile Cys Pro Glu Gln Trp Val Gly Ala Thr Cys 
405                 410                 415                 420 

cag ctg gac gcc aat gag tgt gaa ggg aag cca tgc ctt aac gct ttt     1712 
Gln Leu Asp Ala Asn Glu Cys Glu Gly Lys Pro Cys Leu Asn Ala Phe 
                425                 430                 435 

tct tgc aaa aac ctg att ggc ggc tat tac tgt gat tgc atc ccg ggc     1760 
Ser Cys Lys Asn Leu Ile Gly Gly Tyr Tyr Cys Asp Cys Ile Pro Gly 
            440                 445                 450 

tgg aag ggc atc aac tgc cat atc aac gtc aac gac tgt cgc ggg cag     1808 
Trp Lys Gly Ile Asn Cys His Ile Asn Val Asn Asp Cys Arg Gly Gln 
        455                 460                 465 

tgt cag cat ggg ggc acc tgc aag gac ctg gtg aac ggg tac cag tgt     1856 
Cys Gln His Gly Gly Thr Cys Lys Asp Leu Val Asn Gly Tyr Gln Cys 
    470                 475                 480 

gtg tgc cca cgg ggc ttc gga ggc cgg cat tgc gag ctg gaa cga gac     1904 
Val Cys Pro Arg Gly Phe Gly Gly Arg His Cys Glu Leu Glu Arg Asp 
485                 490                 495                 500 

aag tgt gcc agc agc ccc tgc cac agc ggc ggc ctc tgc gag gac ctg     1952 
Lys Cys Ala Ser Ser Pro Cys His Ser Gly Gly Leu Cys Glu Asp Leu 
                505                 510                 515 

gcc gac ggc ttc cac tgc cac tgc ccc cag ggc ttc tcc ggg cct ctc     2000 
Ala Asp Gly Phe His Cys His Cys Pro Gln Gly Phe Ser Gly Pro Leu 
            520                 525                 530 

tgt gag gtg gat gtc gac ctt tgt gag cca agc ccc tgc cgg aac ggc     2048 
Cys Glu Val Asp Val Asp Leu Cys Glu Pro Ser Pro Cys Arg Asn Gly 
        535                 540                 545 

gct cgc tgc tat aac ctg gag ggt gac tat tac tgc gcc tgc cct gat     2096 
Ala Arg Cys Tyr Asn Leu Glu Gly Asp Tyr Tyr Cys Ala Cys Pro Asp 
    550                 555                 560 

gac ttt ggt ggc aag aac tgc tcc gtg ccc cgc gag ccg tgc cct ggc     2144 
Asp Phe Gly Gly Lys Asn Cys Ser Val Pro Arg Glu Pro Cys Pro Gly 
565                 570                 575                 580 

ggg gcc tgc aga gtg atc gat ggc tgc ggg tca gac gcg ggg cct ggg     2192 
Gly Ala Cys Arg Val Ile Asp Gly Cys Gly Ser Asp Ala Gly Pro Gly 
                585                 590                 595 

atg cct ggc aca gca gcc tcc ggc gtg tgt ggc ccc cat gga cgc tgc     2240 
Met Pro Gly Thr Ala Ala Ser Gly Val Cys Gly Pro His Gly Arg Cys 
            600                 605                 610 

gtc agc cag cca ggg ggc aac ttt tcc tgc atc tgt gac agt ggc ttt     2288 
Val Ser Gln Pro Gly Gly Asn Phe Ser Cys Ile Cys Asp Ser Gly Phe 
        615                 620                 625 

act ggc acc tac tgc cat gag aac att gac gac tgc ctg ggc cag ccc     2336 
Thr Gly Thr Tyr Cys His Glu Asn Ile Asp Asp Cys Leu Gly Gln Pro 
    630                 635                 640 

tgc cgc aat ggg ggc aca tgc atc gat gag gtg gac gcc ttc cgc tgc     2384 
Cys Arg Asn Gly Gly Thr Cys Ile Asp Glu Val Asp Ala Phe Arg Cys 
645                 650                 655                 660 

ttc tgc ccc agc ggc tgg gag ggc gag ctc tgc gac acc aat ccc aac     2432 
Phe Cys Pro Ser Gly Trp Glu Gly Glu Leu Cys Asp Thr Asn Pro Asn 
                665                 670                 675 

gac tgc ctt ccc gat ccc tgc cac agc cgc ggc cgc tgc tac gac ctg     2480 
Asp Cys Leu Pro Asp Pro Cys His Ser Arg Gly Arg Cys Tyr Asp Leu 
            680                 685                 690 

gtc aat gac ttc tac tgt gcg tgc gac gac ggc tgg aag ggc aag acc     2528 
Val Asn Asp Phe Tyr Cys Ala Cys Asp Asp Gly Trp Lys Gly Lys Thr 
        695                 700                 705 

tgc cac tca cgc gag ttc cag tgc gat gcc tac acc tgc agc aac ggt     2576 
Cys His Ser Arg Glu Phe Gln Cys Asp Ala Tyr Thr Cys Ser Asn Gly 
    710                 715                 720 

ggc acc tgc tac gac agc ggc gac acc ttc cgc tgc gcc tgc ccc ccc     2624 
Gly Thr Cys Tyr Asp Ser Gly Asp Thr Phe Arg Cys Ala Cys Pro Pro 
725                 730                 735                 740 

ggc tgg aag ggc agc acc tgc gcc gtc gcc aag aac agc agc tgc ctg     2672 
Gly Trp Lys Gly Ser Thr Cys Ala Val Ala Lys Asn Ser Ser Cys Leu 
                745                 750                 755 

ccc aac ccc tgt gtg aat ggt ggc acc tgc gtg ggc agc ggg gcc tcc     2720 
Pro Asn Pro Cys Val Asn Gly Gly Thr Cys Val Gly Ser Gly Ala Ser 
            760                 765                 770 

ttc tcc tgc atc tgc cgg gac ggc tgg gag ggt cgt act tgc act cac     2768 
Phe Ser Cys Ile Cys Arg Asp Gly Trp Glu Gly Arg Thr Cys Thr His 
        775                 780                 785 

aat acc aac gac tgc aac cct ctg cct tgc tac aat ggt ggc atc tgt     2816 
Asn Thr Asn Asp Cys Asn Pro Leu Pro Cys Tyr Asn Gly Gly Ile Cys 
    790                 795                 800 

gtt gac ggc gtc aac tgg ttc cgc tgc gag tgt gca cct ggc ttc gcg     2864 
Val Asp Gly Val Asn Trp Phe Arg Cys Glu Cys Ala Pro Gly Phe Ala 
805                 810                 815                 820 

ggg cct gac tgc cgc atc aac atc gac gag tgc cag tcc tcg ccc tgt     2912 
Gly Pro Asp Cys Arg Ile Asn Ile Asp Glu Cys Gln Ser Ser Pro Cys 
                825                 830                 835 

gcc tac ggg gcc acg tgt gtg gat gag atc aac ggg tat cgc tgt agc     2960 
Ala Tyr Gly Ala Thr Cys Val Asp Glu Ile Asn Gly Tyr Arg Cys Ser 
            840                 845                 850 

tgc cca ccc ggc cga gcc ggc ccc cgg tgc cag gaa gtg atc ggg ttc     3008 
Cys Pro Pro Gly Arg Ala Gly Pro Arg Cys Gln Glu Val Ile Gly Phe 
        855                 860                 865 

ggg aga tcc tgc tgg tcc cgg ggc act ccg ttc cca cac gga agc tcc     3056 
Gly Arg Ser Cys Trp Ser Arg Gly Thr Pro Phe Pro His Gly Ser Ser 
    870                 875                 880 

tgg gtg gaa gac tgc aac agc tgc cgc tgc ctg gat ggc cgc cgt gac     3104 
Trp Val Glu Asp Cys Asn Ser Cys Arg Cys Leu Asp Gly Arg Arg Asp 
885                 890                 895                 900 

tgc agc aag gtg tgg tgc gga tgg aag cct tgt ctg ctg gcc ggc cag     3152 
Cys Ser Lys Val Trp Cys Gly Trp Lys Pro Cys Leu Leu Ala Gly Gln 
                905                 910                 915 

ccc gag gcc ctg agc gcc cag tgc cca ctg ggg caa agg tgc ctg gag     3200 
Pro Glu Ala Leu Ser Ala Gln Cys Pro Leu Gly Gln Arg Cys Leu Glu 
            920                 925                 930 

aag gcc cca ggc cag tgt ctg cga cca ccc tgt gag gcc tgg ggg gag     3248 
Lys Ala Pro Gly Gln Cys Leu Arg Pro Pro Cys Glu Ala Trp Gly Glu 
        935                 940                 945 

tgc ggc gca gaa gag cca ccg agc acc ccc tgc ctg cca cgc tcc ggc     3296 
Cys Gly Ala Glu Glu Pro Pro Ser Thr Pro Cys Leu Pro Arg Ser Gly 
    950                 955                 960 

cac ctg gac aat aac tgt gcc cgc ctc acc ttg cat ttc aac cgt gac     3344 
His Leu Asp Asn Asn Cys Ala Arg Leu Thr Leu His Phe Asn Arg Asp 
965                 970                 975                 980 

cac gtg ccc cag ggc acc acg gtg ggc gcc att tgc tcc ggg atc cgc     3392 
His Val Pro Gln Gly Thr Thr Val Gly Ala Ile Cys Ser Gly Ile Arg 
                 985                 990                 995 

tcc ctg cca gcc aca agg gct gtg gca cgg gac cgc ctg ctg gtg ttg     3440 
Ser Leu Pro Ala Thr Arg Ala Val Ala Arg Asp Arg Leu Leu Val Leu 
            1000                1005                1010 

ctt tgc gac cgg gcg tcc tcg ggg gcc agt gcc gtg gag gtg gcc gtg     3488 
Leu Cys Asp Arg Ala Ser Ser Gly Ala Ser Ala Val Glu Val Ala Val 
        1015                1020                1025 

tcc ttc agc cct gcc agg gac ctg cct gac agc agc ctg atc cag ggc     3536 
Ser Phe Ser Pro Ala Arg Asp Leu Pro Asp Ser Ser Leu Ile Gln Gly 
    1030                1035                1040 

gcg gcc cac gcc atc gtg gcc gcc atc acc cag cgg ggg aac agc tca     3584 
Ala Ala His Ala Ile Val Ala Ala Ile Thr Gln Arg Gly Asn Ser Ser 
1045                1050                1055                1060 

ctg ctc ctg gct gtc acc gag gtc aag gtg gag acg gtt gtt acg ggc     3632 
Leu Leu Leu Ala Val Thr Glu Val Lys Val Glu Thr Val Val Thr Gly 
                1065                1070                1075 

ggc tct tcc aca ggt ctg ctg gtg cct gtg ctg tgt ggt gcc ttc agc     3680 
Gly Ser Ser Thr Gly Leu Leu Val Pro Val Leu Cys Gly Ala Phe Ser 
            1080                1085                1090 

gtg ctg tgg ctg gcg tgc gtg gtc ctg tgc gtg tgg tgg aca cgc aag     3728 
Val Leu Trp Leu Ala Cys Val Val Leu Cys Val Trp Trp Thr Arg Lys 
        1095                1100                1105 

cgc agg aaa gag cgg gag agg agc cgg ctg ccg cgg gag gag agc gcc     3776 
Arg Arg Lys Glu Arg Glu Arg Ser Arg Leu Pro Arg Glu Glu Ser Ala 
    1110                1115                1120 

aac aac cag tgg gcc ccg ctc aac ccc atc cgc aac ccc atc gag cgg     3824 
Asn Asn Gln Trp Ala Pro Leu Asn Pro Ile Arg Asn Pro Ile Glu Arg 
1125                1130                1135                1140 

ccg ggg ggc cac aag gac gtg ctc tac cag tgc aag aac ttc acg ccg     3872 
Pro Gly Gly His Lys Asp Val Leu Tyr Gln Cys Lys Asn Phe Thr Pro 
                1145                1150                1155 

ccg ccg cgc agg gcg gac gag gcg ctg ccc ggg ccg gcc ggc cac gcg     3920 
Pro Pro Arg Arg Ala Asp Glu Ala Leu Pro Gly Pro Ala Gly His Ala 
            1160                1165                1170 

gcc gtc agg gag gat gag gag gac gag gat ctg ggc cgc ggt gag gag     3968 
Ala Val Arg Glu Asp Glu Glu Asp Glu Asp Leu Gly Arg Gly Glu Glu 
        1175                1180                1185 

gac tcc ctg gag gcg gag aag ttc ctc tca cac aaa ttc acc aaa gat     4016 
Asp Ser Leu Glu Ala Glu Lys Phe Leu Ser His Lys Phe Thr Lys Asp 
    1190                1195                1200 

cct ggc cgc tcg ccg ggg agg ccg gcc cac tgg gcc tca ggc ccc aaa     4064 
Pro Gly Arg Ser Pro Gly Arg Pro Ala His Trp Ala Ser Gly Pro Lys 
1205                1210                1215                1220 

gtg gac aac cgc gcg gtc agg agc atc aat gag gcc cgc tac gcc ggc     4112 
Val Asp Asn Arg Ala Val Arg Ser Ile Asn Glu Ala Arg Tyr Ala Gly 
                1225                1230                1235 

aag gag tag gggcggctgc cagctgggcc gggacccagg gccctcggtg             4161 
Lys Glu  * 

ggagccatgc cgtctgccgg acccggaggc cgaggccatg tgcatagttt ctttattttg   4221 

tgtaaaaaaa ccaccaaaaa caaaaaccaa atgtttattt tctacgtttc tttaaccttg   4281 

tataaattat tcagtaactg tcaggctgaa aacaatggag tattctcgga tagttgctat   4341 

ttttgtaaag tttccgtgcg tggcactcgc tgtatgaaag gagagagcaa agggtgtctg   4401 

cgtcgtcacc aaatcgtagc gtttgttacc agaggttgtg cactgtttac agaatcttcc   4461 

ttttattcct cactcgggtt tctctgtggc tccaggccaa agtgccggtg agacccatgg   4521 

ctgtgttggt gtggcccatg gctgttggtg ggacccgtgg ctgatggtgt ggcctgtggc   4581 

tgtcggtggg actcgtggct gtcaatggga cctgtggctg tcggtgggac ctacggtggt   4641 

cggtgggacc ctggttattg atgtggccct ggctgccggc acggcccgtg gctgttgacg   4701 

cacctgtggt tgttagtggg gcctgaggtc atcggcgtgg cccaaggccg gcaggtcaac   4761 

ctcgcgcttg ctggccagtc caccctgcct gccgtctgtg cttcctcctg cccagaacgc   4821 

ccgctccagc gatctctcca ctgtgctttc agaagtgccc ttcctgctgc gaagttctcc   4881 

catcctggga cggcggcagt attgaagctc gtgacaagtg ccttcacaca gaaccctcgg   4941 

aactgtccac gcgttccgtg ggaacaaggg gtt                                4974 

 
           
             381  
             28000  
             DNA  
             Homo sapiens  
           
            381 

aggtgacccc tagctctgga aaggaccgtg ctcactggag gagaggaagg tgccattggt     60 

tttgaccctg tggaggagct gcgaggtcac ccagggagag ggcaaggagg tgaccgcaga    120 

ggatggggtg tggaagcctg gtgaccaggg cagcagtggg aggcctctct cggggtagcc    180 

ttcagggaca ggcactgccg acttttgttc cccatttccc gcctctcgcc ccccaagccc    240 

agacctgagt ttggggggcg agaggcggga aacggggaat gtggcctgag catttcctga    300 

gggcatggcc tggctacctc gacgccagcg ccgagctgag cagtctgcac cctggagcat    360 

ttgttgactg gctgcttgac cagcgcgcct cgcagagggg aaggcagggg cgtcggaggg    420 

gcgcagcgcc ccctgcagcc ggcgtggagg cggtaggagc ggcgcggaga aggggagatt    480 

ctcggaggag gtggggggcg cgcagtaggg gctgggcccg gctctggccc cagggccgcg    540 

ccaccccgcg tgggggccga gccctgatca gagtaggagg cggcatctcc tctgggactg    600 

cgaggagcgc ggcggtggcg cactgatggg aggggaccac acggcaacct cggggcgccc    660 

cacccccggt ttctgacacc cggcaggagc ccaggcggag gaggggaggc agctttgcgg    720 

cgccggcgca cgcctcgccg actcacgcgg aggtgtgagc ggggcccccg cggcccgcgc    780 

tgaccccgag gccccgtgcc cccgccgccc gggcgccctg gggggcgcgc gccgggccgg    840 

ggcgctggca ggcgacgccc tccaccgcct ttaaagcctg gggcgccccc ggaccccccc    900 

ccggccccac cccgcggcgc ggccccgccc cctcatgcat atgcaggtgc gcgggtgacg    960 

aatgggcgag cgagctgtca gtctcgttcc gaacttgttg gctgcggtgc cgggagcgcg   1020 

ggcgcgcaga gccgaggccg ggacccgctg ccttcaccgc cgccgccgtc gccgccgggt   1080 

gggagccggg ccgggcagcc ggagcgcggc cgccagcgag ccggagctgc cgccgcccct   1140 

gcacgcccgc cgcccaggcc cgcgcgccgc ggcgctgcgc tcgaccccgc ccgcgccgcc   1200 

gccgccgccg cctctgccgc tgccgctgcc tctgcgggcg ctcggagggc gggcgggcgc   1260 

tgggaggccg gcgcggcggc tgggagccgg gcgcgggcgg cggcggcggg gccgggcggg   1320 

cgggtcgcgg gggcaatgcg ggcgcagggc cgggggcgcc ttccccggcg gctgctgctg   1380 

ctgctggcgc tctgggtgca ggtgagcggg gcggcggggg cggcgggggt cgcggacggg   1440 

gcacaccggg ccgcccctag gggccgggcg ggcactgcct ggggccgccg tggttcggaa   1500 

gccctcgagg ctgcgcgcgg cggctggggc tccgggcggg cgcggctggg tgggggcggg   1560 

gcggcggggc ctgttccccc acccctggcg cccggcccgc cgaccccggc ccgcgcctcc   1620 

ctccgctctc ccgctgcctt atttttaggc ggcgcggccc atgggctatt tcgagctgca   1680 

gctgagcgcg ctgcggaacg tgaacgggga gctgctgagc ggcgcctgct gtgacggcga   1740 

cggccggaca acgcgcgcgg ggggctgcgg ccacgacgag tgcgacacgt acgtgcgcgt   1800 

gtgccttaag gagtaccagg ccaaggtgac gcccacgggg ccctgcagct acggccacgg   1860 

cgccacgccc gtgctgggcg gcaactcctt ctacctgccg ccggcgggcg ctgcggggga   1920 

ccgagcgcgg gcgcgggccc gggccggcgg cgaccaggac ccgggcctcg tcgtcatccc   1980 

cttccagttc gcctggccgg tacgtgcgct ccatccctcg tgctccagcc cttccctctc   2040 

tctccgcgcc ccggccccgc gcgcttcgcg acccccaaca cctgcggccg ggtctgcgtg   2100 

cgagccgcgc gcgcccaggc ggggcggggc cggcaggggg cgcgtgctct ggggacttgg   2160 

tccgcgcctg gccacgtggg cgcgccgggg ccccggggcc accgggagcg gggtcgcggc   2220 

gggggcgggg cggcggcgtc ccgcgtgcgc ggcggtgtgc ggcgtgtgcc tgcgtcgccc   2280 

tgcgcgtgtc tgtctgggtg gggaggcgag gcgaggcgcc ccggtcccgg gcaggccgcg   2340 

gtggcatgtg cgcagcgcgt gctggggctg gtctagggca ggccctgact gagccgcccc   2400 

gggcccgtgg ccagcctgcg cctgccctgc agtttcctgg atgcctgggg ggcacgggcg   2460 

ggcgccgtgg gacctaggcc cgggagagcc taacgcctaa cgcttatgtc ggcagaagcc   2520 

cccgatggtg acccaagatc gttcagagac agagatagtg gatcctggtg cagtgacctt   2580 

ctgtggcact gccctgtttg tgggtttttt tggttttgtt attctggagg ggcagaagct   2640 

gagtcggggc tgtctggtct cccctggcag gtggccagtc aggcaggagc cctggcctgg   2700 

gcgtgctggg aggaggggtg gtaggggtcc agtgtcactg ggaaacaggt actcatccca   2760 

gtgggctggc aggtgggtag tggtaggtgg gcaggcccag gcctcgggcg ccttacctca   2820 

ttgcctggag cacggccttg ccctggtgcc cagaggtcct tccctgcttg gtcattgtgc   2880 

tgggggcctg gaactgggtg agtgcgggaa tgagagcacc atgcagacct gtgatcaggg   2940 

agtagatgga tctgggagcc aggaagtggc tccagtcagc aggaggcacc ggagtgtgcc   3000 

cacctggtat cctgggccct gaagtgattg tgagttgagg gcaatccctg ccgagctcac   3060 

gccagttggg cctgccgtgt gtggctccca gtcctgtgct gtacctttgc agccctggct   3120 

ggcagccttg cctgctgccc ccatcctcac cgcttcctga gctcccaccc gtggaagctg   3180 

gccacagtct cctctggcca tgtcctcaac ccgtgagcac cccgccgagt atcccttgac   3240 

caggggggcc ccagagaggg gaaagtgtcc cccagatgga aaaggcaggg gcgggcatgg   3300 

gagggcccag gcagttgtga gaagcccagc ccctcgcccc cacggcggtg cagcaggcag   3360 

gtctgagcag ggcccgcagc ctgtcatctg cacctgggcc tgagccagcg tggccccaca   3420 

tcgctacctg aggatgtgtt ttctgctcga gttggcagca gtgggtgtgg gggcagggag   3480 

gtcttggagg aatgtggcgg gctatcgcgt gtccgccctg gctcttcgcc ccgcgggcca   3540 

gccggtcagg tgtgggatgg gaccgggtag gcccttgcct tccttggagt ccgggcactg   3600 

ggtttcgggg ccagctcacc tccctgcctc ttgcttccag ccggttcctc gaatgcccca   3660 

ggagggggca ggcggcctgt ctctgggttg ggggccaggg cagagtcata gctgcgtgtt   3720 

tgggggcagc cctggtctcc tgccatgtgg cctggctgcc gggcgggagc tgtgccgtga   3780 

tgccagcacc ctggtatttg cactcgggcg gcggcagtcc ctggccatgc tgccctggct   3840 

tgctgaggtc cagctctgtg cggtgagctg aggtgtactt ggctgtgatg ggaaggcaag   3900 

gaccgagttc aggctccctg ggacctgagg aggggtttca gcctggaggc tagggtggca   3960 

tcctgcccag gcccgtgggg cttttgggct ccttggagta aagggaatga gagggccttg   4020 

tggaagagga gtgggggagt ctgggctctg cattcgctcc ctccaccccc tgccccctga   4080 

gtgactctcc caccttgtgg tctctgctgt tgacccaggc ctggctgggt ggccctctgc   4140 

cccctggcct ggcttcttgt ggccggggtc tgtgtgctat tagtcatgga tctgtgctgg   4200 

tctcgggctc agcttccctc agtgggtggg cccagggtct tgaatgtgga gaggtggtgg   4260 

accacatgcc agcaggctgc ctggctgccc ctcctctcct ggctccaccc ccagacgtcc   4320 

ccaggaggcc ggtgtcagcc tgggttggtt ctggtgcctg gcttgtagct ggcagggtga   4380 

ggccacattc tcccagctgc gtgtgtgcac gcaccccggg tgctctgtag gcatggcagg   4440 

tggtgatgga ggtttgggga ggagtagtgt catgctgggg gcagcaggga gctttgctct   4500 

ggggcctggt aggtggcagg cccaggggac acctgctggc tgagggagga gcagggtggt   4560 

ggcagttggc cgtgacctgg gcagccaggg ccccaccctc agaggtgcag ctggaagtcg   4620 

tgcctgcctg gctggcccat ctctggagcc aggagcccag gagcctgcct gccagcgagg   4680 

gtctttcttg ttctgtcttg gcatgtgtgt gctgggctcc agggcagctg tgcggggtgg   4740 

tgtggctgga gcatggtccc cgtgacagat ctgggcttat gaggagaacg ggatgggtga   4800 

aggccctgta gatacaggag gtgggcctgg ggctgaccct gcgtctatca gctcaggagg   4860 

cctgaggtcc tgggccatca gaagggctga gcttttctca cctgtgaaag gggcacactg   4920 

ccgctttttc attgcaggtc tcacgaagtc agatggggct gctggactcc cagctcgggc   4980 

tctgcttgtg cccccagccc ggctcccaga cctgtccagt tcctcccctt cccagccttc   5040 

cctaccctct cctttgcccc ctagggagga aggtttttac agagcccacc ccctgcatcc   5100 

agccgcccta gggctcaagg tgggccaggc tgaggtctgt gcctggagct acctaagctg   5160 

ctcgtggcag gtgtgaggtt cagcaccact ctgcttcctg ttttttctga gcttgggctg   5220 

gggatgacag ggccctggcc tccccaccct accttcaggg gatcctgtct gcacactggg   5280 

gaccaccccc ctccttccca caccttccca gtagggacca ggagagctgg ctggtctggt   5340 

atggaatgtg ggcatctggg ttcctgtgtt gggtgggcat cggtctgttc ctcctgccat   5400 

ggccctgggg cccagagccc tggggagaac tcagggcatg tccgccttgt acattggggg   5460 

tctggttcaa agctttggta tgggggcagg gtggggcatt cagtgcccag gcaacacggg   5520 

gaccattgga gccagggagg actgcccttg gccagggagg attggagagt gggctggggg   5580 

tttgtcgctg gtccctgagg gtgggctgaa gggtcaaagc cgcagcacga taggaaggct   5640 

gggaggtgga ggggcgggtt ggggagcagg cggcaggcct gggtgggagg ggactgctgc   5700 

tctcaggggc cctcctgggc tgctccatgg tgtctttatg aggggagcaa gctaggccag   5760 

tgaaggggtg cttgtggagc caggcttcgg cctgagctgc tgctggtggt ggagtggggg   5820 

caggaagaca aggatctgca atcccaggcc ccagccacag tcgctatccc cagaccccag   5880 

gcctgagcgg ggtccctgtc cccagaccct aggcctgatt ggagtccctg tccctagacc   5940 

ccaggcctga gtggggcccc tatcctcagg ccccaggcct gagctgggtc cctgtcccca   6000 

ggtcccagga ctgagcaggg tccctgtctg cagacctgag gcctaagcaa ggtccctgtc   6060 

cccagacccc agatctgagt gaggtcactg tccccagacc acaggcctga gcggagtccc   6120 

tgtcctcaag accacaggcc tgagcggagg ccctgtcccc agaccacagg cctgagcgga   6180 

gtccctgtcc ccagaccaca ggcctgagcg ggattcctat ccccacatcc caggcctgag   6240 

cagggtgtgg ctggcatcag ttgtaccctg ggctttgtgg caggtgctag ccggccctgg   6300 

ctgccaccgt cttcacggtg ggggacctgg gacctagagg gggtgtgctg gggagtgggg   6360 

gtacacccag gcaaggccct ggctggtctc tggtgtggag catgggtgtg tgtgttcctg   6420 

cgtgggatgg gctttggtct gctcctcctg ctgcggccct ggggcccaga gccctgggga   6480 

tggtgtttgc ccccacccct tcttccctgc ctcgggtgac aatggtggca gaggcctggg   6540 

cctctcagaa gctcaggttt caggaaatgt atctgtgctt ggagctcctg gcgcctgcac   6600 

caagcgctgt gctccgtagg gggcgggagg ctgatgcggg aggccgagga gaagaaacca   6660 

agtcggggcg ttggtggggc agcaggtcta ggaggctgtg ttgtgttggc ctggaccgtg   6720 

cagggccctg gacctggggg ggccgttagc ggggcagcag ggaggctgtg ttggccttga   6780 

ccgtgcaggg ccctagactt gggttgcctg agttttggga tgctgtagat tggggtacag   6840 

tgggcagtgg ggtgccgtgg acttagggtg cttggcattt ggagtaccct gggccatgag   6900 

gtgtgctggg ccatgcagtg ccctgggctg ggggtgccct ggacctggag tgccctgagc   6960 

tttggggtgc actgggccat ggggtgcaca aggctgtggg gtgtactgga tctagggtgc   7020 

cctgggcagg agggtactct gtactttggg tgccttggac ctgaggtgtc ctgggcttta   7080 

aggtgccctg gaccttgggg tgtgctgggc catgcagtgc tttggggctc tggggtaccc   7140 

tgggctttgg ggtgccctgg aactggggtg ccctgggtct tggagtatgc tgggccatgc   7200 

agtgccctgg gctgtgggat actctgggct ttggggtgcc ctggacctgg ggtaccctgg   7260 

gctttgaggt gccctggcct ggggtggaac atctattgtc ttgtctgcct gtcctctggc   7320 

ttgtgccact gctgttgccc ctgcctgggg acaggaggag gggtttagac ttagccttga   7380 

gggttcgggc tggggaggag gcaatcagat ggtgggagat gaagttgggc tgcgggtctg   7440 

cttgtgcggt gggggtgggt caggccgggc ttgtagggag aggcttagct gggcctgcag   7500 

gggtgaagcc cttccccctt ggcctccaga gactgggcag gggcatagcc ctgctaggct   7560 

ggccttgagg gagggcctgg gttcctctcc ctgcttgccg gggaacctgg gcaggtgatg   7620 

ggtctctcac ctgtccccag acccccagcc cacacatcgc ctattgcccc tgccagcgcc   7680 

aggcccacat ccccacatgt cccagccccg ttcctagaag ggcaacatgc ccgccaaccc   7740 

ccgcccaatc caggccctat agtccctcct gtgttctagg ggtttggtgt tgacaaaacc   7800 

ctgtcccaga tcgtggcccg ccaggcagga aggacagggg tgagaggttg ctattcgcag   7860 

aggaggcaac tgagtcctgg aaggacaggg gtgagaggtt gctattcgca gaggaggcaa   7920 

ctgagtcctg gaaggacagg gatgagaggt tgctattcgc agaggaggca actgagtcct   7980 

ggaaggacgg gggtgagagg ttgttattca cagaagaggc aaataagtcc tggaggctgg   8040 

cccctaggga agaaggggag ctgggagagc tggcaggtgg ggtgaggcag gtaccgcccc   8100 

gtcagccagc tcaggttcac tctggatgac ttcctgccat ccaggtgtag ggaccccagc   8160 

tggcgggcgg tgaggccctc tcggcgggcg ggcaggcaca cgtccctgcg ggagcaggta   8220 

accggagccc tgggctcagg cgaaggtggc agtaatctta cctgagtggc tggcatgagg   8280 

tttcctggga gtcgagagga actccctgct ggccctgaag cccaggtgtg gctgtgccgg   8340 

gagaccgggt ggcctggctt ttctctgcct gccccgtggc cagagctgct ctcagaccca   8400 

tgctggcccc atcctctgac ctcactattg ctgcttcctg gtcctgctgg ttcctgtcca   8460 

gcggctacag tgactgttaa agcctggtgg gtcccagtcc tcactcagac ccccaacaac   8520 

agacctcact cagaccccca acaacagacc tcactcagac ccccaacaac agacctcact   8580 

cagaccccca acagacctca ctcagacccc caacagacct cactcggacc cccaacaccc   8640 

gaacagacct cactcagacc cgcaacagtc acccacttcg cttagcctca ggaaggaagt   8700 

ccgtggtggg gtctggatct gtggtatgac cccactgtcc ccgtgggcta tgcgttctca   8760 

gcccctgggc cttcttgtgg gctctgccat gcagctcctt cacttcctca tgccctgcag   8820 

cctcaatctc aatgccacct gttcaaagcc tggcctggcc tttttttttt ttttttgaga   8880 

tggagttttg ccctcgttgc ccaggctgga gtgcaatggt gcgatctcgg ctcgctgcaa   8940 

cctcggcctc ctgggttcag gtgattctcc tgcctcagcc tccagagtag ctgggactac   9000 

agacacctgc caccatggct ggctaatttt tatattttta gtagagatga ggattaaccg   9060 

tgttgaccag actagtctcg aactcctgac ctcaggtgat ctgcctgcct cagcctctta   9120 

aagtgttggg attacaggcg tgagccactg tgacccgttg gcctggcctt attggaacaa   9180 

cagcccctgc cccctgttgc tttccccgag ccccgctggc tataggttgc cgtccttggt   9240 

ggcagaggca tgcctgctgt acacttgatg tgaacgaagg aaggaaggaa cgaaggaagg   9300 

agccaaatgc cagacgcctg ggaagcggct gggtgctcca ggtgttaccg ggggtgggga   9360 

agggcttggc caggtgcagc tgcgagggtg gtgctccagg cagatgggtt gataggctgg   9420 

ggtgggtggg tgggggtggg caggagcctt gggaacccca agggtgctct gagctgagag   9480 

ggcgtggaca gagtcctggt gggggtgtgg atggagccct ggggggtgtg gatggagctg   9540 

acgggggtgg tttgtggaca gagcccttgg ggggtgtgga cacagtcctg ggggtggtgt   9600 

ggacagagtc ctggggggtg tggatggagc cctggggggg tatggatgga gcatgttggg   9660 

gggtgtggat ggagctctgg gggggtatgg atggagccct gggggggtgt ggatggagca   9720 

tgttgggggg ggtggacaga gctctggggg gggtgtggac ggagccctgt tggggggtgt   9780 

ggatggagca tgttggggtg tgtggatgga gcatgttggg gggtgtggat ggaactcggg   9840 

gggtgtggat ggagctctgg ggggtatgga tggagccctg gggggtgtgg atggagcatg   9900 

ttggggggtg tggacagagc tctggggggg ggtgtggacg gagcatgttg gggtgtgtgg   9960 

atggaactct gggggggtgt ggatggagcc ctgggggggt gtggatggag ctctgggggg  10020 

gtgtggatgg agcatgttgc ggggtgtgga tggagccctg gggaggtgat ggagcctgtt  10080 

ggggggtgtg gatggaaccc cgttggggga tgtggattga gttctttggg ggtgtggatg  10140 

gagctctggg gggtgtaaac agagctcggc ggggggtgtg gacggagcct tgggaggcat  10200 

gtggatggaa ctctggggat tgtctggcgc ctgtaggcag aggtttgcgg gccctggtga  10260 

cctcagggag ccctggagat gggcggggac tgggccccgt ggcctggcgg ggccatggcg  10320 

gatgtgggaa aacgggttta aggggagctt aagaggtggg attgagggtc tgttgtcagc  10380 

tcgacgtggc tagggagggt tctaggagcg ggttggggat ggccccccac ttccatcctg  10440 

tgctcctacc tgggtgagcc tcctcgggcc gtccccgggt gttctgcagg caggggctcg  10500 

ggggcggggc cggggttgcc cagctgtgag tgaggcccag ggtcagcagt catgttgggc  10560 

cctagttgtc tgtatttgag ggacagtcgg aggtgtgggg cgggggactg ggtgggggtc  10620 

ctggaggctg ggctgggtgt ggctggtagc acgttggtta ggggaggggc tggacgtggg  10680 

agtgcagtct tctgagacat cttgggaggc caggcctgtc cttagctgga tgaggccgag  10740 

gcactgggac gtgcgtgggg tgggcggcgg gtgaggacca gggaagggct ggcaggcgtg  10800 

gggttgggcc ttgctgggga agtgtggttt tccccagctt agccaggcct tggggctggt  10860 

tggatggggt gtgctgaggg atggagtgag cctggcctgc ctggacactg cccaacgcag  10920 

catccccccg gggtgggaag ccagcaggcc ctgaggtgac tcagccccag ccccctcctc  10980 

tgggccccac ctggaaggag ccagggctgg gctcaggggt caagagcaca ccaggggtag  11040 

actggggggt tcctgggcag tgagggctga gaggctgtgg aatgtgggta cccagtgctg  11100 

ggtagtacag ggcatgtccc gggggtccca cctgtctgag catgtctgtg agtgacggtc  11160 

tccgtgggct gcactaggcg gagcaggggg ccagccctgt ggtctctttg cttggctgac  11220 

agcatcgcct gtcgccatgg ctggggtaca agggccaggt ggcccggggg cagagggggc  11280 

atagtggcca tggtctgagg ctgtgctggg cagtcccagg acctcttggc ctcagtttcc  11340 

ccaactgtac cgcaagggcc cctcctgcca cctgttctgt gtgagggtgg aggtaggtgt  11400 

gggtttgcct gtgtgctgta tgcctgcagg acctgagctc cggcctgttg gggcctctgg  11460 

ctgggcgccc tgtacttggc caccccgtgc acttggtgga ggccgccagc gtggtgatgg  11520 

ggccccacgt tctcccccgt ggtcaccccc agtgaggcac caaggggcgt tccacaggaa  11580 

acgctcgggt cccggctgcc catggggccc ctgtctgtgg ccactccagc caggctgccc  11640 

tttgcccacc tctccccccg gtcgctcttc ctgtgctccg tgctgacttg agccagctca  11700 

gggcaggctg ggcctctggc accccaacgg tagggagccc aggcccctga gcccgcgtgg  11760 

cctggagggg cagtctccct cccttgagct gggtcatttt tgggtctgca gaggatgtgg  11820 

cctgaggatg aggagggtgg tgggtccctg gctggggagg aagggccaga gcctggcaga  11880 

cccaggggca gcgtctgagc cctgggcctt gtcccaccct gaacgaggca ggcaggtgtg  11940 

gcctcaggta cctgacccgc ctccccatgt ctgcagcgct cctttaccct catcgtggag  12000 

gcctgggact gggacaacga taccaccccg aatggtgagt gagccctggg ccaggtggca  12060 

gctcctctca gcttcagcgt gcctgtggca gggcccagct cctctgtctg cttgggacaa  12120 

agccttgctt taccctgagg atcatgtgtg ctgtttccct ttttgctttg gctgccagga  12180 

agctctgcca cgtttgggac ttgcagagct gtgcatgcac tctcttcccc agtcctggct  12240 

ttgcctatgt tgttctcctc ttgggtgtgc tcttttgggg cccatggcag tgacttagtg  12300 

gaggggacac ccttgagtgt gtctctggct ttgtggcccc ctctgcttgt ctgtactgga  12360 

gcatggagcc ttggtggccc tctccctgag gcaggggctc tgcagggccc tgcaggggta  12420 

acgggatgac ttccatgggt gaatgcagaa gcacccacag gccaagggag cagctcgtgt  12480 

gaaggtctgg gcaggagcgg gctggctgtg cagggggagc agccggggct gggctcagat  12540 

catggaggct ggcaagccac tgagaggaca cgggctccgc ctggcaagct gtggctgcct  12600 

tatggagggt gggctgtggg gccaggacac agaccgagga ggagctgcca cgtgaatctg  12660 

ggcgtgtcag ggtgacttgg accaggggca gtctgggggt gagaggggct ctcagaagtg  12720 

gaggcatggg gttggccaat gggttggagg agggagagcg gggccagggc atcctggctg  12780 

ccagcagagt ggaggggctg ttttcagggc agggacggcg gtgggggtgc ccaggtgggg  12840 

agcagcagtt gtggggaccc cagcggctca gggcaggggt gtttcctgag ggggtggcag  12900 

agagacaggt gggctgagtc ccaagcaagg tcgtcagggc tcttgacaac gtgagcctgg  12960 

agaggctggg gcggccggga cgccccttgg ggagtgggcc agcacagtgt cctcccaggc  13020 

cttggcccga ggcgggagag gtggggtctg gaggacccgt tcacctttta ttgtgcaaaa  13080 

cgtcgagcct gtgcctaagc gcagggaccg gcatcacgga ctttgcatac cagcgccagc  13140 

agctgtggtg cccctggccc ctggtctcct ggtggcttac ttaaagtgag gcttagacag  13200 

cgggtcacgg gacctatgcc tgtcttgggg gcctgagggg aggcttgtct taaggtgggg  13260 

acggtagtgg tgtttggcac ttctgggagc aagtcacagc gcaggagagg ggagggcaac  13320 

tgagcaccat gtccgtgctg tcgagggctg gacacggcgc aggtgggtgc aggtgttgga  13380 

gcagggctgc aggtgggtgg gcacaggtgt gggacgtgag actcacgccc tggcagcagc  13440 

cgtgccttct ctgtggagcc tgtggtctca gcagccctcc ctgcagggcc cctggcccct  13500 

agccgggccc cccgaccctc tgcgtttagg gtgggagcgg ggcgcaggct tggtggcggg  13560 

agggagaggc ctctcggggc cctgagcttt ctgtagcagc ctggccgggg gccctgccct  13620 

ccgtgtgctg ctgcctgctg tgccccggcc ttgcagcagc cgcaggcttc tgccccgtcc  13680 

ccgttgttcc tggaggaccc ctggccgggc tggtttctct ggcctgtgct gactctgccg  13740 

cctccccaag aggagctgct gatcgagcga gtgtcgcatg ccggcatgat caacccggag  13800 

gaccgctgga agagcctgca cttcagcggc cacgtggcgc acctggagct gcagatccgc  13860 

gtgcgctgcg acgagaacta ctacagcgcc acttgcaaca agttctgccg gccccgcaac  13920 

gactttttcg gccactacac ctgcgaccag tacggcaaca aggcctgcat ggacggctgg  13980 

atgggcaagg agtgcaagga aggtgagggg gccgctgggc cgcgtggagg gcagggaggg  14040 

cctcgggcag ggccccgggc acaggccttg cggccaggct ggctgcagct gtgcctctcg  14100 

ctcctctctg ttcgcagctg tgtgtaaaca agggtgtaat ttgctccacg ggggatgcac  14160 

cgtgcctggg gagtgcaggt gagtgtgccg ccggcccgtc tttgccctcc caacctttgc  14220 

cctcacgtcc tcactggcac acacagcctt gctgtcagga gtcgcccgga gctggctgga  14280 

ggttgggcac acagctgtga gagccgggcc ctgagctcgg gaggctcctt agtgcagtag  14340 

gtgcgtgtct gagcatggga tgtgtctgat ggcggcagcc atgtgaggac agtgaggaga  14400 

gactggggag gctggctgga cagtcacgtc accgaggggc agagacccgg aggctgcaag  14460 

ccacccagag atggggcatg ggcagaggac acggtaaccc tgcccatggg gagggggtgg  14520 

gcggcgagcg gccggcaagt gacaccagca ggcgaggggc ggcagagcag accagtggtg  14580 

ggagctgagg cctgcaggac cagggacaga ggaaggggct gctggcaggt ttgtagctgg  14640 

gcaaggtggg tggaagggct gtggtagctg ttgagtgggg aagcaccaga cgggaggctg  14700 

tgagggggag gccgctgtgg ggcatgtggg ggtggtgggg gaggggccag cgggatgggg  14760 

agggggtcag tggaaggggg agaggcgcac gggtcctgca gacatcctgg ggtggagccc  14820 

aggggtttgt ggatggattt gatcaagcag gaagggtgtg gagtcaggga gaaccccaag  14880 

ctgctgagta gcagagccat ggtggcagga ggaatgccac agaggagcag gcggggccgg  14940 

ggttagctgg atgtggagag gcgatgcctg ccctgtccct ggagacaccc agaaagctcg  15000 

tgggagaggc ctggcctgcg ccacgcgggg cctgtggggg gtggcattca ggcggtgacg  15060 

ggaaagtggg gaaggcagag aggagggagg ccaaggagca agtcccggct gccacaggtc  15120 

agggcggatg gatgaggagt cagcaagggc ctccacaagg gagtgtccgg ggtcttacag  15180 

ccaagtccag atggtggagg cctctggacc cagaccagag tgtgggggat ccagccaggg  15240 

gggctggcag ctttgcccta gagtggagca gagaagtcag cagggcaacc agaggggctg  15300 

gggcccaggg tctggggtgg gcacgggctt cgagccgtgg cgctcactgt gcgtgagcaa  15360 

gctggggagc ccgagagagg ggcgcgagcg ggtggagaga cagcaggtgg aggtgagcac  15420 

cgccctccag ccagccttgg attgcagggg tcccaggacc tccctctgtg gagtgggttt  15480 

gcctccatgg gacgaggaca ctggggcaca gagagcctac tgatttcccc agggtcacac  15540 

agcgtggcgt tttggagagg agtcggggag tttgggaacc agctgagttg ggagccaagg  15600 

tggggaggtg ggtgaccctt ccacaggccc cacggttgag tggcctggag ggtacagtga  15660 

ggagctttcc cggccagtcc cagagcgggg aggcaagcag ggctggggcc gcccacccgg  15720 

tcacttgcac acacagggat tcccggcagg ttgagcgagt cccaagtcag ctcagaaagt  15780 

gcaacaaggt ggacctggtc tgggcagatg tagatgtaga tctacgggag tcggccccac  15840 

tcaccctcgc ctggcccagt gtgcatcaca caacctggat ggcagtgcca ccctccctgg  15900 

atggctgctg gctggcagct tgaatgtcac accaaggctg gaggaaggca gcagagaagt  15960 

tggccatccc tgccctttac ccgcaggaag atgagccgga gtctgggggg cctggtgggt  16020 

gggggcagta ggtgagctcc gcctgcccct cttgctggcc ctgtcgggga ggcccagctg  16080 

ttgctgacag cctcggctca ggttccagtg caggacgccc ccccaccgga tgctgcggag  16140 

atggccatgc cttcctgccg ccgcctctcc agggccctgg ggctgctggc tggggaaacc  16200 

aggaggtggg ggcctggtgt gggctgccct gcccagggtc gagagcacgc ccttgggacc  16260 

cacgaggtct gggctctgag cccggctgtg gccgctctct ggccgatgac ccaaggtgtg  16320 

tcacagcccc gccctgagcc tgggtctctg tgtctgtgga ggagggattc taggcgggat  16380 

gtgaggccac ccacgcggac cactgtgcat gctgggctgg atactggaga cacgttcttc  16440 

ccggcctcag tttccccatt tgtggcagct gaactgggct gataggcctt cggtgctggc  16500 

tgtgtggctt gagggcggct caggaagggc cgtggttctt tccttttaca aaaataaagt  16560 

gtggcgggtg ccggtgtgga agtgacgtgg cctggatgac attcccgtcc tgcaggaccg  16620 

gagagttcta ggaagggccc cccgggagtc ccggcagggc ctggatggca gcctgctgag  16680 

ccttggggtc gttgcaggct ctctcccctg acggaggcac cctcaagtca ggccatgttc  16740 

taccctggcc acctgccctc tcctggggga ctcccaagac aggacgttgg ccgatagcct  16800 

ggggcagggc gagtcctggt ggttgtgtcc tggggggtgc agctgggggt gcagctggag  16860 

ctcctgcaga atcaggaact accctgggca gggctggccc aggccagcct gtgggcctca  16920 

gtagccccat ctgtgagatg ggtaccttgt gggactttac tgggagcgag cgaaatgact  16980 

gcctttgagg tgggggcgag ggcacgtgct gtgcccaggg ccacatggcc gaggcagagc  17040 

caggagtgct cccctgctgc ccgctggcct acccagcccc tggtgcctcc cggccctggc  17100 

agcaccttgt gagtccgagc cggcattctc atccccgggg tcccggcagg gccttccttt  17160 

cctggtgcct gctctcgggg cccagctcac gggtgaatcc caaaatagct cagggaggag  17220 

tgacgggaca gctggggctg accgtcggca gccagcggcc gggaatgccc gtgacagtgg  17280 

ggctggccgg cagggctgca acccctgcct ggctggggct gctccagttc aaaggcctga  17340 

ggccgcccgc cggccctggg tgtggcgtgg gtgactgtgc ctggctcccc tgccaccctt  17400 

tcaggcacca cagctcactg ggtcttgcgc ccctcctcct tcccccaggt gcagctacgg  17460 

ctggcaaggg aggttctgcg atgagtgtgt cccctacccc ggctgcgtgc atggcagttg  17520 

tgtggagccc tggcagtgca actgtgagac caactggggc ggcctgctct gtgacaaagg  17580 

tagtggtagg gggcggcagg cctaatgctc tgccatcgaa gtgtgggttg tgggggagcg  17640 

gggggccggc ttttcccctg agcatcccac ccctgccccc agacctgaac tactgtggca  17700 

gccaccaccc ctgcaccaac ggaggcacgt gcatcaacgc cgagcctgac cagtaccgct  17760 

gcacctgccc tgacggctac tcgggcagga actgtgagaa gggtacgtgg ggggctggcc  17820 

acccaaattc tggccaggca gggactggtt ccctggggag ccggtcaggc cccatccctc  17880 

tggcgtcctg tgtggtgggc ccctgacccc cagcttggga acctgtgggc ttggggagga  17940 

gtgcttgtgg aaagctgggg gcctggctgc cagctctgcc ccctccccgc ggttctacag  18000 

ctgagcacgc ctgcacctcc aacccgtgtg ccaacggggg ctcttgccat gaggtgccgt  18060 

ccggcttcga atgccactgc ccatcgggct ggagcgggcc cacctgtgcc cttggtgagt  18120 

gtctgcacgt gagtagggga ctcctgccta gtatcagtgg gggtctggga gtggggcaac  18180 

tcgctgggga tggggtgcag tggtcaagtc cacacgtgtg gctgcggctg gcttggcgag  18240 

gacaaatggc aggaagaccc aggcttgcag cgccacctgc ccatggggac cttattccca  18300 

cggctcacac tgccagggcc ccacctttct ccaccctctg cagacatcga tgagtgtgct  18360 

tcgaacccgt gtgcggccgg tggcacctgt gtggaccagg tggacggctt tgagtgcatc  18420 

tgccccgagc agtgggtggg ggccacctgc cagctgggta agggctccga gcgagtgcat  18480 

gggaacgtgg gccgcgcatg cgggctgcgg gggctgctgg ggctgcgggg gctgctgggg  18540 

ctgctggggc tgctgggctg cgggtgccag gtgcccgtgc tgcagggggc aggcagggcc  18600 

cgagccccac ggctcccacc ttgtctcttt cacagacgcc aatgagtgtg aagggaagcc  18660 

atgccttaac gctttttctt gcaaaaacct gattggcggc tattactgtg attgcatccc  18720 

gggctggaag ggcatcaact gccatatcag tcagtatggg gggtgggcgc cggcgggtgg  18780 

gccgaggcac atgggacccc gcctctgacc ctgctcctct gcccccagac gtcaacgact  18840 

gtcgcgggca gtgtcagcat gggggcacct gcaaggtgag gcggggccag gagggtgtgt  18900 

ggcgtgggtg ctgcggggcc gtcagggtgc ctgcgggacg ctcacctggc tggcccgccc  18960 

aggacctggt gaacgggtac cagtgtgtgt gcccacgggg cttcggaggc cggcattgcg  19020 

agctggaacg agacgagtgt gccagcagcc cctgccacag cggcggcctc tgcgaggacc  19080 

tggccgacgg cttccactgc cactgccccc agggcttctc cgggcctctc tgtgaggtga  19140 

ggtctgcctg gtcaccctgc cccacctgct gctctgggag ctgtagggca ggcctcgtcc  19200 

cctgaccatg gggcctgagt gacccagggg tgctgcaggg gaagttgtcc ccaaggcgtc  19260 

ccaggctcag ctctccactg ggtgccaggt gggcaggcgg ggctgtcaca ggtcaccagg  19320 

cttggccccc tgtggccatt gcttgttgtg atgggtttcc tggtggcctg ggctaggagc  19380 

ccccgggctg ctggctgccc aggcctatct gtccatctgt gcactccctc gggactggag  19440 

ggcagggggc tctggtgggc agagcacatg gggtagggtg ggtgcctgat ggtggagagg  19500 

tatacacctg tcataggtga gtcctgggtc ggagtgggca tctctctcag ggctgatgct  19560 

ctcgcctccc tctgaccatc tgttggtact ggaccccccc cacccacctc cctaccaccc  19620 

tcggccgccc acgatcctgc cctggccttg gtgcagagga tgggcctcct gtccagaggg  19680 

cttcttgggg cccagggcag gggtctgacc tcaggacctg caagcatggc agtggctggc  19740 

cctggaaaag acccacagtc ttggctctga gggtggccag gcagtgtgtg aggggctcag  19800 

gagctgtcct tcctgccagc agcaggggcc aaggccacac tcctcccgag ggacagtgag  19860 

gaagctgggc tgcagtggag gtgggggtgg gggcccacag gtatctgcgt tcagctaagg  19920 

cctgggcagt ctcaggtggg caggggtctt gggctctggc tggcactgtt aggcccaggg  19980 

cggaggggcc tgggggtccc cagggatcta ccttcgtatg gacagaggcc tggcctgtgt  20040 

tcccggcctg ggcctgggcc taggctctca caggcacccc ccaccctgca ggtggatgtc  20100 

gacctttgtg agccaagccc ctgccggaac ggcgctcgct gctataacct ggagggtgac  20160 

tattactgcg cctgccctga tgactttggt ggcaagaact gctccgtgcc ccgcgagccg  20220 

tgccctggcg gggcctgcag aggtgctggg tgcggcatgg ggtggtgggg gaggtggtgg  20280 

ggcaggggcg ggcctgactc ctgactgtac tgcctgccat agtgatcgat ggctgcgggt  20340 

cagacgcggg gcctgggatg cctggcacag cagcctccgg cgtgtgtggc ccccatggac  20400 

gctgcgtcag ccagccaggg ggcaactttt cctgcatctg tgacagtggc tttactggca  20460 

cctactgcca tgagagtgag tggccacgaa cggcgggctg gtggtggggc tgggctggcc  20520 

tgaggccctg gctcaccccg ctcgcctctg cagacattga cgactgcctg ggccagccct  20580 

gccgcaatgg gggcacatgc atcgatgagg tggacgcctt ccgctgcttc tgccccagcg  20640 

gctgggaggg cgagctctgc gacaccagtg agtgttccag cacccgccca cacggcctgt  20700 

gcctccaccc ctgtgggccc cttatcaccc tgagatggac cgctgtctgg gtgcggcagg  20760 

ccccgtaccc agaaaggcct ggccaggggg tgctgccacc atggggtgga gtcccaggct  20820 

gcccccatgc ccgaggccag ctcccccggc ccgacgctcc tcccccgccc ctctctgtcc  20880 

tcacctggcc cagctccagt gcttcctccc ccgggaagcc ctccctgagc gccggtgacc  20940 

ccccgcccgc tgaccggcgt cctcgccccc agatcccaac gactgccttc ccgatccctg  21000 

ccacagccgc ggccgctgct acgacctggt caatgacttc tactgtgcgt gcgacgacgg  21060 

ctggaagggc aagacctgcc actcacgtga gtgtccgcag gccctggccg cctggggctg  21120 

cccccaggac cctggccctg gcggtctggg gcctgcctgc tgagcggccc atgtgccaac  21180 

aggcgagttc cagtgcgatg cctacacctg cagcaacggt ggcacctgct acgacagcgg  21240 

cgacaccttc cgctgcgcct gcccccccgg ctggaagggc agcacctgcg ccgtcggtga  21300 

ggagcccccg ctgcctctgc gaccgccggg catatgccct cccaggcacc gctccctcgg  21360 

gcgcgatggg ccgaggggtc ttttttgagg gccacacctg ccacctgccc cctgccccct  21420 

gcccccgggt ctgtctgccc tgtctgggtt gggggcgcgg tatggagacc cagggccagc  21480 

ccagggccag gtgagacgct ccctcctcct cctctcctta cagccaagaa cagcagctgc  21540 

ctgcccaacc cctgtgtgaa tggtggcacc tgcgtgggca gcggggcctc cttctcctgc  21600 

atctgccggg acggctggga gggtcgtact tgcactcaca gtgagtgtgg gaggggtgtg  21660 

ggcgggggcc gctttcctcc acccagatga catccctgcc cccgactcgc cccccagtcc  21720 

cttctgccag cccctccccc tgctgcccct gcccccagca aaaggcaccc tccttgatga  21780 

ccctccccag ccccacagcc tgatcacgcc aagccagcct ggacagtgcc tggcacgctt  21840 

ggggggtggg tactgatccc ctgcgttctc ttctcccaaa ccagatacca acgactgcaa  21900 

ccctctgcct tggtgagtgg caccctgggg gccacagcag gggtgggtgg gacttggcat  21960 

accacggggg gccacctgat gcccaccctc tgctctgcag ctacaatggt ggcatctgtg  22020 

ttgacggcgt caactggttc cgctgcgagt gtgcacctgg cttcgcgggg cctgactgcc  22080 

gcatcagtga gtggccagac agccccagcc ctgggagccc ctcagcccag ccgcggtgtc  22140 

aggagtctgg ggacatcaac gtccacgtcc cttgaagggc agtgtggcca caactacttc  22200 

ctgcctctct tctgagcctc agtttcccca catgtctgtg ccctgtgggg ttcctgctgt  22260 

ataccctgcc aagtgattaa gtggggagcc ccagcctggg ggaccagtcc ggggcccagg  22320 

gagctgtggg ggttggagcg tgcagcctga cgtgggctcc tctgtggccg cagggctgtt  22380 

gtccctgggt gttggcccag ctgtctgtcc agcacccctt ggctggtccg acgcagcagc  22440 

tggggctaat ccaggatggg acaggcccac tgcagaagca gacggaggag ggtgctgttg  22500 

ggccagggtc aggctgggct caggaaggcc tcaggcaggc agcagcttgg gctcgggggc  22560 

aggggctgct cctcattgtc ctggggcttg cgcctgtgtg ccactggctc cccgctgccc  22620 

taggccatgc cggtcctgcg gtgggcgttg gcctcactgc actgagcagc ggtggctctc  22680 

cctgcagaca tcgacgagtg ccagtcctcg ccctgtgcct acggggccac gtgtgtggat  22740 

gagatcaacg ggtatcgctg tagctgccca cccggccgag ccggcccccg gtgccaggaa  22800 

ggtaggcccc gtgtgattgc cctgggttgg ggcgggttgg ggggcatggg tgacacccag  22860 

ccccgagggc cagatgccca ctgctgaccc tcgagcccct tctccccaca gtgatcgggt  22920 

tcgggagatc ctgctggtcc cggggcactc cgttcccaca cggaagctcc tgggtggaag  22980 

actgcaacag ctgccgctgc ctggatggcc gccgtgactg cagcaaggtg agggcagccc  23040 

gtgagccgcc ctgccctacc cgaggctggt gcacgctgac cctggccact ctgtgagatc  23100 

aggaggcggg tgctggggtc cggatggact gagagccgtc tgccctcagg gacacccagg  23160 

gaggcgagag ctcagccagg ccccatgctt cgatgtgcag ttgggaaaac aggcctggtc  23220 

tgggtcctgc cttgctccgc ctgccctttc tgatgtcgag cttggcctgc ctccctggga  23280 

gccctgggta gggggtgggc tgggccctgg ggctcacaga cttgggcggt gtccctcctt  23340 

ggcatggggc ccgtgcctgc ctgtgggttc tcatctgtgt gcctgcatct gaccctcctg  23400 

tgcgcctgcg cctgaccctc ctgtgcgtgc ctgcccaggt gtggtgcgga tggaagcctt  23460 

gtctgctggc cggccagccc gaggccctga gcgcccagtg cccactgggg caaaggtgcc  23520 

tggagaaggc cccaggccag tgtctgcgac caccctgtga ggcctggggg gagtgcggcg  23580 

cagaagagcc accgagcacc ccctgcctgc cacgctccgg ccacctggac aataactgtg  23640 

cccgcctcac cttgcatttc aaccgtgacc acgtgcccca ggtgaggggc ctggtggcat  23700 

ctgagcttgc agaggccaca cgccggcatc tgctcgtggc atggcgaaag cctagccccg  23760 

cagggcaggg aggccctggt tggctgagca gagtcactct tggtcacaga gagtggccct  23820 

gtggggtcag atgagagggg cattgggcct ggtgctgggt ggaggtggca gaggaggctg  23880 

ggagagcagc cagctggggg tgcctgtttg tccagctgcc ctgagggcct ggactgacgg  23940 

cgccatggct gcctggcccc agctcttggg ctgcagctcc gtgggcagtt ttgccctggc  24000 

ctaggaccca cctttgcctg ctgtgtgctt ggagctgggc ccctgtctcc caggaggggc  24060 

tcagaactgg aggagaccca ctgtaccccg ccctgcctct ccttccccca ctggcctgca  24120 

ggtggagctg ggtccgccct gaggatgggc gggtgggcac cgtcactcct gcctcctggt  24180 

atagggcaca gccgggtggg aagctgcccc cccaggccct tggcatcctt gctgtgctct  24240 

cctgggcggg ctgtagggtg tgtcccacgt gtacccacag cgccagtcca gggatgtagg  24300 

tgtcaggttc acggccctgc cctgcccacg cactgcctgt ctctgcccag ggcaccacgg  24360 

tgggcgccat ttgctccggg atccgctccc tgccagccac aagggctgtg gcacgggacc  24420 

gcctgctggt gttgctttgc gaccgggcgt cctcgggggc cagtgctgtg gaggtggccg  24480 

tggtgagtgc ccagtgggga gcagcacctg ggtgggccct gggtcccgta ctatgcaggt  24540 

cctggctatg ctggacagag gctctggcga ggctagtcct ggtgcggaag gactgcgggc  24600 

aggcctgtct ccctgcggcc cctcgctgtc catgccgcag acccgtggaa ctgctccctg  24660 

ggcctggcca gcatgaggga gatgcagggc tgtggtgtgg agcccgcttc ccctgcagct  24720 

gcatcctcgc ccggtcccct gctctgtttt tgtctctgtg tccctacgtc acaggcagca  24780 

ggagagtccg tgggcttagt ctgccctggg aggcctgctt tgggactggc acctgccctg  24840 

gacctggggg gtgtcagatg tgaatggata ccaagggggt cgggtgagac tggggtggag  24900 

acatgcccgg agaggggagg gaatgttctg gaacatggtg ggtgggtgtg cagagcagtg  24960 

ggtgtggcca tggcacagtg tggctggtgg aggccatggc caggcacagg aaggacgtgc  25020 

agtgttttgg tgccctgagg ccgcagaggg ggtgggggac atggatgggt gctgctgggt  25080 

gatggaaggg cagtaggggc aggggaagat gtaagaagtg tgccagcaca ggtcagggcg  25140 

ccatcaggga tgtggtggag gcaggggcac agccccgggt tgctgtggcc tcgtgaaggc  25200 

actaggtttg tggtgcccct ggggtgtggc ccataggtgg gggtgggggc tgggaactga  25260 

caagaaggga tggccatcac ggagcaggtg tcagcgaatg gggccacaca cctccccaac  25320 

tcactgcctg gtggcgaggt ccccaccgca ggaccccggg ctctcctgtg tgcccggacg  25380 

gggacaccct ccacccctcc acttcccccc acccctcact gcctgctggt gaggtcccca  25440 

ccgcaggacc ctgggctgtc ccgtgcgccc ggatggggac atcctccacc cctccccttc  25500 

cccccactgc tcgctgcctg gtggtgaggt ccccacacct caggaccctg ggctctcctg  25560 

tgtgcccgga tggggacagc ctccacccct ccactcctcc ccccgctact ccccactcac  25620 

tgcctggtgg tgaagtcgcc actgcaggac cccgggctct cgtctcccgt gcgcccacct  25680 

tgctccagtg tggccagggc ctcagtgttg ggggcaggct gctgggagcc tggagccctc  25740 

gagccatccc cacaatgccg ttctttgccg cagtccttca gccctgccag ggacctgcct  25800 

gacagcagcc tgatccaggg cgcggcccac gccatcgtgg ccgccatcac ccagcggggg  25860 

aacagctcac tgctcctggc tgtcaccgag gtcaaggtgg agacggttgt tacgggcggc  25920 

tcttccacag gtaagcgcgg gaggtgggcc cctgggaagg caccaggcag gcaactcagg  25980 

cattgggcac agagccggcc gatcctgccg atcctgccag ccaccaggaa cacagaagtc  26040 

cctggcacct gctgccccag ccgcccagcc ccacaacctg accttcccag cccccgtcct  26100 

gggaccctcc ccacgagcca gcaaccggag ggtggggccc ggccgcctgg cccgcagggc  26160 

cctcccaggc ctgggtgtgt ggctagtgcc ccgcaggtgc ccaggcctca ttgcccaccg  26220 

gctcttctcc ccggtcccca ggtctgctgg tgcctgtgct gtgtggtgcc ttcagcgtgc  26280 

tgtggctggc gtgcgtggtc ctgtgcgtgt ggtggacacg caagcgcagg aaagagcggg  26340 

agaggagccg gctgccgcgg gaggagagcg ccaacaacca gtgggccccg ctcaacccca  26400 

tccgcaaccc cattgagcgg ccggggggcc acaaggacgt gctctaccag tgcaagaact  26460 

tcacgccgcc gccgcgcagg gcggacgagg cgctgcccgg gccggccggc cacgcggccg  26520 

tcagggagga tgaggaggac gaggatctgg gccgcggtga ggaggactcc ctggaggcgg  26580 

agaagttcct ctcacacaaa ttcaccaaag atcctggccg ctcgccgggg aggccggccc  26640 

actgggcctc aggccccaaa gtggacaacc gcgcggtcag gagcatcaat gaggcccgct  26700 

acgccggcaa ggagtagggg cggctgccag ctgggccggg acccagggcc ctcggtggga  26760 

gccatgccgt ctgccggacc cggaggccga ggccatgtgc atagtttctt tattttgtgt  26820 

aaaaaaacca ccaaaaacaa aaaccaaatg tttattttct acgtttcttt aaccttgtat  26880 

aaattattca gtaactgtca ggctgaaaac aatggagtat tctcggatag ttgctatttt  26940 

tgtaaagttt ccgtgcgtgg cactcgctgt atgaaaggag agagcaaagg gtgtctgcgt  27000 

cgtcaccaaa tcgtagcgtt tgttaccaga ggttgtgcac tgtttacaga atcttccttt  27060 

tattcctcac tcgggtttct ctgtggctcc aggccaaagt gccggtgaga cccatggctg  27120 

tgttggtgtg gcccatggct gttggtggga cccgtggctg atggtgtggc ctgtggctgt  27180 

cggtgggact cgtggctgtc aatgggacct gtggctgtcg gtgggaccta cggtggtcgg  27240 

tgggaccctg gttattgatg tggccctggc tgccggcacg gcccgtggct gttgacgcac  27300 

ctgtggttgt tagtggggcc tgaggtcatc ggcgtggccc aaggccggca ggtcaacctc  27360 

gcgcttgctg gccagtccac cctgcctgcc gtctgtgctt cctcctgccc agaacgcccg  27420 

ctccagcgat ctctccactg tgctttcaga agtgcccttc ctgctgcgca gttctcccat  27480 

cctgggacgg cggcagtatt gaagctcgtg acaagtgcct tcacacagac ccctcgcaac  27540 

tgtccacgcg tgccgtggca ccaggcgctg cccacctgcc ggccccggcc gcccctcctc  27600 

gtgaaagtgc atttttgtaa atgtgtacat attaaaggaa gcactctgta tatttgattg  27660 

aataatgcca ccattccggc ctcccttgtt ctttcggtgc tgtccctttt gtattgagag  27720 

tgaggttggg ggagagccac gccggcagag aggcttgggg cagtggggca cgtgctgggt  27780 

attggcccac gtggctgtgg tggctgtaga gggcgagacg gttctgttga gtcggggcct  27840 

gccagggcct cgaatgcgtt ggcatgccaa ggtggtggat gcaggtttgg ccaaaacctt  27900 

cctgggaatg gggagggggg tgtctaggtg cctggcaccc gaccctgact aaaacagctg  27960 

aaaacagttt tataaaatag tataaaattg cttacccacg                        28000  
           
             382  
             419  
             DNA  
             Homo sapiens  
             
               3′UTR  
               (1)...(419)  
             
           
            382 

tgcggccgcc ccttctcgtg aaagtgcatt tttgtaaatg tgtacatatt aaaggaagca     60 

ctctgtatat ttgattgaat aatgccacca ttccggcctc ccttgttctt tcggtgctgt    120 

cccttttgta ttgagagtga ggttggggga gagccacgcc ggcacatagg cttggggcag    180 

tggggcacgt gctgggtatt ggcccacgtg gctgtggtgg ctgtataggg cgagaccgat    240 

ctgttgagtc ggggcctgcc acggcctcga atgcgttggc atgccaaggt ggtggatgca    300 

ggtttggcct aaaccttcct gagaatgggg acgggggtgg atctggaatt ggcatgatta    360 

caaactactc tgcaattctt cctctcccca attaaggtgt ctctcttgaa ctgattgaa     419 

 
           
             383  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            383 

tacaaaaatg cactttcacg                                                 20 

 
           
             384  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            384 

tggcattatt caatcaaata                                                 20 

 
           
             385  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            385 

gcgcacctgc atatgcatga                                                 20 

 
           
             386  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            386 

gaaatagccc atgggccgcg                                                 20 

 
           
             387  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            387 

cagctgcagc tcgaaatagc                                                 20 

 
           
             388  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            388 

gcagcgcgct cagctgcagc                                                 20 

 
           
             389  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            389 

gcagctcccc gttcacgttc                                                 20 

 
           
             390  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            390 

gctcagcagc tccccgttca                                                 20 

 
           
             391  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            391 

tggtactcct taaggcacac                                                 20 

 
           
             392  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            392 

caccttggcc tggtactcct                                                 20 

 
           
             393  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            393 

gccgtagctg cagggccccg                                                 20 

 
           
             394  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            394 

ggcaggtaga aggagttgcc                                                 20 

 
           
             395  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            395 

gacgaggccc gggtcctggt                                                 20 

 
           
             396  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            396 

ttgtcccagt cccaggcctc                                                 20 

 
           
             397  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            397 

aggctcttcc agcggtcctc                                                 20 

 
           
             398  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            398 

gctgaagtgc aggctcttcc                                                 20 

 
           
             399  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            399 

ccacgtggcc gctgaagtgc                                                 20 

 
           
             400  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            400 

ggccggcaga acttgttgca                                                 20 

 
           
             401  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            401 

ttgccgtact ggtcgcaggt                                                 20 

 
           
             402  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            402 

gcaggccttg ttgccgtact                                                 20 

 
           
             403  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            403 

catccagccg tccatgcagg                                                 20 

 
           
             404  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            404 

cccccgtgga gcaaattaca                                                 20 

 
           
             405  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            405 

gtagctgcac ctgcactccc                                                 20 

 
           
             406  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            406 

cagttgcact gccagggctc                                                 20 

 
           
             407  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            407 

gttggtctca cagttgcact                                                 20 

 
           
             408  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            408 

ccgccccagt tggtctcaca                                                 20 

 
           
             409  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            409 

gcaggccgcc ccagttggtc                                                 20 

 
           
             410  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            410 

acagagcagg ccgccccagt                                                 20 

 
           
             411  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            411 

ttgtcacaga gcaggccgcc                                                 20 

 
           
             412  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            412 

ttcaggtctt tgtcacagag                                                 20 

 
           
             413  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            413 

gccacagtag ttcaggtctt                                                 20 

 
           
             414  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            414 

ggtggtggct gccacagtag                                                 20 

 
           
             415  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            415 

gaggtgcagg cgtgctcagc                                                 20 

 
           
             416  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            416 

cccttcacac tcattggcgt                                                 20 

 
           
             417  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            417 

aggtttttgc aagaaaaagc                                                 20 

 
           
             418  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            418 

cacagtaata gccgccaatc                                                 20 

 
           
             419  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            419 

gatgcccttc cagcccggga                                                 20 

 
           
             420  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            420 

gcaggtgccc ccatgctgac                                                 20 

 
           
             421  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            421 

ccaggtcctt gcaggtgccc                                                 20 

 
           
             422  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            422 

gggcacacac actggtaccc                                                 20 

 
           
             423  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            423 

gggctgctgg cacacttgtc                                                 20 

 
           
             424  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            424 

gagcagttct tgccaccaaa                                                 20 

 
           
             425  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            425 

ccgcagccat cgatcactct                                                 20 

 
           
             426  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            426 

gtgcccccat tgcggcaggg                                                 20 

 
           
             427  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            427 

agaagtcatt gaccaggtcg                                                 20 

 
           
             428  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            428 

cacagtagaa gtcattgacc                                                 20 

 
           
             429  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            429 

cgtgagtggc aggtcttgcc                                                 20 

 
           
             430  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            430 

ctggaactcg cgtgagtggc                                                 20 

 
           
             431  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            431 

ccgttgctgc aggtgtaggc                                                 20 

 
           
             432  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            432 

caggtgccac cgttgctgca                                                 20 

 
           
             433  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            433 

cgtagcaggt gccaccgttg                                                 20 

 
           
             434  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            434 

ttgggcaggc agctgctgtt                                                 20 

 
           
             435  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            435 

agggttgcag tcgttggtat                                                 20 

 
           
             436  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            436 

gcagcggaac cagttgacgc                                                 20 

 
           
             437  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            437 

ccgtaggcac agggcgagga                                                 20 

 
           
             438  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            438 

ttgatctcat ccacacacgt                                                 20 

 
           
             439  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            439 

ggtgggcagc tacagcgata                                                 20 

 
           
             440  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            440 

gcagctgttg cagtcttcca                                                 20 

 
           
             441  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            441 

ccaggcagcg gcagctgttg                                                 20 

 
           
             442  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            442 

ctgctgtcag gcaggtccct                                                 20 

 
           
             443  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            443 

ctggatcagg ctgctgtcag                                                 20 

 
           
             444  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            444 

tccaccttga cctcggtgac                                                 20 

 
           
             445  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            445 

gcgcggttgt ccactttggg                                                 20 

 
           
             446  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            446 

ccctactcct tgccggcgta                                                 20 

 
           
             447  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            447 

gacggcatgg ctcccaccga                                                 20 

 
           
             448  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            448 

gaataattta tacaaggtta                                                 20 

 
           
             449  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            449 

aatactccat tgttttcagc                                                 20 

 
           
             450  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            450 

tcatacagcg agtgccacgc                                                 20 

 
           
             451  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            451 

caccctttgc tctctccttt                                                 20 

 
           
             452  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            452 

caccggcact ttggcctgga                                                 20 

 
           
             453  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            453 

gggtcccacc aacagccatg                                                 20 

 
           
             454  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            454 

gaagggcact tctgaaagca                                                 20 

 
           
             455  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            455 

acagttccga gggttctgtg                                                 20 

 
           
             456  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            456 

ctggctggat cccccacact                                                 20 

 
           
             457  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            457 

gggagcactc ctggctctgc                                                 20 

 
           
             458  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            458 

ccatactgac tgatatggca                                                 20 

 
           
             459  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            459 

cgacatccac ctgcagggtg                                                 20 

 
           
             460  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            460 

tggcaggccc cgactcaaca                                                 20 

 
           
             461  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            461 

nnnnnnnnnn nnnnnnnnnn                                                 20 

 
           
             462  
             2574  
             DNA  
             Homo sapiens  
             
               CDS  
               (254)...(1492)  
             
           
            462 

cctgtttaga cacatggaca acaatcccag cgctacaagg cacacagtcc gcttcttcgt     60 

cctcagggtt gccagcgctt cctggaagtc ctgaagctct cgcagtgcag tgagttcatg    120 

caccttcttg ccaagcctca gtctttggga tctggggagg ccgcctggtt ttcctccctc    180 

cttctgcacg tctgctgggg tctcttcctc tccaggcctt gccgtccccc tggcctctct    240 

tcccagctca cac atg aag atg cac ttg caa agg gct ctg gtg gtc ctg       289 
               Met Lys Met His Leu Gln Arg Ala Leu Val Val Leu 
                1               5                   10 

gcc ctg ctg aac ttt gcc acg gtc agc ctc tct ctg tcc act tgc acc      337 
Ala Leu Leu Asn Phe Ala Thr Val Ser Leu Ser Leu Ser Thr Cys Thr 
         15                  20                  25 

acc ttg gac ttc ggc cac atc aag aag aag agg gtg gaa gcc att agg      385 
Thr Leu Asp Phe Gly His Ile Lys Lys Lys Arg Val Glu Ala Ile Arg 
     30                  35                  40 

gga cag atc ttg agc aag ctc agg ctc acc agc ccc cct gag cca acg      433 
Gly Gln Ile Leu Ser Lys Leu Arg Leu Thr Ser Pro Pro Glu Pro Thr 
 45                  50                  55                  60 

gtg atg acc cac gtc ccc tat cag gtc ctg gcc ctt tac aac agc acc      481 
Val Met Thr His Val Pro Tyr Gln Val Leu Ala Leu Tyr Asn Ser Thr 
                 65                  70                  75 

cgg gag ctg ctg gag gag atg cat ggg gag agg gag gaa ggc tgc acc      529 
Arg Glu Leu Leu Glu Glu Met His Gly Glu Arg Glu Glu Gly Cys Thr 
             80                  85                  90 

cag gaa aac acc gag tcg gaa tac tat gcc aaa gaa atc cat aaa ttc      577 
Gln Glu Asn Thr Glu Ser Glu Tyr Tyr Ala Lys Glu Ile His Lys Phe 
         95                 100                 105 

gac atg atc cag ggg ctg gcg gag cac aac gaa ctg gct gtc tgc cct      625 
Asp Met Ile Gln Gly Leu Ala Glu His Asn Glu Leu Ala Val Cys Pro 
    110                 115                 120 

aaa gga att acc tcc aag gtt ttc cgc ttc aat gtg tcc tca gtg gag      673 
Lys Gly Ile Thr Ser Lys Val Phe Arg Phe Asn Val Ser Ser Val Glu 
125                 130                 135                 140 

aaa aat aga acc aac cta ttc cga gca gaa ttc cgg gtc ttg cgg gtg      721 
Lys Asn Arg Thr Asn Leu Phe Arg Ala Glu Phe Arg Val Leu Arg Val 
                145                 150                 155 

ccc aac ccc agc tct aag cgg aat gag cag agg atc gag ctc ttc cag      769 
Pro Asn Pro Ser Ser Lys Arg Asn Glu Gln Arg Ile Glu Leu Phe Gln 
            160                 165                 170 

atc ctt cgg cca gat gag cac att gcc aaa cag cgc tat atc ggt ggc      817 
Ile Leu Arg Pro Asp Glu His Ile Ala Lys Gln Arg Tyr Ile Gly Gly 
        175                 180                 185 

aag aat ctg ccc aca cgg ggc act gcc gag tgg ctg tcc ttt gat gtc      865 
Lys Asn Leu Pro Thr Arg Gly Thr Ala Glu Trp Leu Ser Phe Asp Val 
    190                 195                 200 

act gac act gtg cgt gag tgg ctg ttg aga aga gag tcc aac tta ggt      913 
Thr Asp Thr Val Arg Glu Trp Leu Leu Arg Arg Glu Ser Asn Leu Gly 
205                 210                 215                 220 

cta gaa atc agc att cac tgt cca tgt cac acc ttt cag ccc aat gga      961 
Leu Glu Ile Ser Ile His Cys Pro Cys His Thr Phe Gln Pro Asn Gly 
                225                 230                 235 

gat atc ctg gaa aac att cac gag gtg atg gaa atc aaa ttc aaa ggc     1009 
Asp Ile Leu Glu Asn Ile His Glu Val Met Glu Ile Lys Phe Lys Gly 
            240                 245                 250 

gtg gac aat gag gat gac cat ggc cgt gga gat ctg ggg cgc ctc aag     1057 
Val Asp Asn Glu Asp Asp His Gly Arg Gly Asp Leu Gly Arg Leu Lys 
        255                 260                 265 

aag cag aag gat cac cac aac cct cat cta atc ctc atg atg att ccc     1105 
Lys Gln Lys Asp His His Asn Pro His Leu Ile Leu Met Met Ile Pro 
    270                 275                 280 

cca cac cgg ctc gac aac ccg ggc cag ggg ggt cag agg aag aag cgg     1153 
Pro His Arg Leu Asp Asn Pro Gly Gln Gly Gly Gln Arg Lys Lys Arg 
285                 290                 295                 300 

gct ttg gac acc aat tac tgc ttc cgc aac ttg gag gag aac tgc tgt     1201 
Ala Leu Asp Thr Asn Tyr Cys Phe Arg Asn Leu Glu Glu Asn Cys Cys 
                305                 310                 315 

gtg cgc ccc ctc tac att gac ttc cga cag gat ctg ggc tgg aag tgg     1249 
Val Arg Pro Leu Tyr Ile Asp Phe Arg Gln Asp Leu Gly Trp Lys Trp 
            320                 325                 330 

gtc cat gaa cct aag ggc tac tat gcc aac ttc tgc tca ggc cct tgc     1297 
Val His Glu Pro Lys Gly Tyr Tyr Ala Asn Phe Cys Ser Gly Pro Cys 
        335                 340                 345 

cca tac ctc cgc agt gca gac aca acc cac agc acg gtg ctg gga ctg     1345 
Pro Tyr Leu Arg Ser Ala Asp Thr Thr His Ser Thr Val Leu Gly Leu 
    350                 355                 360 

tac aac act ctg aac cct gaa gca tct gcc tcg cct tgc tgc gtg ccc     1393 
Tyr Asn Thr Leu Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val Pro 
365                 370                 375                 380 

cag gac ctg gag ccc ctg acc atc ctg tac tat gtt ggg agg acc ccc     1441 
Gln Asp Leu Glu Pro Leu Thr Ile Leu Tyr Tyr Val Gly Arg Thr Pro 
                385                 390                 395 

aaa gtg gag cag ctc tcc aac atg gtg gtg aag tct tgt aaa tgt agc     1489 
Lys Val Glu Gln Leu Ser Asn Met Val Val Lys Ser Cys Lys Cys Ser 
            400                 405                 410 

tga gaccccacgt gcgacagaga gaggggagag agaaccacca ctgcctgact          1542 

gcccgctcct cgggaaacac acaagcaaca aacctcactg agaggcctgg agcccacaac   1602 

cttcggctcc gggcaaatgg ctgagatgga ggtttccttt tggaacattt ctttcttgct   1662 

ggctctgaga atcacggtgg taaagaaagt gtgggtttgg ttagaggaag gctgaactct   1722 

tcagaacaca cagactttct gtgacgcaga cagaggggat ggggatagag gaaagggatg   1782 

gtaagttgag atgttgtgtg gcaatgggat ttgggctacc ctaaagggag aaggaagggc   1842 

agagaatggc tgggtcaggg ccagactgga agacacttca gatctgaggt tggatttgct   1902 

cattgctgta ccacatctgc tctagggaat ctggattatg ttatacaagg caagcatttt   1962 

tttttttaaa gacaggttac gaagacaaag tcccagaatt gtatctcata ctgtctggga   2022 

ttaagggcaa atctattact tttgcaaact gtcctctaca tcaattaaca tcgtgggtca   2082 

ctacagggag aaaatccagg tcatgcagtt cctggcccat caactgtatt gggccttttg   2142 

gatatgctga acgcagaaga aagggtggaa atcaaccctc tcctgtctgc cctctgggtc   2202 

cctcctctca cctctccctc gatcatattt ccccttggac acttggttag acgccttcca   2262 

ggtcaggatg cacatttctg gattgtggtt ccatgcagcc ttggggcatt atgggtcttc   2322 

ccccacttcc cctccaagac cctgtgttca tttggtgttc ctggaagcag gtgctacaac   2382 

atgtgaggca ttcggggaag ctgcacatgt gccacacagt gacttggccc cagacgcata   2442 

gactgaggta taaagacaag tatgaatatt actctcaaaa tctttgtata aataaatatt   2502 

tttggggcat cctggatgat ttcatcttct ggaatattgt ttctagaaca gtaaaagcct   2562 

tattctaagg tg                                                       2574 

 
           
             463  
             22  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            463 

accaattact gcttccgcaa ct                                              22 

 
           
             464  
             24  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            464 

gatcctgtcg gaagtcaatg taga                                            24 

 
           
             465  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            465 

aggagaactg ctgtgtgcgc ccc                                             23 

 
           
             466  
             29000  
             DNA  
             Homo sapiens  
           
            466 

ttatctctaa gatcctttgt taacttgctt ctcagtgaga atactttgtt agattttaat     60 

ctgaagaaac actcattgcc aaaaatccaa gacagatgaa atataggctc caatgaatcc    120 

cttgttccct tcttcttcgg caccagatga ggaagaaggg attaatcttt ggatttttcc    180 

atttcttgta agctcacagc atacgaccat caccttatag tcagtttcca gaccttcacc    240 

cccacttctt ccccaacttg ctgaaaacag aaggcaaatg gtcctcactc tgggcagaga    300 

ggtaccctgc agtagtagct tccagaactt gcttagcacc tgaatcacgt gtgaggtttg    360 

taaagaaaca gagatgccag ggcctcagct ctggagactg atttggtaga ggtggagtcc    420 

aaaaaagtat aactttaata attttccttc ctatcttgca actgtctgct caaaggcctt    480 

cccttatcac cctatttgaa actgcaacat cccccaacct aggcacaccc catcctcctt    540 

ccctgcttga ttttctgcca caccacattt gtttgtttgc ttgtctgttt gagacacggt    600 

cttgctctgt cgtccaggct ggagtgcagt ggtgcaatct tggccccctg taaactctgc    660 

ctccctggct caagtgatta tcctgcctca gcctcccaag tagatgcctg cgccaacatg    720 

ccgggctaat ttttccattt ttttgtagag actgggtttc gccgtgttgc tggggctggt    780 

ctcgaattcc tgagctcaag taatcctcct gcatgggcct ccccaaatgc tgggattaca    840 

ggcgtgagcc actgcacctg gctcagcact ttttaccgta ctacatcatt tacatattta    900 

tttagtttat cgcctcctcc actgccccac ccctgcctct aaaataaaat ttccctgagg    960 

gcaggagttt tgtttcgttc actgatattc ttcacagagc ctagaatagt gcctggtata   1020 

tagtaaacat taaacttttt ctgaaatttc agaggcagta tagcatagta attaagtcca   1080 

gaatctggca acgtcctggg tccaaatccc aacagctgac acctaataac tatgtgacct   1140 

tgggcaagtt acttttaaag tttctacccc taggtttccc attggttttg caaatgaaag   1200 

taatgcctac ccaagctaga tagcctgtgt aaaatatcgc ctccatcact cacaagcagt   1260 

gtggtctgta aaaaaaaaaa caaaaaactc tatgcctcag tttcctcatc cgtaaagtga   1320 

cccaccgctg tgctgggata cagagaacag ccccttcagt tagtggcctg gaagccagac   1380 

ctctcagaaa gggtccagga aggctggagt gagatggggt gggagcggca ctcactctca   1440 

ggaaagttca gttcagaggc aagccctgtg ttgcggggtg cggggagcca cgtgccctac   1500 

cctcccttgg ctgctcgtgg gaaaaggcct agaggttcgg gccgagaaga ggagcgaaag   1560 

cacagagccg acttcccctc acccatctgg gaaatgggct cgggccaact gctgacttcg   1620 

cgctcgctgg cgcagctccc tgcggagacc tcggcgggga gggaggctga acatctggat   1680 

gacatttctg cgagagcggc tccggagcgg cggtcgggga gggagaggtg cgctcgtgcg   1740 

cacgtcgggc cgggagggag gcgattcctc ggggcctggg tcttgttttt ctcgctctct   1800 

accgcagccc cttctcccgc ccctcagccc ccaccccgca gcccccagcc cccgagcctc   1860 

cccggctccc gaccagccga gctccttcac tggcggcctc gcctcgccag agggcaccct   1920 

cgatcttccg gaaaacgcca ccatttttca ctgcccctgg agcgtctcca ggcttctgcc   1980 

cgcctcccga ctccgatctt gtcaatgaag aatcgggcca ggatcgccgc ggagcggacg   2040 

ccgaccctcc gacccggctc gcaggctggg agtcccctct gcgaggctgg catggccgcc   2100 

cctaccgggt cccgcgccct ctgcggaccc tgccccgggt tgggcctggc ccgcgggcgg   2160 

ccccgggacc gggggaccag gagggagagt agacgcgggc cgcggacggc gcggactgac   2220 

agctggcgag agggcgccgg ggctggggga aagggaggga gggggctcat cggagtaact   2280 

ttccagaaaa acagccaacg tgtggcagga gtgattccaa gaggggaaaa aaagttcagc   2340 

taccacgtcg aacgagagga ctcgcaaagt atttttcaaa agggctcggc ttttcctgtg   2400 

cctgtttaaa acattaacat cgtgcagcaa aagaggctgc gtgcgctggt ccctccctcc   2460 

cccaccccag gccagagacg tcatgggagg gaggtataaa atttcagcag agagaaatag   2520 

agaaagcagt gtgtgtgcat gtgtgtgtgt gtgagagaga gagggagagg agcgagaggg   2580 

agagggagag ggagagagag aaagggaggg aagcagagag tcaagtccaa gggaatgagc   2640 

gagagaggca gagacagggg aagaggcgtg cgagagaagg aataacagct ttccggagca   2700 

ggcgtgccgt gaactggctt ctattttatt ttattttttt ctccttttta ttttttaaag   2760 

agaagcaggg gacagaagca atggccgagg cagaagacaa gccgaggtgc tggtgaccct   2820 

gggcgtctga gtggatgatt ggggctgctg cgctcagagg cctgcctccc tgccttccaa   2880 

tgcatataac cccacacccc agccaatgaa gacgagaggc agcgtgaaca aagtcattta   2940 

gaaagccccc gaggaagtgt aaacaaaaga gaaagcatga atggagtgcc tgagagacaa   3000 

gtgtgtcctg tactgccccc acctttagct gggccagcaa ctgcccggcc ctgcttctcc   3060 

ccacctactc actggtgatc tttttttttt tacttttttt tcccttttct tttccattct   3120 

cttttcttat tttctttcaa ggcaaggcaa ggattttgat tttgggaccc agccatggtc   3180 

cttctgcttc ttctttaaaa tacccacttt ctccccatcg ccaagcggcg tttggcaata   3240 

tcagatatcc actctattta tttttaccta aggaaaaact ccagctccct tcccactccc   3300 

agctgccttg ccacccctcc cagccctctg cttgccctcc acctggcctg ctgggagtca   3360 

gagcccagca aaacctgttt agacacatgg acaagaatcc cagcgctaca aggcacacag   3420 

tccgcttctt cgtcctcagg gttgccagcg cttcctggaa gtcctgaagc tctcgcagtg   3480 

cagtgagttc atgcaccttc ttgccaagcc tcagtctttg ggatctgggg aggccgcctg   3540 

gttttcctcc ctccttctgc acgtctgctg gggtctcttc ctctccaggc cttgccgtcc   3600 

ccctggcctc tcttcccagc tcacacatga agatgcactt gcaaagggct ctggtggtcc   3660 

tggccctgct gaactttgcc acggtcagcc tctctctgtc cacttgcacc accttggact   3720 

tcggccacat caagaagaag agggtggaag ccattagggg acagatcttg agcaagctca   3780 

ggctcaccag cccccctgag ccaacggtga tgacccacgt cccctatcag gtcctggccc   3840 

tttacaacag cacccgggag ctgctggagg agatgcatgg ggagagggag gaaggctgca   3900 

cccaggaaaa caccgagtcg gaatactatg ccaaagaaat ccataaattc gacatgatcc   3960 

aggggctggc ggagcacagt aagtccaaat tctcgctggg gtgtctgctc tggagggtct   4020 

gaactggagc tgggagctct gcagaggggg gcctagtgct ggccacacag cagggtgccc   4080 

caggattcac cagcaccaag gctcaggatg tgcgatgctc ctccgttggg gctggggagg   4140 

tgggtgggga aggagataga gccattctgt taagagccgg cgcttctggg aggccaggag   4200 

ccctggagct gagtggcttg ctgaattcac atcacatcct tgactgattt taatttggaa   4260 

ttacattgtg ctgtccaggg aaacatatgt attcttgcac atgcgatcgt atcagtaact   4320 

gtaagcatct gggtgccata aaggggaagg ccggctctgt caggagccct tacggttctc   4380 

agtgtggaga cctcatcttc tccctgcttt tcacaactca ttgtgacacg tctccgtttc   4440 

agtttttcca gttcttggga agaagaatac ctgccccaaa ttaatgtctg tcaagctttt   4500 

tgaagcccag gcaggagaca gcttcttgct gcctgggccc tttggtctac cccacccacg   4560 

tgacccacga gacccacgtg agctgtgtgt gtggaaggaa gagggtatgc acgaatgttc   4620 

ccagggccgt gtactttagg gtgacatgca gtcttgtgca gtagacagat tcatgtgctc   4680 

aaaatgggcg ccctccaggc cggtgggcac ggggagagcg ggttttggct gtggatgcgt   4740 

agaggaggct ggcgcccttt gtgtctgcgt gtcacgggag agcgggtgga ggggtggcag   4800 

tgggtgcatg gtgggggggg gggatatgtc tgggagcctg ccgtcccagg aggctctgtc   4860 

tgcatggagg agccgggcgg cttctgggcg agatgtctgt gtgtgttggt acacgtgtgg   4920 

aagtcatatg tgtttactga aggggatttt aaaaacctca atacaagaga gagaaatttg   4980 

gcagatgttg agaaactgac agcccaggaa agaggaatgt gagccactcg tgggccgtag   5040 

actccgggag cagctctgtt tgcttttcct accagcaggt gtcctcgccg ccctgactac   5100 

ctcagcccag gcccacctgg gaggtgggca gctcctggag tggggtggag ggcatgggat   5160 

ggagctggca ggcaggggag ggtggtcagc agagcacaca gcaaggggtg aaaggaacct   5220 

ggctggagag aaggaacagg agtgggtacc gatgggtgga ccagctctgg ctggaggtgc   5280 

aaaggccccg ttcacggctc cacgccaggc agaggagcct gtggttactg gcgaggggtt   5340 

cccgctccag cttcctgtgg ctgcctggag cgcctttctt caggatgtgg ctgccatgtg   5400 

gggcggaggc tggaggccga tgcagagcta ctactccctg cccagggtct ctgggtgggg   5460 

ctggctcaga gacccacagt tcccagaggc acctagcagc tcgatggcca aggctccaac   5520 

tccctgggaa cccaccaacg cgggagatag tgaccacaag catcagagga aggtcgaaat   5580 

ctgaggccgg caggagaggt gtgaggagag tccagggcaa gagggcagga ctcagacctt   5640 

catggtctgg gtcagcagga ggagtccaag ggaggaagca ttctgagtca ccaggacccc   5700 

cccatcccgg aatcctgagc tgagaatgaa tgagccacgt ggaggcaagg ccatgcaggt   5760 

gcaagtggac actgattttg tgcagactca aagcacaaat agcagatgtc cttgggaaaa   5820 

gcccgggcag ggccccatag atgctgggca gcttccaggc tgcagtacca agaccttaca   5880 

actgcaacag atgggtggat gtggggttat ggagcaatgg tctggcctgg ggcaacccag   5940 

cacagtgagc aggatgctgt tcaggatgct ggggaggagc caacgtgcga tgctatgagg   6000 

ctcacaggta caaaccggaa gcaggcagac tctgcagctg ttggaggtga cttggaggct   6060 

gagcagacgg acctgggccc gccctgcagc tggtcgggtg ctgagcccac cccagagagg   6120 

cagacacaca aggcacacta actataaaga aggcagtggg caggtgctga gcaggagcag   6180 

agagccatca tcaggggctt gcaaggcggc ggggcggggt gggggaagga agcctgtctt   6240 

taactcatga gggcagacag gggtgacacc aggtctgtgg tggggcacag cagggtctca   6300 

atgccagagc ctctgctggg aggtcatgag atcacgttct gttccatatt tcctcacttc   6360 

tggccacttc cctgacccag tgaacatgca ttcaaaggaa agtgacagta ggagccaggg   6420 

caaggagata gaggtccctg gagaggaaaa tgaaagagga aatacttttt agtagtgcag   6480 

gagaaagggc accaaggtga gagcagagag gaaggccttt tcctaaataa ccttttctcc   6540 

ctgttttaca gataaggaaa ctgagacctg gattgcttaa gtaatttgtc caaaaagagc   6600 

agggacccta acctagacat tctgtgtgca ggacgcatgt agttaagcac tcatttatat   6660 

gtaaaataca cgttgtaagt gttaccttta acctccttta acctttaggg ttcagtagaa   6720 

ctttgattta taacataaat gaatcatgtg ttggaccaag caggagaggt cagagttatt   6780 

atcttagtaa cccaggtggc agattgcaca atgataactg gatttgttct tccttagctc   6840 

tgcatttttt tttttttttt tttgcccggg agattcatac tgccacaaat gttctcccta   6900 

atttaatgaa ggagttttct ttatttaatg aagagtctca agcaggttaa gcaaccgcag   6960 

ctacgtaaaa gtgacctctc tgagcctcag tttccccagc tgtaaagtta gagatgattt   7020 

ccaaactcct tttcagctga aagaatttta taattccatc tgggatgaat cagcagagcc   7080 

tctattgggg agtatgggca agactctgta atcctttttc taattctcca ggattttact   7140 

gtcgggaggg agtagagagt ttctctgacc ccatgtgatg ggaaaggaca cagctttttt   7200 

acttccgttg tcatccctct tacaaaggta tcaccaatgt aggtgtcatt ttatcttctg   7260 

gcttgtaatt atctgtctct gttcggagac ttgttgtttt cagccaaggg cagcgctaag   7320 

acaaccagca aacccagagt ttctcagcaa agagaaaact ctatatttta gtctttgttc   7380 

tctagctgct aagtgtagat tttgtttatt ctgagaatta ttctgaaaat catttgctcc   7440 

aagggccaat gccctctgca cagtagaggt cagcacttct ccaagtgtgg tccagaggaa   7500 

gctggaggta aatgtagatt ccctggtccc acccaccctt atagaatcag aatcttgagg   7560 

gggtggagtc ttggggaacc tgtattttcg acaagctcct tatggattct taagcacatt   7620 

gaagcttaag agtcagtgaa ctagggcgaa acttttctta gagggatggc aaacacaagt   7680 

gcctacagag acccggcagg aaatgcaaat gatctggaag aaaagccacg gcgtcatgat   7740 

aaactgcacc aggacacttg gtcttggggt caagaagaaa gtagggtgtg tgagacaggg   7800 

agagggaggg gacctggagc ccacgtgccc agccaaagca gcagccagcc tcagttcttg   7860 

ctgggttttg cattgaggac tgtgggtcca gcttgattag ttcttcccgt gtcccagaaa   7920 

agcagaaaat ctggatcttt ctgtgaagtg ttccaatttt taacatgggc ttaaaatgtt   7980 

tatgggcttc taactcaaaa tttttaaagg tgttccatca gcgaaacaac atgtctaatt   8040 

catttaacgg ttaatcaata gaaagctcac accattaaag cagtggtttc tcaaacttcc   8100 

agaacatcta gaagccatgg tgccctttgc aacacattat aatctgtggt tctcaaccct   8160 

ggctgcacgt tagaatcatc tggagatctt gaaaaaaata tgccgtggac cccactcact   8220 

ccagtgtagt cagaactgct ggggaatggg tccaggaatc atttgttttt aaagctttcc   8280 

aggtgattct aatgtgcagc cagggtagag aagtacagcc acactgataa atatagtccc   8340 

ttcactagaa ccagcagaaa tgatatatac aagcaaaggc acacctagcc acccaggtgt   8400 

ctgaacacat tttaaaaggc agttaactaa acatggtcag ctatttcctg gtttttccat   8460 

gcatactgta catgaatatt ctttgcttat gttttgcccc gttaaacaaa atagagaaaa   8520 

tggcattcac caatatatat tttttctgtg caatggaaaa agttgctcaa gatttaattt   8580 

gtaaaggtgg gaccccctag tccagctctc aataatacta gtgttctgtg aggcatggct   8640 

taagaaccac aaactcgttt gcagtgggtc attgtctgag gcataggttg acactctagg   8700 

cccatttagt ggaatcttgc catattttgt tgatgaaacc atcttcacca gatgatctcc   8760 

cagatccctc ccagcttgaa gagtctctgc ttcaataaat gaggtatgtt cagaagacct   8820 

gggttcaaac cccacctcca ccaccttcta gttatgtgac cttgggaaag acatttaact   8880 

ttttgaggct cagttttctc atttgtcaag tgataaattt tacatgtttt cattcttctc   8940 

aggggttgtt agaaggtcac atgaagtaat aaaaactcga caaaacaagg tggtgctatt   9000 

acattttgct tatttatgta tacgatgatt cattccacag attacttaaa acatcattat   9060 

tcagtgaatt tgattgtcaa gaagattgta tgtacatttt ctttgatctc ccaggcaatt   9120 

ctttttttta attaatttta attttaattt ttttgacaga gtctcactct gtcacccagc   9180 

ctggagtgca ctggtgcaat ctcggctcac tgtagcttct gccttctggg ttcaagtgat   9240 

tctcatgcct cagcctcccg agtagctggg gttacaggtg cccaccacca cacccagcta   9300 

atttttgtat ttggagtaga gatggggttt tgccatgttg gccaggctgg tctcgaactc   9360 

ctgacctcca gtgatccacc tgcctcggcc tcccaaagtg ctgggattac aggcatgagc   9420 

cactgttctc ggcctttaaa atttttaatt ttaaataata gggataggtc ctccctatgt   9480 

tgtccaggct gatcttgaac tcctgggctc aagcaatcct cccgcctcag cctcccgagt   9540 

agctgaaata acagacatgt gctaccatgc ccagctaatt ttcgtatttt ttatagagat   9600 

ggggtttcac catgttggcc aggctggtct caaacacctg agctcaagca atccacccac   9660 

ctcagcctcc caaggtgctg gctgggatta caggcgtgag ccaccatgcc tggctgccaa   9720 

ttcttcttct tcttcttctt tttttttttt tttttgagat ggagtctcac tctgttgccc   9780 

aggctggagt gcagtgacac aatctcagct cactgcaacc tcgacctccc aggttcaagt   9840 

gactctcctg cctcagcctc ctgaatagct aggattacaa gcatgcacca tcatgcctgg   9900 

ctaatttttg tatttttagt agagacgggg tttcaccttg ttgcccaggt gccaattctt   9960 

ttttaatcac tagcaattgt gtcctaagct ttgcttgcta gtgtcaagtt gcttgtgtca  10020 

gctaacttct gagtgactct ggccaagacc ctctagacag ccatttcttc ctctgaagag  10080 

ggttgcgcca catgactcct gatgtccctt ctaatcatgg gaaatctata tatcccagta  10140 

atagaaaaat gacctttccc acctctttct tgaaacctta aaattctccc caggatgtgt  10200 

tcatcctggg gagcagatta tgattgatag gctggaagaa accaaagagg acggccacta  10260 

gggtgtcctg agaactctct tagctcataa ctttccccat ctcctggctt cccactgcct  10320 

tgacccactc tgactgtctc accagcaagt gccattttcc atctcccttc ttttttttct  10380 

gagatggagt ctcactctgt tgcccaggtt ggagtgcaat ggcaccacct cagctcattg  10440 

caacctctgc ctcctgggtt caagcgattc tcctgcctca acctcctcag tagctgggat  10500 

tacaggcacg caccaccagg cctggctaat tttcatattt ttagtagaga cagggtttca  10560 

ccatgttggt caggctggtc tcaaactcct gatgtcgtga tctgcccacc tcggcttccc  10620 

gaagtgctgg gattacaggc gtgagccacc gtgcccggcc ccatctccct tctttttaca  10680 

gcaaggtgca tgttgcactg acttaccctt tattcctctt gtagtcactg gagctgtgtt  10740 

atttatttac tttattaatt tatttattta cttgaaacag agtctccctc tgttgcccag  10800 

gctggagtgc agtggcacaa tcctggctca ctgcaatctg gacctcccga gctcaagtga  10860 

tcctcccagc aggtgctatt gtaactgaag ccatatcaat aacagctcct tcaaaaccca  10920 

gctctgttgt ccttgatagg gttgccaatg caagtagctt atccacattc agagtattac  10980 

aaacttgtaa acttacacat tacttaacta tcactgattt ctctccttgg ttctatctga  11040 

aatggtttag ggaatcgttg gcagtatctg ttctttcaaa gccaattatt aatcagggct  11100 

tcattagaca gcattcacac atttgttttc ctaacatctg ttccattaat tttctaagaa  11160 

ccagcgtcag gcccaccaga tggcaatttc cagaaacact cactcatcct ttcctgaaga  11220 

tcagtagcgc atttgcttgt ttccaggcct ctgatccttc ctgccttgtc tgtgacttcc  11280 

tcaacaaccc ctcggggtga ttataagctc attccagcca tgtccattat gtgtggaatc  11340 

tgggtctatg agcttgaatt tcaacttcgg gctattgtta ccatttgtgc agaaagtttt  11400 

tctctgggtg ttaatactgc tcagaccttg aggctgtcaa gtgtacagga gcagagaaaa  11460 

gacaggctct ttctctttct ctggctcaga ggggtgggaa agagcattcg ctgcccacat  11520 

cttgtggaca gggatgaaga ggccagcagg tgacagcgtc tggcatagca cgtgctgtca  11580 

aggaaagaga aaggagccaa tggtgacaca ccagcttggt cagaggaagc atctgtgttt  11640 

ctgccaggct catgatgtgg gctctttgct atataagccc tgctttattg ggtctaaaac  11700 

acaaggttga gatgtcactg cactgctcaa acactttcag tgactcccta ctgttaatgg  11760 

gttaaaattt aaccttttag tctgacaggt ggaccccaat ctatcatctt gccctttctc  11820 

acctggctcc cacctgctgt ttcgggcccc tcttactcat ttgcatttcc tcccctccct  11880 

gcctttgctt ccagcagtca ctccatgtaa catttctttc ctccccatcc tcaaatcctc  11940 

tcaattttca ccccttccgt gaagtgctcc ttgccatttc cctcctttga tttcctgcag  12000 

caactcctgg acttctctga aaaccactgg tttcctgtcg ctcccctcac ctgtgctcct  12060 

gcattgtgac atcttccggg gcactctgtc ctattatttc tctctagtcc tgttatttgg  12120 

gcccatgtat taataccccc ccttagatat taacccataa gcctgaggct gcactttttt  12180 

gaattttgaa atcagacctt ggccttgacc ttgagcagca ggatataaat aactcttaca  12240 

tgcttagcgt tccaataatg gaacaccagg cataaatggg ttttaatccc cttgaaggca  12300 

ggggttgtgt ctactcatgt tttgcttccc aaggttagca ctatgcttgg catatagtag  12360 

ctgctcaata catctttgat aaatgaatga atgcccagat gaacaaacac acgaataaat  12420 

caactagctg taagatatgt aaactactag gtgctgatat ctttctagaa tcagtatttt  12480 

ctcaaaaagt aggaaaaacg ggttggaaaa cttaccagaa ctgagatgtc aaggcagtgg  12540 

gaggaggggg caattagatt tgactggcca gtctagtgcc atgttgtgga gctctgaggc  12600 

cacactgctc cttgctcagg actgtgtgtg attctagggc caccaagaat cttcctcgta  12660 

tctccacctt gcggtctgag gcctcaagcc tctagggagg tggcaggcgg gacggtggcc  12720 

acttggtgcc tgtccgttgg cagcacactg ttcctgcatg tctcgctcat gctgtgccct  12780 

ctgctctgct ttatctccta gacgaactgg ctgtctgccc taaaggaatt acctccaagg  12840 

ttttccgctt caatgtgtcc tcagtggaga aaaatagaac caacctattc cgagcagaat  12900 

tccgggtctt gcgggtgccc aaccccagct ctaagcggaa tgagcagagg atcgagctct  12960 

tccaggtaac tcctctctca gagcagaaac cacaccgacg ggaaagctgg ttcctttgcc  13020 

atatcagggc accactgggt gcagcgtttg gcagacctgg gtttgaatcc tggcttctct  13080 

gagccttcgt ttccgtatct gtgtctgtca ttaaaacact taagagttag ctaaggtgct  13140 

cgagggccat ggcattcagg aaccactggt ttcctatcgc tccctcacct gtgctctgca  13200 

ttgtgacatc ttctggggca ctctgtcctg gtctcgggta ctcactcctt tctctgccct  13260 

gtagatcctt cggccagatg agcacattgc caaacagcgc tatatcggtg gcaagaatct  13320 

gcccacacgg ggcactgccg agtggctgtc ctttgatgtc actgacactg tgcgtgagtg  13380 

gctgttgaga agaggtaggt ggacccttca gataagcatt tcagaatgaa cctcaggtcc  13440 

cttagtcctc catgaaatgg agggaagagg acagaattaa gggagtcaga gatctgggtt  13500 

caaaccctag ctttgccact gagtatcctc cattcattca ctcaactaat gtttattaaa  13560 

tgctcactgt aagacaggcc ctggggatgc agccacaggg ataggaacta tgagaaatag  13620 

aaagagggca atgtgacaat gagtgggtgg agtccaacag ggaaggtctc tatgatgaag  13680 

aaattcatgc attgacatct gaatgataag gatttagccc atgaagatca gaataaggga  13740 

tgtgctaagc aaaggcaaca gggaggccca ggccctcaag tggaaataag cttgatttgt  13800 

tctagcagca gcagcaaaca gatcggtgtg gctggagcat ggtgagctgg ggaggggaag  13860 

aggaggggag gtggtcaggg aggttgctgg ggccatataa tttattatta ctattattat  13920 

tattattatt attattatta ttattattat tattatttct tgagactgag tctcgctctg  13980 

ttgcctaagc tggagtgcag tggtgcgatc tctgctcact gcacctccac catctgggtt  14040 

caagcgattc tcacgtctca gtctcctgaa tagctgggac tacaggtgca cgctgccaca  14100 

cctggctaat ttttttgtat ttttagtaaa gatggggttt caccatgttg gccaggctgg  14160 

tctcgaactc ctgacctcaa gtgatccgcc cactttggct tccaaaagtg ctgggattat  14220 

agatgtgagc cagcatgccc agccaattta atttagaaca tcatcaggtc atggcgaggt  14280 

ttcaggactt attccaggtg tgatgagaag tgtgggagtg ctataaccag agctggggat  14340 

actcaagata cccaggaatt ccttcctgtc cctctactgg gtgtgaagtc aagagcctag  14400 

gagaacccac gtggatctgc caacggcagc tctgttggga attctgactc agacagctac  14460 

agggaggagg ggctgggtga ggtgatttga ttgacatctt taaaagatcc ctctagtttc  14520 

caggaggtca agggaagagg caggaaaatg agttaggagc cacggcggca gcccagatga  14580 

gagcaatatc taggccgaga ctagggcggt agcagtggat atgacgatca gatggatttg  14640 

ttctgtattt tgaaggtagc cagtagcaca ggctgattag gtatgggatg tgaggacaca  14700 

agagcattcc agataacttc taaatttttt cgacatccag gtggtatcac ttattgaaat  14760 

agggggccta ggagaagaac aggttctgtt ctgcccagtt aagtttaaaa ggccgggtcg  14820 

tcatgcaagt agagctatcc tggaggcata tcaaacttca catgtcccaa atatcttacc  14880 

cacacactgc cttcacctgg aaaatcaggc aatggttcct cctctatgta tgcctcacag  14940 

agctgtttta aggatcaaat gtatttgaga gaacttcatg gtttttacca tgttttacaa  15000 

gagtaagctt ttcttatttt agataaggaa acaggccgag agaagttaag tgacttgacc  15060 

gaggtcgtcc agtctggatt agaactttgg tgtctcatga caccatcctc tgtgtttctt  15120 

tcccttttct tggctggtac tgcctggtct gatgctcagt gggttggggt cacagatggc  15180 

agtcccatct gttccttctc ttcctcttgg gcaaggtttt ctcctgtcat cagctgctat  15240 

aaagccacag accatccaca tattgatgcc cagagtccct gaggcaggtg gatccttcta  15300 

agtccttggt gttttaggca actaagagtt aaagctcaaa ggcagagctt acaaactatt  15360 

tggcctgcac attgttttcg atttgactag tcatcaacag ttttgggtgt tttaagtcag  15420 

gagatttcac ataaaacctg gacttcttgc tgtttttgaa aaatcgaaag atttcctgat  15480 

aaccatccgt cagagatgag taccggccac cccttgagat ggggtatgca tcctcagtcc  15540 

ccgcagcccc caccattccc tattgttccc caagagtgag gctggcagtt cctgtttagc  15600 

atcatgtgct ggctcgattt tctcacttac gtgacctgcc tgggctcggt agccatcgag  15660 

tttgcattgc ttgatgccac ggagcccagg caagtcacat aaatgagaaa attccccatc  15720 

caaggggaat tatccagtgg ctgagttggc agatggtggg ctgaagatcc tgccccagtc  15780 

ccaagggtac ccaggaattt catcctgtcc ctcctactga gcatgaaggc aagagcctag  15840 

gagaacccac gtggatctgc cgacagcagc tctgttggga attctgactc agacggctac  15900 

agaaaggagg ggctggggaa attaatctct taacttccct gtctgatttt gatcagctcc  15960 

acctggattt ctttgaagcc ctgggcactt gaaggagttc ttattttcac agctgcagaa  16020 

ctcaatgaga agtttgcatt gagaatgatt tccatcctcc tgagacatca aaaagataat  16080 

ttcgtaataa aacctatggg tcccccaccc accaccccca cagctgccaa ttctgaggtt  16140 

agttcttcat tggaaccttc agttcacact taggcagata ctgcccacct ttcccacagg  16200 

ggaatcatca cacaggtttg tacttacaga gagcccatga gtctcttcag agttcatggc  16260 

ttcaaaccag gcaacaaagg actccaaatt ggagcagttg gcggtgtaga ttgatgagaa  16320 

aactgagtca cacatgtggt cttggattca ttcagcaaac atctgagagc ccttgtgcac  16380 

caggtaccca ggctgcattc tgggtcgcag agtgggttgg agtccacaag ggccgctggg  16440 

gacccaggtc tcataccctt gtgctattgc tcatatagtc tgcagcacct agctaggcag  16500 

ggccaggcct cccctagtag gctggggagg gagcctctca gcatcatggc tcaggaaagg  16560 

tgggacactg ggaaacaacc atcttgcatg ttggtgaatg tgtgggcatc ttccttggtg  16620 

gtctctgttg ccccctcttt acccctcctt gtctctaaca agagtttggg atctggcatc  16680 

aagctgtcag tgcttgaatc ctggcactac tatccactgg ttggatatct tgggcaagct  16740 

gtttaatctc cctaagcctc agtgttctta tctgtacaat gggggataat agttcttgct  16800 

ctatgggatt attatagaga tagaataagt tagtgattgt tgtcatcatc tctaagtcct  16860 

ctatctggat acagcctgtt ggccaagcca ggccaccctc ttgtgcttac atcctggacc  16920 

ctcactctcc ccaacacagc tgtctgcacc ttgactttct aactcacaga atcatggaac  16980 

tcactcactc caggtctctt ccagcctgcc ttgaaccagc ttgaccagga ggtacactgt  17040 

gttgttaggt agcccatttc agctactttg gctcctcctt ccagctgcct taacccaggg  17100 

agagggcaat cctcatacct ccctctccat tcacttctcc tccagctggg tctcagctgc  17160 

cttgttttat tggtctgcct tccctcactg agcgaacctg ctggaacaga gatcttcaag  17220 

ctcagcaggc gcagtgtgcc tcagaggttg ccctgactta gggtagagca gatctggtta  17280 

ggctctggag tttatggaag aaaggagctg ggttttaacc agtaggactg ggatgtccaa  17340 

gccaacctaa tgttgatggg aatctccagt cttgtaggct tgtatccttc tgtctatgag  17400 

aaggttgctg ggccaggttg gactttttgg ggtttgccct gatggcacaa tttcaggaag  17460 

actccaggtt gtctccaagg ccagatgctc acacacatgg tgtgtggcct cagggagcag  17520 

caaatcaatc aactcatcga gaggctgacc ctgaatccag agaggtccat cccccatgac  17580 

ctctcctaca cctggccaca actggctgct atcccaggat caacagagct tcccaaatta  17640 

agtcttaccc accagaattg acatgatacc agatcctacc tggttttctg tgtaaaacta  17700 

cttgtctcag tgcagacttt tcattttctt tctttttcct tcacttcact caacctcatt  17760 

tactgagttc ctgcttcctt gattttggtt gccaaaacat gtgatcccac aactactagg  17820 

cgaatgggct ggagttgccc ttcacgcacc agaatgtcag gcaccaaacc ctggcacctc  17880 

caggcttcat catgctcact gtctccgtgc tcaagtcatc tggaccctca gccatagttc  17940 

tctatcatct cctctgtgct gttttctaag ttcacaactg tcttcccaag ctgttcatct  18000 

tggtgagcaa tccacattcc aacttcccat catggtgaac aaggattccc aaagaagaaa  18060 

tgtccaagta atttttacct cgttgtgtcc ttgttcctta ggaaaaatat ggcccagtgt  18120 

aaccgtgatc ttcatcttat ctcttctaga atctgccact ttgtctaggc ccacagtcat  18180 

ggccctggta gaaatgcttg catcttccac tgagacggat tctgcttcct gtttgcaagt  18240 

gaccttgcct ctagcattgg ttcctgactt gaatcttttt gccagcatta catcctgtgg  18300 

ctttttgaat ttgtggcctt ttgaatcaca gtctctgtgc tcggagactc tttagttact  18360 

ccctgtccgt agtctcttcc agtccctctg cctcttgccc tgatccttct gcagtagatt  18420 

ggctcaacga tcccctctcc caaaaaccat gtagatggca tgagtttttg ctttccttac  18480 

tgtatctgtg tacttttgcc tccaagtagg taggtgacaa ttttctctta ataccatcct  18540 

ttcaaaggga atgattattc cacttctgtt tcatgatggc cactagtgta tgcccccatt  18600 

tggtgattca taatacaaca ggaatacctg gaatgtggca acgtgcgctt gaaggtccac  18660 

tctgagctct ggaggcactt tctgtctcct gtgaccctca atagaactca gttcctatga  18720 

gtccctatag gagcagaggc cctggctcac ccttgcatgg gtggtgggca cacttcccca  18780 

tcacacagag gtgcttttct cactaattct gtcttatttt gcagagtcca acttaggtct  18840 

agaaatcagc attcactgtc catgtcacac ctttcagccc aatggagata tcctggaaaa  18900 

cattcacgag gtgatggaaa tcaaattcaa aggtaacaaa atgaatgtgt atggtaggat  18960 

gggtgagtgg gggggaagtt aatgggacag gatagtgcag gagaccctta ccagacctca  19020 

agaaaagaaa ccaagctcct tcagagagat agccgactat tttgtacttg agtaattctg  19080 

ctttgcccca aaagcaatga gtgtaacttt cagtttatag cttagagaat gcattagcct  19140 

ctggaagaca acacgtactg gtaagtggtg aaactgtgta ggcaagccat ctagcctctg  19200 

gtcctagtga aatgggatga gagttcctct ttcacagtgt tgtcaggcaa ttaaatgaga  19260 

taatgaataa ggaacacagt gccagaaatg ctcaacagat aggacttgtc ttcctcctcc  19320 

ctaaaagaaa ttaagtttgg ctgtcctgaa catgagtgcc cagacaactg agctttccag  19380 

atgtgcaggg cctacctgac cttgcccagg ggtcctctgg gtttggatga ttgcttcgag  19440 

cctcagggtg tttgtccccg gggtgtttgg gtagagatgg cgaagtcgtt gagagtcatt  19500 

tctggttttc cattatgttt gcaaggaact cagccttgat gatctctgga gttcagggaa  19560 

gttctctttt cctttcatat tcccattttg ggtaactgcg gaacgcctga ggtcagaggc  19620 

ttgtctggga aaaggtgcag gcctcttttg gctcagcgct ggacagtgat cttaccccac  19680 

atgggctcta ttttacagcc ttttcttaaa gccaaagatt tgacactgta accacagaac  19740 

cttagagccc agaggacctt tggagtcctt acctacaggc cagcttagga tgaaaccttc  19800 

ccatttcagt gaatacttat cttgttcata aagattttca gaaaaaggga tttgtgatcc  19860 

aggtctctca tttatcttac agatctggtg ccccttcctg agacccgagc cactcctgta  19920 

gctatgtaaa ttaattcctc atgttcctgt cctcagtgag gatggacaac agttggcagc  19980 

tgtccttcgc aaaatcgatc tccatttagt tgagactctt tctaagtcag tctccagtct  20040 

tttccttttt tttcagatga aacgggttga tgaatttaga ttttctttcc agagcttatt  20100 

tgctatttat catatgcttc attatctcat ctgaatttta tagtgaaaac acttcaaaga  20160 

ctctaagtgc aatgtgaatg ttaacaatta tacagtcttg tttctttggc tcccattcag  20220 

gctatcttaa gttgtgacac cttaaatttt tggtaggact tctgcgttat cttggtccat  20280 

tcacatttta agaggaaact cacaccccaa gattctaagt ctagaatcta aagtgacaat  20340 

ccagggctgg gtgcagtggc tcacacctgt aatcttagca ctctgggagg ccgaggtggg  20400 

cggatcactt gaggtcagga gttcgagacc agcctggcca acatggtgaa accctgtctc  20460 

tactaaaaat acaaaaatag ctgggcatgg tggcatgcac ctgtaatccc agctacttgg  20520 

gaggctgagg caggagaatc tcttgaaccc cggaggcaga ggttgcagtg agccaagatg  20580 

gggccactgc actccagcct ggccaacaga gcgagactct gtctcaaaac aaacaaacaa  20640 

acaaataaac aagccgacaa cccggagata tgtgttaggt acccacttag taacagggat  20700 

gcttcatagg tccatgaaga ttcctaggaa tctcagcaag ggctttctgc cccttggaag  20760 

atttctatac aagggtatgg ggatctgaac acggggcatc tttcagtggg catccttaca  20820 

ataataatga gttcttttga tactggcttc tccatctgct cttccccttt ctgcacctgg  20880 

acatcagaat taagctgcac ttgtccccca cacctccctc gcagactgca ctgcccctcc  20940 

tcctgggcag tgatggggcg tgtggaggag gcagcctcca agggctctgc tctcttcaga  21000 

caggagattg tcactttcct tcccttcttc aggcgtggac aatgaggatg accatggccg  21060 

tggagatctg gggcgcctca agaagcagaa ggatcaccac aaccctcatc taatcctcat  21120 

gatgattccc ccacaccggc tcgacaaccc gggccagggg ggtcagagga agaagcgggc  21180 

tttggacacc aattactgct tccggtgaga ctgggcccac atgggaacca acatctactg  21240 

cctgcctact gcccaatggc taggtcaggc cccagagcca agccacactc aacagagggt  21300 

ccctgatgct attcacaaac atctccagga agaagactga aaatctctca cagagatttt  21360 

ctctgtgaaa tctctttctg ttttcctggg agtcccactg tttttccata ggctaactct  21420 

ggaaggagct ggctgaagta aatgaggaaa actctgtgag gaggagtgtt gctaaaatag  21480 

tttggattgg agaggcttgg tcaaagcctc tccatgattt ccatgtttta agcacttgta  21540 

gagtgtatgt gtgagattaa tgtaggagtt tccattaaag aagtgctcag ttagttccct  21600 

atgaagggtc caaggatgct actggatgga ggcaaataga atggtctcca tttgaacgga  21660 

aagttggagc tagagaaatt aacaaatgaa ttcagaaatt actgggtagc cacaatagag  21720 

agatagaaaa gacccagctt ctcttactca ggagcagttt cactgctcat ttataaaggg  21780 

aggataaagt atgcatgcca actactaaaa gggagaagat gatcacgccc atgagagggt  21840 

ccgaaggatt aagtgcttta actgggagca atgagttcac ctatggacac aggaaaggct  21900 

tcagaagggt gtggaatttg agcagaggct tcaaggatga ggtttgggga tgaagggatc  21960 

caggagggga agcactgcag gtaaaggaag gagcatgagt gagtgagtcc atctggcctc  22020 

agaggatgcc gaaatgaagc tggggagaac gaattgggaa atatttagat ctggaaacca  22080 

tgtcttatga ggatggttgg gcttgggaaa ctagggatgt tcagttgggg gaacagctca  22140 

acagcaaggt ataggagagg caaggtagtt gccacaaagg ctggcatgta gatttattta  22200 

ttccatcagt taggaacaaa tcagttagga acgaatcagt tatcagttag gaacgaatca  22260 

gttaggaacg aatgatcttg gccagtgact ttaatcatag tgatttcttg ttcacaagag  22320 

gcctgtagca gcagggttag ggtgggctgg ctgcttaatg atgccaccga agactcaggc  22380 

tttctctctt tccagtctgc catgcttagt atgttggctt ttcatccttt tccttatggt  22440 

cacaagtgat tgctagggct tcaggcacct tgtccacatt taagacaaga aggaatgagg  22500 

gaaggggaag agccagaagc tttcttcttc catctgtcac ttttataaat aatcgaatat  22560 

ctttgccaga acttgccctt accccctgca tacttctcct tgtctgatgg gtcattactg  22620 

ggtcacatga ccaccgctac tctcaaggga ggacaggaaa aggagcatct ggtgttttcg  22680 

actctgtaat gggatgtcac aagggagaag ggagttagga atggttattg aatagataac  22740 

caatggtgtc tgccatggtt cacccttgga acccacaaga agccagattt tggctaatag  22800 

ctgcctttgg aagtagcaag ttctaagtct cagcagatat taaagcagag gctagctgtc  22860 

cacttggcaa gcggtattat tgccaaatga tctctgtagg caacagaatt gaaggggcct  22920 

gataatcaca tttgggtgat ctgttgaacc acggttctaa tagaaggata tgccttattt  22980 

ggctaaatgg cctttggatt gagtctcagc agtcacctac tatagtagtc aagctgcata  23040 

aacttagaat tgatttctgt ctgggtgcac attaggaggg taaaaataaa accaacctca  23100 

acaaagctga gttggctaat aacattcagt gcttggtttt atgtggagcg cttcatagca  23160 

ctgctttgta ttgtgataga aaagggtgca gggccacctt tgcctcttta ctcttcctca  23220 

tgcagcattt atctctgttt cttacatcct tgggatcctg gtctttaaca catgaatgtg  23280 

cctttctggt ttcttctgcc ccatgccctt gaacctggga ttccattgat cagagccacc  23340 

tatgatggca tgaaaggact ccaagggaga atatgagagc attcatgaag tttcttttat  23400 

tggtggtttt aagttgattt tctataaatg ttttttgttg tagcaacttg gaggagaact  23460 

gctgtgtgcg ccccctctac attgacttcc gacaggatct gggctggaag tgggtccatg  23520 

aacctaaggg ctactatgcc aacttctgct caggcccttg cccatacctc cgcagtgcag  23580 

acacaaccca cagcacggta tggagcaggc tcatgccatc tgaggcactg gggctgaccg  23640 

accagaccac ttgttaaaaa gaatgagtga aggattgaat gttgagtgag caaatgatgg  23700 

tctggggtga gtaaaattcc tctggtgaag gttctgatct tggcaccctg actcagttct  23760 

actcagtcac attctgccct tttaaattcc taccatagtt tcaaccagct ttttaatttg  23820 

ctttgtccaa agctctctgt accgtagaaa ttaattaact gccacgtgtt accctatgtc  23880 

agagatatgt gcatgtggct ggcaaggaac actgaagtag aaaggcttct atcagaactc  23940 

tgatcattcc agcctctcga atgtagaaaa cttactctga aagacaacaa caaggatgga  24000 

caagaagcta atttgaaatg ctaggaacag aaagtgagat agccgagagt ccacaacccc  24060 

tgaaactagt ctgtctttcc cttgagggga atcaaaaata gggcagtaat ttgtgaagca  24120 

tgtttccttc tagctcattg tttctgcatc ctgtctgggg ccctgcacgc tgcacttact  24180 

aaatggctca aggcaatgtt ttgtgagaac atttctcacc gaagtgatca gttccagcta  24240 

ggaagagcaa tgtaagtgtt ttcttaaaag ccaagatagg ccgggcacgg tggctcacgc  24300 

ctgtaatccc agcactctgg gaggccaagg cgggtggatc acaaggtcag gagttcgaga  24360 

ccagcctggc caacatggtg aaaccctgtc tctaccaaaa atgtaaaaaa ttagccgggt  24420 

gtggtggcac gcgcttgtaa tcccagctac ttgggaggct gaggcaggag aatcgcttga  24480 

acccaggagg cggaagttgc agtgagccaa gattgcacca ctgcacttca gcctgggtga  24540 

aagagtgaga ctctgactca aaaaaaaaaa aaaaaaaaaa aaaaaaaagc caagataatt  24600 

cgttatcagt gtagtaactg tcatatgctt caactcatca ccccaaacaa gtgcagttgt  24660 

tctgcttctg tcattgtgga tgaacagcaa ctacacaact actcactcac ccaaaccagg  24720 

ttccaacagt tttttttttt ttcttttttt tgagacagac tcactctgtg gctcaggctg  24780 

gagtgcggtg gcactatttc tgcttactgc aacctctgct tcctggattc aggtgatcct  24840 

cccacctcag cctcccaagt agctgggatt acaggcgcct gccaccacag ccagctaatt  24900 

tttttgtatt tttagtagag atggggtttc accatgctgg ccaggctggt ctcaaactcc  24960 

tgacctgaag tgatctgcct gcctctgcct cccaagtcct gggactacag ctgtgagcca  25020 

ctgcacccgg cccccaacag ttcctcttaa agagcacccg tggccattct agcacttgct  25080 

cagcttctgg gactgcccct ggaactgggc gacttcctgg gttccaggcc tgagacttgg  25140 

ccttccaacc tctcactgac atgtcccttc ccaggtgctg ggactgtaca acactctgaa  25200 

ccctgaagca tctgcctcgc cttgctgcgt gccccaggac ctggagcccc tgaccatcct  25260 

gtactatgtt gggaggaccc ccaaagtgga gcagctctcc aacatggtgg tgaagtcttg  25320 

taaatgtagc tgagacccca cgtgcgacag agagagggga gagagaacca ccactgcctg  25380 

actgcccgct cctcgggaaa cacacaagca acaaacctca ctgagaggcc tggagcccac  25440 

aaccttcggc tccgggcaaa tggctgagat ggaggtttcc ttttggaaca tttctttctt  25500 

gctggctctg agaatcacgg tggtaaagaa agtgtgggtt tggttagagg aaggctgaac  25560 

tcttcagaac acacagactt tctgtgacgc agacagaggg gatggggata gaggaaaggg  25620 

atggtaagtt gagatgttgt gtggcaatgg gatttgggct accctaaagg gagaaggaag  25680 

ggcagagaat ggctgggtca gggccagact ggaagacact tcagatctga ggttggattt  25740 

gctcattgct gtaccacatc tgctctaggg aatctggatt atgttataca aggcaagcat  25800 

tttttttttt tttttaaaga caggttacga agacaaagtc ccagaattgt atctcatact  25860 

gtctgggatt aagggcaaat ctattacttt tgcaaactgt cctctacatc aattaacatc  25920 

gtgggtcact acagggagaa aatccaggtc atgcagttcc tggcccatca actgtattgg  25980 

gccttttgga tatgctgaac gcagaagaaa gggtggaaat caaccctctc ctgtctgccc  26040 

tctgggtccc tcctctcacc tctccctcga tcatatttcc ccttggacac ttggttagac  26100 

gccttccagg tcaggatgca catttctgga ttgtggttcc atgcagcctt ggggcattat  26160 

gggttcttcc cccacttccc ctccaagacc ctgtgttcat ttggtgttcc tggaagcagg  26220 

tgctacaaca tgtgaggcat tcggggaagc tgcacatgtg ccacacagtg acttggcccc  26280 

agacgcatag actgaggtat aaagacaagt atgaatatta ctctcaaaat ctttgtataa  26340 

ataaatattt ttggggcatc ctggatgatt tcatcttctg gaatattgtt tctagaacag  26400 

taaaagcctt attctaaggt gtatgtctga ctcgataaat atccttcaat tacccttaat  26460 

tccatgtccg catctactaa tcagaggtaa ccagaatctg gggcagagaa tctgtcaatc  26520 

accaaacaca ttgcttagcg caactcccct tacacagggg gcacaccgtg ctgacttccg  26580 

cctgctaaga agactgcact attgcttgtg cgtctttcct tcttgcagag tatattattc  26640 

tcagagacac agaggcagtg gctcagattg gcagaaagca catacgaatt tgacctccag  26700 

atacctgtgg gcaggatccc ctggtgtgaa tccttgcata tggaaacatg gtttatttac  26760 

taactataaa ttaccaacat cactgttttc gaaaatctcc ccccacccgt atcagttgag  26820 

aatgagagaa aatgtgtatg gcaaatggcc taaaaatatg aggctaattc tgttctcagg  26880 

ctaagcctaa aagagctaac caggaacccc ttttcagatt gtggccctct tgtcagggat  26940 

ctgggaacca tagctctctt ttgagtgagg ccgtggatcc cactgtggta tggacatcca  27000 

cgtggggcag ccgtccacag tgagcgggca cagagcgaca ggccatctag cagctcctga  27060 

gaaacacatt cttctagtga gatcactttg cctctagtaa aagaaaagtc tatctgaagc  27120 

taaagtatgc aggctgaagg atgtgtctga tgtgttcatg cctgtgtgtg tgtgtacgta  27180 

tgtgtgtccc ttgttccttg agttctccaa gactgaagtg agtttggtca gtactttttt  27240 

ccccgctttg tccacgccca gccaattcta agggtttccc tctcagtctt catcccatgt  27300 

ggcacccact gacttccacc atcgttcgga gggctatgtc cctgtcctaa aatcctggct  27360 

ggcgggatta tactccatct tgctccaggg agcccggggg cacagagagg ggatgcagta  27420 

agctcagcac atcagccacg ggctgctgct gtcccagcat cagtctattc acctgagggc  27480 

cattctcaaa ttcactgggc atgataaccc ccaagggttt atttaaaatg tacagtttca  27540 

ggctgcagct ctagaggtac ctttaggact gcagtagggt ccaggttgcc tcaaaactat  27600 

taacacattt tgagaagtgg tgcattacac ccgacatcac tcgtccttac ttgaggcttc  27660 

aatatagaaa agggaaataa tttttttggc cccagatgaa accccctttt atccaacttc  27720 

acagcacatt gctaaaacat tggcctgtgg ttcatcctaa tgaaccaagg cagctagata  27780 

catttcctgg attattccaa agaaaacaga acgtggtgca attccaaatg gtctttttct  27840 

tcagagctct acttgtaggt taggcagcag acacaaatac ggacaggggt ctgaagtcag  27900 

cccccctatt ctacacatca ctgacaggtg ctacagaaac ttgaatcact gtcttcaaaa  27960 

atcgacgctt gttttggggg agggtaaaga gtgacattgg aaatatcacc tctgttggag  28020 

gagggctcat agcatctgtc tggtttattg ccaagtgaag tccagtccca ataaaaatga  28080 

cttgaaagca cttggacaca tgaagggaga gatctggtgt cttagcacct gatcagcata  28140 

tggtaagtgc agtaagaaat tagctggaag acttcattct gattggttac ctagggcaat  28200 

ttcaagcatt atagtctaaa acttctttta ctgggctcat acttttttcc cttgtcacaa  28260 

gattcacgtg gtgagtcctc ccaaacccct tatttcccaa ttctacacct catcagggga  28320 

ctatggagga attctaattt gtcccctaat caactcacaa ccaattgagc aaatagttaa  28380 

ggtggtcctg aaactactct ccagtcccgg ggacatttgg aatgcattct tacttctctt  28440 

ctgaaaccca tagacctcat ccttcacaag caaacaaaat gtgtccatgt gcccaaacct  28500 

ttgttttcat tcagtaagaa ggcaataaag tcccttttct gcccttttag gtgtcaattt  28560 

tttctttttt tttttttttt tttttttttt caggtggagt cttgctctgt tgcccaggct  28620 

ggagtgcagt agcatgatct cggctcactg caacctctgc ctcccaggtt caagcaattc  28680 

tcctgcctca gcctcccgag tagctgagat tacaggcgcc tgccactgcg cccagctaat  28740 

ctttgtattt ttagtagaga cgaggtttca ccatctttgc caggctagtc ttgaactcct  28800 

gacctcgtga ttcacccgcc tcgacctccc aaagtgctgg gattacaggc gtgagccact  28860 

gcgctcagcc aggtgtcaat tttctttttg gatttcaaca ctgagtccat agtaccctgc  28920 

tgaagaagcc ccagagcctg ggttctcccc tgataactct ctagggcagc taagttaatc  28980 

cttcagtgga ctctgctgtc                                              29000 

 
           
             467  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            467 

ttgttgtcca tgtgtctaaa                                                 20 

 
           
             468  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            468 

ttcaggactt ccaggaagcg                                                 20 

 
           
             469  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            469 

aggtgcatga actcactgca                                                 20 

 
           
             470  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            470 

cggcaaggcc tggagaggaa                                                 20 

 
           
             471  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            471 

aagtgcatct tcatgtgtga                                                 20 

 
           
             472  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            472 

tttgcaagtg catcttcatg                                                 20 

 
           
             473  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            473 

agccctttgc aagtgcatct                                                 20 

 
           
             474  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            474 

ccagagccc tttgcaagtg                                                  20 

 
           
             475  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            475 

aagttcagca gggccaggac                                                 20 

 
           
             476  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            476 

aagtggacag agagaggctg                                                 20 

 
           
             477  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            477 

tgcaagtgga cagagagagg                                                 20 

 
           
             478  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            478 

gtggtgcaag tggacagaga                                                 20 

 
           
             479  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            479 

ttgatgtggc cgaagtccaa                                                 20 

 
           
             480  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            480 

tcttcttgat gtggccgaag                                                 20 

 
           
             481  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            481 

cctcttcttc ttgatgtggc                                                 20 

 
           
             482  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            482 

tccaccctct tcttcttgat                                                 20 

 
           
             483  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            483 

tggcttccac cctcttcttc                                                 20 

 
           
             484  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            484 

cctaatggct tccaccctct                                                 20 

 
           
             485  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            485 

tgtcccctaa tggcttccac                                                 20 

 
           
             486  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            486 

agatctgtcc cctaatggct                                                 20 

 
           
             487  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            487 

ctcaagatct gtcccctaat                                                 20 

 
           
             488  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            488 

ttgctcaaga tctgtcccct                                                 20 

 
           
             489  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            489 

ggacgtgggt catcaccgtt                                                 20 

 
           
             490  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            490 

atcatgtcga atttatggat                                                 20 

 
           
             491  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            491 

ccctggatca tgtcgaattt                                                 20 

 
           
             492  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            492 

tccactgagg acacattgaa                                                 20 

 
           
             493  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            493 

ttctccactg aggacacatt                                                 20 

 
           
             494  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            494 

atttttctcc actgaggaca                                                 20 

 
           
             495  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            495 

tgggcacccg caagacccgg                                                 20 

 
           
             496  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            496 

gtgggcagat tcttgccacc                                                 20 

 
           
             497  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            497 

cgcacagtgt cagtgacatc                                                 20 

 
           
             498  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            498 

aaggtgtgac atggacagtg                                                 20 

 
           
             499  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            499 

gctgaaaggt gtgacatgga                                                 20 

 
           
             500  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            500 

attgggctga aaggtgtgac                                                 20 

 
           
             501  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            501 

tctccattgg gctgaaaggt                                                 20 

 
           
             502  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            502 

aatttgattt ccatcacctc                                                 20 

 
           
             503  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            503 

tctccacggc catggtcatc                                                 20 

 
           
             504  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            504 

ttgcggaagc agtaattggt                                                 20 

 
           
             505  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            505 

tgagcagaag ttggcatagt                                                 20 

 
           
             506  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            506 

gggcctgagc agaagttggc                                                 20 

 
           
             507  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            507 

ggcaagggcc tgagcagaag                                                 20 

 
           
             508  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            508 

gcactgcgga ggtatgggca                                                 20 

 
           
             509  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            509 

tcagggttca gagtgttgta                                                 20 

 
           
             510  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            510 

gacttcacca ccatgttgga                                                 20 

 
           
             511  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            511 

gggtctcagc tacatttaca                                                 20 

 
           
             512  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            512 

agtgaggttt gttgcttgtg                                                 20 

 
           
             513  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            513 

gaaacctcca tctcagccat                                                 20 

 
           
             514  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            514 

agagttcagc cttcctctaa                                                 20 

 
           
             515  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            515 

ttagggtagc ccaaatccca                                                 20 

 
           
             516  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            516 

agccattctc tgcccttcct                                                 20 

 
           
             517  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            517 

tcagatctga agtgtcttcc                                                 20 

 
           
             518  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            518 

tccagattcc ctagagcaga                                                 20 

 
           
             519  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            519 

gtataacata atccagattc                                                 20 

 
           
             520  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            520 

aaaatgcttg ccttgtataa                                                 20 

 
           
             521  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            521 

ctgggacttt gtcttcgtaa                                                 20 

 
           
             522  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            522 

ttgcaaaagt aatagatttg                                                 20 

 
           
             523  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            523 

ttaattgatg tagaggacag                                                 20 

 
           
             524  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            524 

ctggattttc tccctgtagt                                                 20 

 
           
             525  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            525 

aactgcatga cctggatttt                                                 20 

 
           
             526  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            526 

atacagttga tgggccagga                                                 20 

 
           
             527  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            527 

atccaaaagg cccaatacag                                                 20 

 
           
             528  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            528 

ccaccctttc ttctgcgttc                                                 20 

 
           
             529  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            529 

gtctaaccaa gtgtccaagg                                                 20 

 
           
             530  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            530 

tgcatggaac cacaatccag                                                 20 

 
           
             531  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            531 

atgccccaag gctgcatgga                                                 20 

 
           
             532  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            532 

aatgaacaca gggtcttgga                                                 20 

 
           
             533  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            533 

cacctgcttc caggaacacc                                                 20 

 
           
             534  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            534 

tgtagcacct gcttccagga                                                 20 

 
           
             535  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            535 

agtcactgtg tggcacatgt                                                 20 

 
           
             536  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            536 

agtaatattc atacttgtct                                                 20 

 
           
             537  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            537 

atatttattt atacaaagat                                                 20 

 
           
             538  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            538 

ctgttctaga aacaatattc                                                 20 

 
           
             539  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            539 

ctgctggaag caaaggcagg                                                 20 

 
           
             540  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            540 

gaggagttac ctggaagagc                                                 20 

 
           
             541  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            541 

gtccacctac ctcttctcaa                                                 20 

 
           
             542  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            542 

atgccatcta catggttttt                                                 20 

 
           
             543  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            543 

ttgtccacgc ctgaagaagg                                                 20 

 
           
             544  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            544 

ccagtctcac cggaagcagt                                                 20 

 
           
             545  
             14121  
             DNA  
             Homo sapiens  
             
               CDS  
               (129)...(13820)  
             
           
            545 

attcccaccg ggacctgcgg ggctgagtgc ccttctcggt tgctgccgct gaggagcccg     60 

cccagccagc cagggccgcg aggccgaggc caggccgcag cccaggagcc gccccaccgc    120 

agctggcg atg gac ccg ccg agg ccc gcg ctg ctg gcg ctg ctg gcg ctg     170 
         Met Asp Pro Pro Arg Pro Ala Leu Leu Ala Leu Leu Ala Leu 
          1               5                   10 

cct gcg ctg ctg ctg ctg ctg ctg gcg ggc gcc agg gcc gaa gag gaa      218 
Pro Ala Leu Leu Leu Leu Leu Leu Ala Gly Ala Arg Ala Glu Glu Glu 
 15                  20                  25                  30 

atg ctg gaa aat gtc agc ctg gtc tgt cca aaa gat gcg acc cga ttc      266 
Met Leu Glu Asn Val Ser Leu Val Cys Pro Lys Asp Ala Thr Arg Phe 
                 35                  40                  45 

aag cac ctc cgg aag tac aca tac aac tat gag gct gag agt tcc agt      314 
Lys His Leu Arg Lys Tyr Thr Tyr Asn Tyr Glu Ala Glu Ser Ser Ser 
             50                  55                  60 

gga gtc cct ggg act gct gat tca aga agt gcc acc agg atc aac tgc      362 
Gly Val Pro Gly Thr Ala Asp Ser Arg Ser Ala Thr Arg Ile Asn Cys 
         65                  70                  75 

aag gtt gag ctg gag gtt ccc cag ctc tgc agc ttc atc ctg aag acc      410 
Lys Val Glu Leu Glu Val Pro Gln Leu Cys Ser Phe Ile Leu Lys Thr 
     80                  85                  90 

agc cag tgc acc ctg aaa gag gtg tat ggc ttc aac cct gag ggc aaa      458 
Ser Gln Cys Thr Leu Lys Glu Val Tyr Gly Phe Asn Pro Glu Gly Lys 
 95                 100                 105                 110 

gcc ttg ctg aag aaa acc aag aac tct gag gag ttt gct gca gcc atg      506 
Ala Leu Leu Lys Lys Thr Lys Asn Ser Glu Glu Phe Ala Ala Ala Met 
                115                 120                 125 

tcc agg tat gag ctc aag ctg gcc att cca gaa ggg aag cag gtt ttc      554 
Ser Arg Tyr Glu Leu Lys Leu Ala Ile Pro Glu Gly Lys Gln Val Phe 
            130                 135                 140 

ctt tac ccg gag aaa gat gaa cct act tac atc ctg aac atc aag agg      602 
Leu Tyr Pro Glu Lys Asp Glu Pro Thr Tyr Ile Leu Asn Ile Lys Arg 
        145                 150                 155 

ggc atc att tct gcc ctc ctg gtt ccc cca gag aca gaa gaa gcc aag      650 
Gly Ile Ile Ser Ala Leu Leu Val Pro Pro Glu Thr Glu Glu Ala Lys 
    160                 165                 170 

caa gtg ttg ttt ctg gat acc gtg tat gga aac tgc tcc act cac ttt      698 
Gln Val Leu Phe Leu Asp Thr Val Tyr Gly Asn Cys Ser Thr His Phe 
175                 180                 185                 190 

acc gtc aag acg agg aag ggc aat gtg gca aca gaa ata tcc act gaa      746 
Thr Val Lys Thr Arg Lys Gly Asn Val Ala Thr Glu Ile Ser Thr Glu 
                195                 200                 205 

aga gac ctg ggg cag tgt gat cgc ttc aag ccc atc cgc aca ggc atc      794 
Arg Asp Leu Gly Gln Cys Asp Arg Phe Lys Pro Ile Arg Thr Gly Ile 
            210                 215                 220 

agc cca ctt gct ctc atc aaa ggc atg acc cgc ccc ttg tca act ctg      842 
Ser Pro Leu Ala Leu Ile Lys Gly Met Thr Arg Pro Leu Ser Thr Leu 
        225                 230                 235 

atc agc agc agc cag tcc tgt cag tac aca ctg gac gct aag agg aag      890 
Ile Ser Ser Ser Gln Ser Cys Gln Tyr Thr Leu Asp Ala Lys Arg Lys 
    240                 245                 250 

cat gtg gca gaa gcc atc tgc aag gag caa cac ctc ttc ctg cct ttc      938 
His Val Ala Glu Ala Ile Cys Lys Glu Gln His Leu Phe Leu Pro Phe 
255                 260                 265                 270 

tcc tac aac aat aag tat ggg atg gta gca caa gtg aca cag act ttg      986 
Ser Tyr Asn Asn Lys Tyr Gly Met Val Ala Gln Val Thr Gln Thr Leu 
                275                 280                 285 

aaa ctt gaa gac aca cca aag atc aac agc cgc ttc ttt ggt gaa ggt     1034 
Lys Leu Glu Asp Thr Pro Lys Ile Asn Ser Arg Phe Phe Gly Glu Gly 
            290                 295                 300 

act aag aag atg ggc ctc gca ttt gag agc acc aaa tcc aca tca cct     1082 
Thr Lys Lys Met Gly Leu Ala Phe Glu Ser Thr Lys Ser Thr Ser Pro 
        305                 310                 315 

cca aag cag gcc gaa gct gtt ttg aag act ctc cag gaa ctg aaa aaa     1130 
Pro Lys Gln Ala Glu Ala Val Leu Lys Thr Leu Gln Glu Leu Lys Lys 
    320                 325                 330 

cta acc atc tct gag caa aat atc cag aga gct aat ctc ttc aat aag     1178 
Leu Thr Ile Ser Glu Gln Asn Ile Gln Arg Ala Asn Leu Phe Asn Lys 
335                 340                 345                 350 

ctg gtt act gag ctg aga ggc ctc agt gat gaa gca gtc aca tct ctc     1226 
Leu Val Thr Glu Leu Arg Gly Leu Ser Asp Glu Ala Val Thr Ser Leu 
                355                 360                 365 

ttg cca cag ctg att gag gtg tcc agc ccc atc act tta caa gcc ttg     1274 
Leu Pro Gln Leu Ile Glu Val Ser Ser Pro Ile Thr Leu Gln Ala Leu 
            370                 375                 380 

gtt cag tgt gga cag cct cag tgc tcc act cac atc ctc cag tgg ctg     1322 
Val Gln Cys Gly Gln Pro Gln Cys Ser Thr His Ile Leu Gln Trp Leu 
        385                 390                 395 

aaa cgt gtg cat gcc aac ccc ctt ctg ata gat gtg gtc acc tac ctg     1370 
Lys Arg Val His Ala Asn Pro Leu Leu Ile Asp Val Val Thr Tyr Leu 
    400                 405                 410 

gtg gcc ctg atc ccc gag ccc tca gca cag cag ctg cga gag atc ttc     1418 
Val Ala Leu Ile Pro Glu Pro Ser Ala Gln Gln Leu Arg Glu Ile Phe 
415                 420                 425                 430 

aac atg gcg agg gat cag cgc agc cga gcc acc ttg tat gcg ctg agc     1466 
Asn Met Ala Arg Asp Gln Arg Ser Arg Ala Thr Leu Tyr Ala Leu Ser 
                435                 440                 445 

cac gcg gtc aac aac tat cat aag aca aac cct aca ggg acc cag gag     1514 
His Ala Val Asn Asn Tyr His Lys Thr Asn Pro Thr Gly Thr Gln Glu 
            450                 455                 460 

ctg ctg gac att gct aat tac ctg atg gaa cag att caa gat gac tgc     1562 
Leu Leu Asp Ile Ala Asn Tyr Leu Met Glu Gln Ile Gln Asp Asp Cys 
        465                 470                 475 

act ggg gat gaa gat tac acc tat ttg att ctg cgg gtc att gga aat     1610 
Thr Gly Asp Glu Asp Tyr Thr Tyr Leu Ile Leu Arg Val Ile Gly Asn 
    480                 485                 490 

atg ggc caa acc atg gag cag tta act cca gaa ctc aag tct tca atc     1658 
Met Gly Gln Thr Met Glu Gln Leu Thr Pro Glu Leu Lys Ser Ser Ile 
495                 500                 505                 510 

ctc aaa tgt gtc caa agt aca aag cca tca ctg atg atc cag aaa gct     1706 
Leu Lys Cys Val Gln Ser Thr Lys Pro Ser Leu Met Ile Gln Lys Ala 
                515                 520                 525 

gcc atc cag gct ctg cgg aaa atg gag cct aaa gac aag gac cag gag     1754 
Ala Ile Gln Ala Leu Arg Lys Met Glu Pro Lys Asp Lys Asp Gln Glu 
            530                 535                 540 

gtt ctt ctt cag act ttc ctt gat gat gct tct ccg gga gat aag cga     1802 
Val Leu Leu Gln Thr Phe Leu Asp Asp Ala Ser Pro Gly Asp Lys Arg 
        545                 550                 555 

ctg gct gcc tat ctt atg ttg atg agg agt cct tca cag gca gat att     1850 
Leu Ala Ala Tyr Leu Met Leu Met Arg Ser Pro Ser Gln Ala Asp Ile 
    560                 565                 570 

aac aaa att gtc caa att cta cca tgg gaa cag aat gag caa gtg aag     1898 
Asn Lys Ile Val Gln Ile Leu Pro Trp Glu Gln Asn Glu Gln Val Lys 
575                 580                 585                 590 

aac ttt gtg gct tcc cat att gcc aat atc ttg aac tca gaa gaa ttg     1946 
Asn Phe Val Ala Ser His Ile Ala Asn Ile Leu Asn Ser Glu Glu Leu 
                595                 600                 605 

gat atc caa gat ctg aaa aag tta gtg aaa gaa gct ctg aaa gaa tct     1994 
Asp Ile Gln Asp Leu Lys Lys Leu Val Lys Glu Ala Leu Lys Glu Ser 
            610                 615                 620 

caa ctt cca act gtc atg gac ttc aga aaa ttc tct cgg aac tat caa     2042 
Gln Leu Pro Thr Val Met Asp Phe Arg Lys Phe Ser Arg Asn Tyr Gln 
        625                 630                 635 

ctc tac aaa tct gtt tct ctt cca tca ctt gac cca gcc tca gcc aaa     2090 
Leu Tyr Lys Ser Val Ser Leu Pro Ser Leu Asp Pro Ala Ser Ala Lys 
    640                 645                 650 

ata gaa ggg aat ctt ata ttt gat cca aat aac tac ctt cct aaa gaa     2138 
Ile Glu Gly Asn Leu Ile Phe Asp Pro Asn Asn Tyr Leu Pro Lys Glu 
655                 660                 665                 670 

agc atg ctg aaa act acc ctc act gcc ttt gga ttt gct tca gct gac     2186 
Ser Met Leu Lys Thr Thr Leu Thr Ala Phe Gly Phe Ala Ser Ala Asp 
                675                 680                 685 

ctc atc gag att ggc ttg gaa gga aaa ggc ttt gag cca aca ttg gaa     2234 
Leu Ile Glu Ile Gly Leu Glu Gly Lys Gly Phe Glu Pro Thr Leu Glu 
            690                 695                 700 

gct ctt ttt ggg aag caa gga ttt ttc cca gac agt gtc aac aaa gct     2282 
Ala Leu Phe Gly Lys Gln Gly Phe Phe Pro Asp Ser Val Asn Lys Ala 
        705                 710                 715 

ttg tac tgg gtt aat ggt caa gtt cct gat ggt gtc tct aag gtc tta     2330 
Leu Tyr Trp Val Asn Gly Gln Val Pro Asp Gly Val Ser Lys Val Leu 
    720                 725                 730 

gtg gac cac ttt ggc tat acc aaa gat gat aaa cat gag cag gat atg     2378 
Val Asp His Phe Gly Tyr Thr Lys Asp Asp Lys His Glu Gln Asp Met 
735                 740                 745                 750 

gta aat gga ata atg ctc agt gtt gag aag ctg att aaa gat ttg aaa     2426 
Val Asn Gly Ile Met Leu Ser Val Glu Lys Leu Ile Lys Asp Leu Lys 
                755                 760                 765 

tcc aaa gaa gtc ccg gaa gcc aga gcc tac ctc cgc atc ttg gga gag     2474 
Ser Lys Glu Val Pro Glu Ala Arg Ala Tyr Leu Arg Ile Leu Gly Glu 
            770                 775                 780 

gag ctt ggt ttt gcc agt ctc cat gac ctc cag ctc ctg gga aag ctg     2522 
Glu Leu Gly Phe Ala Ser Leu His Asp Leu Gln Leu Leu Gly Lys Leu 
        785                 790                 795 

ctt ctg atg ggt gcc cgc act ctg cag ggg atc ccc cag atg att gga     2570 
Leu Leu Met Gly Ala Arg Thr Leu Gln Gly Ile Pro Gln Met Ile Gly 
    800                 805                 810 

gag gtc atc agg aag ggc tca aag aat gac ttt ttt ctt cac tac atc     2618 
Glu Val Ile Arg Lys Gly Ser Lys Asn Asp Phe Phe Leu His Tyr Ile 
815                 820                 825                 830 

ttc atg gag aat gcc ttt gaa ctc ccc act gga gct gga tta cag ttg     2666 
Phe Met Glu Asn Ala Phe Glu Leu Pro Thr Gly Ala Gly Leu Gln Leu 
                835                 840                 845 

caa ata tct tca tct gga gtc att gct ccc gga gcc aag gct gga gta     2714 
Gln Ile Ser Ser Ser Gly Val Ile Ala Pro Gly Ala Lys Ala Gly Val 
            850                 855                 860 

aaa ctg gaa gta gcc aac atg cag gct gaa ctg gtg gca aaa ccc tcc     2762 
Lys Leu Glu Val Ala Asn Met Gln Ala Glu Leu Val Ala Lys Pro Ser 
        865                 870                 875 

gtg tct gtg gag ttt gtg aca aat atg ggc atc atc att ccg gac ttc     2810 
Val Ser Val Glu Phe Val Thr Asn Met Gly Ile Ile Ile Pro Asp Phe 
    880                 885                 890 

gct agg agt ggg gtc cag atg aac acc aac ttc ttc cac gag tcg ggt     2858 
Ala Arg Ser Gly Val Gln Met Asn Thr Asn Phe Phe His Glu Ser Gly 
895                 900                 905                 910 

ctg gag gct cat gtt gcc cta aaa gct ggg aag ctg aag ttt atc att     2906 
Leu Glu Ala His Val Ala Leu Lys Ala Gly Lys Leu Lys Phe Ile Ile 
                915                 920                 925 

cct tcc cca aag aga cca gtc aag ctg ctc agt gga ggc aac aca tta     2954 
Pro Ser Pro Lys Arg Pro Val Lys Leu Leu Ser Gly Gly Asn Thr Leu 
            930                 935                 940 

cat ttg gtc tct acc acc aaa acg gag gtg atc cca cct ctc att gag     3002 
His Leu Val Ser Thr Thr Lys Thr Glu Val Ile Pro Pro Leu Ile Glu 
        945                 950                 955 

aac agg cag tcc tgg tca gtt tgc aag caa gtc ttt cct ggc ctg aat     3050 
Asn Arg Gln Ser Trp Ser Val Cys Lys Gln Val Phe Pro Gly Leu Asn 
    960                 965                 970 

tac tgc acc tca ggc gct tac tcc aac gcc agc tcc aca gac tcc gcc     3098 
Tyr Cys Thr Ser Gly Ala Tyr Ser Asn Ala Ser Ser Thr Asp Ser Ala 
975                 980                 985                 990 

tcc tac tat ccg ctg acc ggg gac acc aga tta gag ctg gaa ctg agg     3146 
Ser Tyr Tyr Pro Leu Thr Gly Asp Thr Arg Leu Glu Leu Glu Leu Arg 
                 995                1000                1005 

cct aca gga gag att gag cag tat tct gtc agc gca acc tat gag ctc     3194 
Pro Thr Gly Glu Ile Glu Gln Tyr Ser Val Ser Ala Thr Tyr Glu Leu 
            1010                1015                1020 

cag aga gag gac aga gcc ttg gtg gat acc ctg aag ttt gta act caa     3242 
Gln Arg Glu Asp Arg Ala Leu Val Asp Thr Leu Lys Phe Val Thr Gln 
        1025                1030                1035 

gca gaa ggt gcg aag cag act gag gct acc atg aca ttc aaa tat aat     3290 
Ala Glu Gly Ala Lys Gln Thr Glu Ala Thr Met Thr Phe Lys Tyr Asn 
    1040                1045                1050 

cgg cag agt atg acc ttg tcc agt gaa gtc caa att ccg gat ttt gat     3338 
Arg Gln Ser Met Thr Leu Ser Ser Glu Val Gln Ile Pro Asp Phe Asp 
1055                1060                1065                1070 

gtt gac ctc gga aca atc ctc aga gtt aat gat gaa tct act gag ggc     3386 
Val Asp Leu Gly Thr Ile Leu Arg Val Asn Asp Glu Ser Thr Glu Gly 
                1075                1080                1085 

aaa acg tct tac aga ctc acc ctg gac att cag aac aag aaa att act     3434 
Lys Thr Ser Tyr Arg Leu Thr Leu Asp Ile Gln Asn Lys Lys Ile Thr 
            1090                1095                1100 

gag gtc gcc ctc atg ggc cac cta agt tgt gac aca aag gaa gaa aga     3482 
Glu Val Ala Leu Met Gly His Leu Ser Cys Asp Thr Lys Glu Glu Arg 
        1105                1110                1115 

aaa atc aag ggt gtt att tcc ata ccc cgt ttg caa gca gaa gcc aga     3530 
Lys Ile Lys Gly Val Ile Ser Ile Pro Arg Leu Gln Ala Glu Ala Arg 
    1120                1125                1130 

agt gag atc ctc gcc cac tgg tcg cct gcc aaa ctg ctt ctc caa atg     3578 
Ser Glu Ile Leu Ala His Trp Ser Pro Ala Lys Leu Leu Leu Gln Met 
1135                1140                1145                1150 

gac tca tct gct aca gct tat ggc tcc aca gtt tcc aag agg gtg gca     3626 
Asp Ser Ser Ala Thr Ala Tyr Gly Ser Thr Val Ser Lys Arg Val Ala 
                1155                1160                1165 

tgg cat tat gat gaa gag aag att gaa ttt gaa tgg aac aca ggc acc     3674 
Trp His Tyr Asp Glu Glu Lys Ile Glu Phe Glu Trp Asn Thr Gly Thr 
            1170                1175                1180 

aat gta gat acc aaa aaa atg act tcc aat ttc cct gtg gat ctc tcc     3722 
Asn Val Asp Thr Lys Lys Met Thr Ser Asn Phe Pro Val Asp Leu Ser 
        1185                1190                1195 

gat tat cct aag agc ttg cat atg tat gct aat aga ctc ctg gat cac     3770 
Asp Tyr Pro Lys Ser Leu His Met Tyr Ala Asn Arg Leu Leu Asp His 
    1200                1205                1210 

aga gtc cct gaa aca gac atg act ttc cgg cac gtg ggt tcc aaa tta     3818 
Arg Val Pro Glu Thr Asp Met Thr Phe Arg His Val Gly Ser Lys Leu 
1215                1220                1225                1230 

ata gtt gca atg agc tca tgg ctt cag aag gca tct ggg agt ctt cct     3866 
Ile Val Ala Met Ser Ser Trp Leu Gln Lys Ala Ser Gly Ser Leu Pro 
                1235                1240                1245 

tat acc cag act ttg caa gac cac ctc aat agc ctg aag gag ttc aac     3914 
Tyr Thr Gln Thr Leu Gln Asp His Leu Asn Ser Leu Lys Glu Phe Asn 
            1250                1255                1260 

ctc cag aac atg gga ttg cca gac ttc cac atc cca gaa aac ctc ttc     3962 
Leu Gln Asn Met Gly Leu Pro Asp Phe His Ile Pro Glu Asn Leu Phe 
        1265                1270                1275 

tta aaa agc gat ggc cgg gtc aaa tat acc ttg aac aag aac agt ttg     4010 
Leu Lys Ser Asp Gly Arg Val Lys Tyr Thr Leu Asn Lys Asn Ser Leu 
    1280                1285                1290 

aaa att gag att cct ttg cct ttt ggt ggc aaa tcc tcc aga gat cta     4058 
Lys Ile Glu Ile Pro Leu Pro Phe Gly Gly Lys Ser Ser Arg Asp Leu 
1295                1300                1305                1310 

aag atg tta gag act gtt agg aca cca gcc ctc cac ttc aag tct gtg     4106 
Lys Met Leu Glu Thr Val Arg Thr Pro Ala Leu His Phe Lys Ser Val 
                1315                1320                1325 

gga ttc cat ctg cca tct cga gag ttc caa gtc cct act ttt acc att     4154 
Gly Phe His Leu Pro Ser Arg Glu Phe Gln Val Pro Thr Phe Thr Ile 
            1330                1335                1340 

ccc aag ttg tat caa ctg caa gtg cct ctc ctg ggt gtt cta gac ctc     4202 
Pro Lys Leu Tyr Gln Leu Gln Val Pro Leu Leu Gly Val Leu Asp Leu 
        1345                1350                1355 

tcc acg aat gtc tac agc aac ttg tac aac tgg tcc gcc tcc tac agt     4250 
Ser Thr Asn Val Tyr Ser Asn Leu Tyr Asn Trp Ser Ala Ser Tyr Ser 
    1360                1365                1370 

ggt ggc aac acc agc aca gac cat ttc agc ctt cgg gct cgt tac cac     4298 
Gly Gly Asn Thr Ser Thr Asp His Phe Ser Leu Arg Ala Arg Tyr His 
1375                1380                1385                1390 

atg aag gct gac tct gtg gtt gac ctg ctt tcc tac aat gtg caa gga     4346 
Met Lys Ala Asp Ser Val Val Asp Leu Leu Ser Tyr Asn Val Gln Gly 
                1395                1400                1405 

tct gga gaa aca aca tat gac cac aag aat acg ttc aca cta tca tgt     4394 
Ser Gly Glu Thr Thr Tyr Asp His Lys Asn Thr Phe Thr Leu Ser Cys 
            1410                1415                1420 

gat ggg tct cta cgc cac aaa ttt cta gat tcg aat atc aaa ttc agt     4442 
Asp Gly Ser Leu Arg His Lys Phe Leu Asp Ser Asn Ile Lys Phe Ser 
        1425                1430                1435 

cat gta gaa aaa ctt gga aac aac cca gtc tca aaa ggt tta cta ata     4490 
His Val Glu Lys Leu Gly Asn Asn Pro Val Ser Lys Gly Leu Leu Ile 
    1440                1445                1450 

ttc gat gca tct agt tcc tgg gga cca cag atg tct gct tca gtt cat     4538 
Phe Asp Ala Ser Ser Ser Trp Gly Pro Gln Met Ser Ala Ser Val His 
1455                1460                1465                1470 

ttg gac tcc aaa aag aaa cag cat ttg ttt gtc aaa gaa gtc aag att     4586 
Leu Asp Ser Lys Lys Lys Gln His Leu Phe Val Lys Glu Val Lys Ile 
                1475                1480                1485 

gat ggg cag ttc aga gtc tct tcg ttc tat gct aaa ggc aca tat ggc     4634 
Asp Gly Gln Phe Arg Val Ser Ser Phe Tyr Ala Lys Gly Thr Tyr Gly 
            1490                1495                1500 

ctg tct tgt cag agg gat cct aac act ggc cgg ctc aat gga gag tcc     4682 
Leu Ser Cys Gln Arg Asp Pro Asn Thr Gly Arg Leu Asn Gly Glu Ser 
        1505                1510                1515 

aac ctg agg ttt aac tcc tcc tac ctc caa ggc acc aac cag ata aca     4730 
Asn Leu Arg Phe Asn Ser Ser Tyr Leu Gln Gly Thr Asn Gln Ile Thr 
    1520                1525                1530 

gga aga tat gaa gat gga acc ctc tcc ctc acc tcc acc tct gat ctg     4778 
Gly Arg Tyr Glu Asp Gly Thr Leu Ser Leu Thr Ser Thr Ser Asp Leu 
1535                1540                1545                1550 

caa agt ggc atc att aaa aat act gct tcc cta aag tat gag aac tac     4826 
Gln Ser Gly Ile Ile Lys Asn Thr Ala Ser Leu Lys Tyr Glu Asn Tyr 
                1555                1560                1565 

gag ctg act tta aaa tct gac acc aat ggg aag tat aag aac ttt gcc     4874 
Glu Leu Thr Leu Lys Ser Asp Thr Asn Gly Lys Tyr Lys Asn Phe Ala 
            1570                1575                1580 

act tct aac aag atg gat atg acc ttc tct aag caa aat gca ctg ctg     4922 
Thr Ser Asn Lys Met Asp Met Thr Phe Ser Lys Gln Asn Ala Leu Leu 
        1585                1590                1595 

cgt tct gaa tat cag gct gat tac gag tca ttg agg ttc ttc agc ctg     4970 
Arg Ser Glu Tyr Gln Ala Asp Tyr Glu Ser Leu Arg Phe Phe Ser Leu 
    1600                1605                1610 

ctt tct gga tca cta aat tcc cat ggt ctt gag tta aat gct gac atc     5018 
Leu Ser Gly Ser Leu Asn Ser His Gly Leu Glu Leu Asn Ala Asp Ile 
1615                1620                1625                1630 

tta ggc act gac aaa att aat agt ggt gct cac aag gcg aca cta agg     5066 
Leu Gly Thr Asp Lys Ile Asn Ser Gly Ala His Lys Ala Thr Leu Arg 
                1635                1640                1645 

att ggc caa gat gga ata tct acc agt gca acg acc aac ttg aag tgt     5114 
Ile Gly Gln Asp Gly Ile Ser Thr Ser Ala Thr Thr Asn Leu Lys Cys 
            1650                1655                1660 

agt ctc ctg gtg ctg gag aat gag ctg aat gca gag ctt ggc ctc tct     5162 
Ser Leu Leu Val Leu Glu Asn Glu Leu Asn Ala Glu Leu Gly Leu Ser 
        1665                1670                1675 

ggg gca tct atg aaa tta aca aca aat ggc cgc ttc agg gaa cac aat     5210 
Gly Ala Ser Met Lys Leu Thr Thr Asn Gly Arg Phe Arg Glu His Asn 
    1680                1685                1690 

gca aaa ttc agt ctg gat ggg aaa gcc gcc ctc aca gag cta tca ctg     5258 
Ala Lys Phe Ser Leu Asp Gly Lys Ala Ala Leu Thr Glu Leu Ser Leu 
1695                1700                1705                1710 

gga agt gct tat cag gcc atg att ctg ggt gtc gac agc aaa aac att     5306 
Gly Ser Ala Tyr Gln Ala Met Ile Leu Gly Val Asp Ser Lys Asn Ile 
                1715                1720                1725 

ttc aac ttc aag gtc agt caa gaa gga ctt aag ctc tca aat gac atg     5354 
Phe Asn Phe Lys Val Ser Gln Glu Gly Leu Lys Leu Ser Asn Asp Met 
            1730                1735                1740 

atg ggc tca tat gct gaa atg aaa ttt gac cac aca aac agt ctg aac     5402 
Met Gly Ser Tyr Ala Glu Met Lys Phe Asp His Thr Asn Ser Leu Asn 
        1745                1750                1755 

att gca ggc tta tca ctg gac ttc tct tca aaa ctt gac aac att tac     5450 
Ile Ala Gly Leu Ser Leu Asp Phe Ser Ser Lys Leu Asp Asn Ile Tyr 
    1760                1765                1770 

agc tct gac aag ttt tat aag caa act gtt aat tta cag cta cag ccc     5498 
Ser Ser Asp Lys Phe Tyr Lys Gln Thr Val Asn Leu Gln Leu Gln Pro 
1775                1780                1785                1790 

tat tct ctg gta act act tta aac agt gac ctg aaa tac aat gct ctg     5546 
Tyr Ser Leu Val Thr Thr Leu Asn Ser Asp Leu Lys Tyr Asn Ala Leu 
                1795                1800                1805 

gat ctc acc aac aat ggg aaa cta cgg cta gaa ccc ctg aag ctg cat     5594 
Asp Leu Thr Asn Asn Gly Lys Leu Arg Leu Glu Pro Leu Lys Leu His 
            1810                1815                1820 

gtg gct ggt aac cta aaa gga gcc tac caa aat aat gaa ata aaa cac     5642 
Val Ala Gly Asn Leu Lys Gly Ala Tyr Gln Asn Asn Glu Ile Lys His 
        1825                1830                1835 

atc tat gcc atc tct tct gct gcc tta tca gca agc tat aaa gca gac     5690 
Ile Tyr Ala Ile Ser Ser Ala Ala Leu Ser Ala Ser Tyr Lys Ala Asp 
    1840                1845                1850 

act gtt gct aag gtt cag ggt gtg gag ttt agc cat cgg ctc aac aca     5738 
Thr Val Ala Lys Val Gln Gly Val Glu Phe Ser His Arg Leu Asn Thr 
1855                1860                1865                1870 

gac atc gct ggg ctg gct tca gcc att gac atg agc aca aac tat aat     5786 
Asp Ile Ala Gly Leu Ala Ser Ala Ile Asp Met Ser Thr Asn Tyr Asn 
                1875                1880                1885 

tca gac tca ctg cat ttc agc aat gtc ttc cgt tct gta atg gcc ccg     5834 
Ser Asp Ser Leu His Phe Ser Asn Val Phe Arg Ser Val Met Ala Pro 
            1890                1895                1900 

ttt acc atg acc atc gat gca cat aca aat ggc aat ggg aaa ctc gct     5882 
Phe Thr Met Thr Ile Asp Ala His Thr Asn Gly Asn Gly Lys Leu Ala 
        1905                1910                1915 

ctc tgg gga gaa cat act ggg cag ctg tat agc aaa ttc ctg ttg aaa     5930 
Leu Trp Gly Glu His Thr Gly Gln Leu Tyr Ser Lys Phe Leu Leu Lys 
    1920                1925                1930 

gca gaa cct ctg gca ttt act ttc tct cat gat tac aaa ggc tcc aca     5978 
Ala Glu Pro Leu Ala Phe Thr Phe Ser His Asp Tyr Lys Gly Ser Thr 
1935                1940                1945                1950 

agt cat cat ctc gtg tct agg aaa agc atc agt gca gct ctt gaa cac     6026 
Ser His His Leu Val Ser Arg Lys Ser Ile Ser Ala Ala Leu Glu His 
                1955                1960                1965 

aaa gtc agt gcc ctg ctt act cca gct gag cag aca ggc acc tgg aaa     6074 
Lys Val Ser Ala Leu Leu Thr Pro Ala Glu Gln Thr Gly Thr Trp Lys 
            1970                1975                1980 

ctc aag acc caa ttt aac aac aat gaa tac agc cag gac ttg gat gct     6122 
Leu Lys Thr Gln Phe Asn Asn Asn Glu Tyr Ser Gln Asp Leu Asp Ala 
        1985                1990                1995 

tac aac act aaa gat aaa att ggc gtg gag ctt act gga cga act ctg     6170 
Tyr Asn Thr Lys Asp Lys Ile Gly Val Glu Leu Thr Gly Arg Thr Leu 
    2000                2005                2010 

gct gac cta act cta cta gac tcc cca att aaa gtg cca ctt tta ctc     6218 
Ala Asp Leu Thr Leu Leu Asp Ser Pro Ile Lys Val Pro Leu Leu Leu 
2015                2020                2025                2030 

agt gag ccc atc aat atc att gat gct tta gag atg aga gat gcc gtt     6266 
Ser Glu Pro Ile Asn Ile Ile Asp Ala Leu Glu Met Arg Asp Ala Val 
                2035                2040                2045 

gag aag ccc caa gaa ttt aca att gtt gct ttt gta aag tat gat aaa     6314 
Glu Lys Pro Gln Glu Phe Thr Ile Val Ala Phe Val Lys Tyr Asp Lys 
            2050                2055                2060 

aac caa gat gtt cac tcc att aac ctc cca ttt ttt gag acc ttg caa     6362 
Asn Gln Asp Val His Ser Ile Asn Leu Pro Phe Phe Glu Thr Leu Gln 
        2065                2070                2075 

gaa tat ttt gag agg aat cga caa acc att ata gtt gta gtg gaa aac     6410 
Glu Tyr Phe Glu Arg Asn Arg Gln Thr Ile Ile Val Val Val Glu Asn 
    2080                2085                2090 

gta cag aga aac ctg aag cac atc aat att gat caa ttt gta aga aaa     6458 
Val Gln Arg Asn Leu Lys His Ile Asn Ile Asp Gln Phe Val Arg Lys 
2095                2100                2105                2110 

tac aga gca gcc ctg gga aaa ctc cca cag caa gct aat gat tat ctg     6506 
Tyr Arg Ala Ala Leu Gly Lys Leu Pro Gln Gln Ala Asn Asp Tyr Leu 
                2115                2120                2125 

aat tca ttc aat tgg gag aga caa gtt tca cat gcc aag gag aaa ctg     6554 
Asn Ser Phe Asn Trp Glu Arg Gln Val Ser His Ala Lys Glu Lys Leu 
            2130                2135                2140 

act gct ctc aca aaa aag tat aga att aca gaa aat gat ata caa att     6602 
Thr Ala Leu Thr Lys Lys Tyr Arg Ile Thr Glu Asn Asp Ile Gln Ile 
        2145                2150                2155 

gca tta gat gat gcc aaa atc aac ttt aat gaa aaa cta tct caa ctg     6650 
Ala Leu Asp Asp Ala Lys Ile Asn Phe Asn Glu Lys Leu Ser Gln Leu 
    2160                2165                2170 

cag aca tat atg ata caa ttt gat cag tat att aaa gat agt tat gat     6698 
Gln Thr Tyr Met Ile Gln Phe Asp Gln Tyr Ile Lys Asp Ser Tyr Asp 
2175                2180                2185                2190 

tta cat gat ttg aaa ata gct att gct aat att att gat gaa atc att     6746 
Leu His Asp Leu Lys Ile Ala Ile Ala Asn Ile Ile Asp Glu Ile Ile 
                2195                2200                2205 

gaa aaa tta aaa agt ctt gat gag cac tat cat atc cgt gta aat tta     6794 
Glu Lys Leu Lys Ser Leu Asp Glu His Tyr His Ile Arg Val Asn Leu 
            2210                2215                2220 

gta aaa aca atc cat gat cta cat ttg ttt att gaa aat att gat ttt     6842 
Val Lys Thr Ile His Asp Leu His Leu Phe Ile Glu Asn Ile Asp Phe 
        2225                2230                2235 

aac aaa agt gga agt agt act gca tcc tgg att caa aat gtg gat act     6890 
Asn Lys Ser Gly Ser Ser Thr Ala Ser Trp Ile Gln Asn Val Asp Thr 
    2240                2245                2250 

aag tac caa atc aga atc cag ata caa gaa aaa ctg cag cag ctt aag     6938 
Lys Tyr Gln Ile Arg Ile Gln Ile Gln Glu Lys Leu Gln Gln Leu Lys 
2255                2260                2265                2270 

aga cac ata cag aat ata gac atc cag cac cta gct gga aag tta aaa     6986 
Arg His Ile Gln Asn Ile Asp Ile Gln His Leu Ala Gly Lys Leu Lys 
                2275                2280                2285 

caa cac att gag gct att gat gtt aga gtg ctt tta gat caa ttg gga     7034 
Gln His Ile Glu Ala Ile Asp Val Arg Val Leu Leu Asp Gln Leu Gly 
            2290                2295                2300 

act aca att tca ttt gaa aga ata aat gat gtt ctt gag cat gtc aaa     7082 
Thr Thr Ile Ser Phe Glu Arg Ile Asn Asp Val Leu Glu His Val Lys 
        2305                2310                2315 

cac ttt gtt ata aat ctt att ggg gat ttt gaa gta gct gag aaa atc     7130 
His Phe Val Ile Asn Leu Ile Gly Asp Phe Glu Val Ala Glu Lys Ile 
    2320                2325                2330 

aat gcc ttc aga gcc aaa gtc cat gag tta atc gag agg tat gaa gta     7178 
Asn Ala Phe Arg Ala Lys Val His Glu Leu Ile Glu Arg Tyr Glu Val 
2335                2340                2345                2350 

gac caa caa atc cag gtt tta atg gat aaa tta gta gag ttg acc cac     7226 
Asp Gln Gln Ile Gln Val Leu Met Asp Lys Leu Val Glu Leu Thr His 
                2355                2360                2365 

caa tac aag ttg aag gag act att cag aag cta agc aat gtc cta caa     7274 
Gln Tyr Lys Leu Lys Glu Thr Ile Gln Lys Leu Ser Asn Val Leu Gln 
            2370                2375                2380 

caa gtt aag ata aaa gat tac ttt gag aaa ttg gtt gga ttt att gat     7322 
Gln Val Lys Ile Lys Asp Tyr Phe Glu Lys Leu Val Gly Phe Ile Asp 
        2385                2390                2395 

gat gct gtg aag aag ctt aat gaa tta tct ttt aaa aca ttc att gaa     7370 
Asp Ala Val Lys Lys Leu Asn Glu Leu Ser Phe Lys Thr Phe Ile Glu 
    2400                2405                2410 

gat gtt aac aaa ttc ctt gac atg ttg ata aag aaa tta aag tca ttt     7418 
Asp Val Asn Lys Phe Leu Asp Met Leu Ile Lys Lys Leu Lys Ser Phe 
2415                2420                2425                2430 

gat tac cac cag ttt gta gat gaa acc aat gac aaa atc cgt gag gtg     7466 
Asp Tyr His Gln Phe Val Asp Glu Thr Asn Asp Lys Ile Arg Glu Val 
                2435                2440                2445 

act cag aga ctc aat ggt gaa att cag gct ctg gaa cta cca caa aaa     7514 
Thr Gln Arg Leu Asn Gly Glu Ile Gln Ala Leu Glu Leu Pro Gln Lys 
            2450                2455                2460 

gct gaa gca tta aaa ctg ttt tta gag gaa acc aag gcc aca gtt gca     7562 
Ala Glu Ala Leu Lys Leu Phe Leu Glu Glu Thr Lys Ala Thr Val Ala 
        2465                2470                2475 

gtg tat ctg gaa agc cta cag gac acc aaa ata acc tta atc atc aat     7610 
Val Tyr Leu Glu Ser Leu Gln Asp Thr Lys Ile Thr Leu Ile Ile Asn 
    2480                2485                2490 

tgg tta cag gag gct tta agt tca gca tct ttg gct cac atg aag gcc     7658 
Trp Leu Gln Glu Ala Leu Ser Ser Ala Ser Leu Ala His Met Lys Ala 
2495                2500                2505                2510 

aaa ttc cga gag act cta gaa gat aca cga gac cga atg tat caa atg     7706 
Lys Phe Arg Glu Thr Leu Glu Asp Thr Arg Asp Arg Met Tyr Gln Met 
                2515                2520                2525 

gac att cag cag gaa ctt caa cga tac ctg tct ctg gta ggc cag gtt     7754 
Asp Ile Gln Gln Glu Leu Gln Arg Tyr Leu Ser Leu Val Gly Gln Val 
            2530                2535                2540 

tat agc aca ctt gtc acc tac att tct gat tgg tgg act ctt gct gct     7802 
Tyr Ser Thr Leu Val Thr Tyr Ile Ser Asp Trp Trp Thr Leu Ala Ala 
        2545                2550                2555 

aag aac ctt act gac ttt gca gag caa tat tct atc caa gat tgg gct     7850 
Lys Asn Leu Thr Asp Phe Ala Glu Gln Tyr Ser Ile Gln Asp Trp Ala 
    2560                2565                2570 

aaa cgt atg aaa gca ttg gta gag caa ggg ttc act gtt cct gaa atc     7898 
Lys Arg Met Lys Ala Leu Val Glu Gln Gly Phe Thr Val Pro Glu Ile 
2575                2580                2585                2590 

aag acc atc ctt ggg acc atg cct gcc ttt gaa gtc agt ctt cag gct     7946 
Lys Thr Ile Leu Gly Thr Met Pro Ala Phe Glu Val Ser Leu Gln Ala 
                2595                2600                2605 

ctt cag aaa gct acc ttc cag aca cct gat ttt ata gtc ccc cta aca     7994 
Leu Gln Lys Ala Thr Phe Gln Thr Pro Asp Phe Ile Val Pro Leu Thr 
            2610                2615                2620 

gat ttg agg att cca tca gtt cag ata aac ttc aaa gac tta aaa aat     8042 
Asp Leu Arg Ile Pro Ser Val Gln Ile Asn Phe Lys Asp Leu Lys Asn 
        2625                2630                2635 

ata aaa atc cca tcc agg ttt tcc aca cca gaa ttt acc atc ctt aac     8090 
Ile Lys Ile Pro Ser Arg Phe Ser Thr Pro Glu Phe Thr Ile Leu Asn 
    2640                2645                2650 

acc ttc cac att cct tcc ttt aca att gac ttt gtc gaa atg aaa gta     8138 
Thr Phe His Ile Pro Ser Phe Thr Ile Asp Phe Val Glu Met Lys Val 
2655                2660                2665                2670 

aag atc atc aga acc att gac cag atg cag aac agt gag ctg cag tgg     8186 
Lys Ile Ile Arg Thr Ile Asp Gln Met Gln Asn Ser Glu Leu Gln Trp 
                2675                2680                2685 

ccc gtt cca gat ata tat ctc agg gat ctg aag gtg gag gac att cct     8234 
Pro Val Pro Asp Ile Tyr Leu Arg Asp Leu Lys Val Glu Asp Ile Pro 
            2690                2695                2700 

cta gcg aga atc acc ctg cca gac ttc cgt tta cca gaa atc gca att     8282 
Leu Ala Arg Ile Thr Leu Pro Asp Phe Arg Leu Pro Glu Ile Ala Ile 
        2705                2710                2715 

cca gaa ttc ata atc cca act ctc aac ctt aat gat ttt caa gtt cct     8330 
Pro Glu Phe Ile Ile Pro Thr Leu Asn Leu Asn Asp Phe Gln Val Pro 
    2720                2725                2730 

gac ctt cac ata cca gaa ttc cag ctt ccc cac atc tca cac aca att     8378 
Asp Leu His Ile Pro Glu Phe Gln Leu Pro His Ile Ser His Thr Ile 
2735                2740                2745                2750 

gaa gta cct act ttt ggc aag cta tac agt att ctg aaa atc caa tct     8426 
Glu Val Pro Thr Phe Gly Lys Leu Tyr Ser Ile Leu Lys Ile Gln Ser 
                2755                2760                2765 

cct ctt ttc aca tta gat gca aat gct gac ata ggg aat gga acc acc     8474 
Pro Leu Phe Thr Leu Asp Ala Asn Ala Asp Ile Gly Asn Gly Thr Thr 
            2770                2775                2780 

tca gca aac gaa gca ggt atc gca gct tcc atc act gcc aaa gga gag     8522 
Ser Ala Asn Glu Ala Gly Ile Ala Ala Ser Ile Thr Ala Lys Gly Glu 
        2785                2790                2795 

tcc aaa tta gaa gtt ctc aat ttt gat ttt caa gca aat gca caa ctc     8570 
Ser Lys Leu Glu Val Leu Asn Phe Asp Phe Gln Ala Asn Ala Gln Leu 
    2800                2805                2810 

tca aac cct aag att aat ccg ctg gct ctg aag gag tca gtg aag ttc     8618 
Ser Asn Pro Lys Ile Asn Pro Leu Ala Leu Lys Glu Ser Val Lys Phe 
2815                2820                2825                2830 

tcc agc aag tac ctg aga acg gag cat ggg agt gaa atg ctg ttt ttt     8666 
Ser Ser Lys Tyr Leu Arg Thr Glu His Gly Ser Glu Met Leu Phe Phe 
                2835                2840                2845 

gga aat gct att gag gga aaa tca aac aca gtg gca agt tta cac aca     8714 
Gly Asn Ala Ile Glu Gly Lys Ser Asn Thr Val Ala Ser Leu His Thr 
            2850                2855                2860 

gaa aaa aat aca ctg gag ctt agt aat gga gtg att gtc aag ata aac     8762 
Glu Lys Asn Thr Leu Glu Leu Ser Asn Gly Val Ile Val Lys Ile Asn 
        2865                2870                2875 

aat cag ctt acc ctg gat agc aac act aaa tac ttc cac aaa ttg aac     8810 
Asn Gln Leu Thr Leu Asp Ser Asn Thr Lys Tyr Phe His Lys Leu Asn 
    2880                2885                2890 

atc ccc aaa ctg gac ttc tct agt cag gct gac ctg cgc aac gag atc     8858 
Ile Pro Lys Leu Asp Phe Ser Ser Gln Ala Asp Leu Arg Asn Glu Ile 
2895                2900                2905                2910 

aag aca ctg ttg aaa gct ggc cac ata gca tgg act tct tct gga aaa     8906 
Lys Thr Leu Leu Lys Ala Gly His Ile Ala Trp Thr Ser Ser Gly Lys 
                2915                2920                2925 

ggg tca tgg aaa tgg gcc tgc ccc aga ttc tca gat gag gga aca cat     8954 
Gly Ser Trp Lys Trp Ala Cys Pro Arg Phe Ser Asp Glu Gly Thr His 
            2930                2935                2940 

gaa tca caa att agt ttc acc ata gaa gga ccc ctc act tcc ttt gga     9002 
Glu Ser Gln Ile Ser Phe Thr Ile Glu Gly Pro Leu Thr Ser Phe Gly 
        2945                2950                2955 

ctg tcc aat aag atc aat agc aaa cac cta aga gta aac caa aac ttg     9050 
Leu Ser Asn Lys Ile Asn Ser Lys His Leu Arg Val Asn Gln Asn Leu 
    2960                2965                2970 

gtt tat gaa tct ggc tcc ctc aac ttt tct aaa ctt gaa att caa tca     9098 
Val Tyr Glu Ser Gly Ser Leu Asn Phe Ser Lys Leu Glu Ile Gln Ser 
2975                2980                2985                2990 

caa gtc gat tcc cag cat gtg ggc cac agt gtt cta act gct aaa ggc     9146 
Gln Val Asp Ser Gln His Val Gly His Ser Val Leu Thr Ala Lys Gly 
                2995                3000                3005 

atg gca ctg ttt gga gaa ggg aag gca gag ttt act ggg agg cat gat     9194 
Met Ala Leu Phe Gly Glu Gly Lys Ala Glu Phe Thr Gly Arg His Asp 
            3010                3015                3020 

gct cat tta aat gga aag gtt att gga act ttg aaa aat tct ctt ttc     9242 
Ala His Leu Asn Gly Lys Val Ile Gly Thr Leu Lys Asn Ser Leu Phe 
        3025                3030                3035 

ttt tca gcc cag cca ttt gag atc acg gca tcc aca aac aat gaa ggg     9290 
Phe Ser Ala Gln Pro Phe Glu Ile Thr Ala Ser Thr Asn Asn Glu Gly 
    3040                3045                3050 

aat ttg aaa gtt cgt ttt cca tta agg tta aca ggg aag ata gac ttc     9338 
Asn Leu Lys Val Arg Phe Pro Leu Arg Leu Thr Gly Lys Ile Asp Phe 
3055                3060                3065                3070 

ctg aat aac tat gca ctg ttt ctg agt ccc agt gcc cag caa gca agt     9386 
Leu Asn Asn Tyr Ala Leu Phe Leu Ser Pro Ser Ala Gln Gln Ala Ser 
                3075                3080                3085 

tgg caa gta agt gct agg ttc aat cag tat aag tac aac caa aat ttc     9434 
Trp Gln Val Ser Ala Arg Phe Asn Gln Tyr Lys Tyr Asn Gln Asn Phe 
            3090                3095                3100 

tct gct gga aac aac gag aac att atg gag gcc cat gta gga ata aat     9482 
Ser Ala Gly Asn Asn Glu Asn Ile Met Glu Ala His Val Gly Ile Asn 
        3105                3110                3115 

gga gaa gca aat ctg gat ttc tta aac att cct tta aca att cct gaa     9530 
Gly Glu Ala Asn Leu Asp Phe Leu Asn Ile Pro Leu Thr Ile Pro Glu 
    3120                3125                3130 

atg cgt cta cct tac aca ata atc aca act cct cca ctg aaa gat ttc     9578 
Met Arg Leu Pro Tyr Thr Ile Ile Thr Thr Pro Pro Leu Lys Asp Phe 
3135                3140                3145                3150 

tct cta tgg gaa aaa aca ggc ttg aag gaa ttc ttg aaa acg aca aag     9626 
Ser Leu Trp Glu Lys Thr Gly Leu Lys Glu Phe Leu Lys Thr Thr Lys 
                3155                3160                3165 

caa tca ttt gat tta agt gta aaa gct cag tat aag aaa aac aaa cac     9674 
Gln Ser Phe Asp Leu Ser Val Lys Ala Gln Tyr Lys Lys Asn Lys His 
            3170                3175                3180 

agg cat tcc atc aca aat cct ttg gct gtg ctt tgt gag ttt atc agt     9722 
Arg His Ser Ile Thr Asn Pro Leu Ala Val Leu Cys Glu Phe Ile Ser 
        3185                3190                3195 

cag agc atc aaa tcc ttt gac agg cat ttt gaa aaa aac aga aac aat     9770 
Gln Ser Ile Lys Ser Phe Asp Arg His Phe Glu Lys Asn Arg Asn Asn 
    3200                3205                3210 

gca tta gat ttt gtc acc aaa tcc tat aat gaa aca aaa att aag ttt     9818 
Ala Leu Asp Phe Val Thr Lys Ser Tyr Asn Glu Thr Lys Ile Lys Phe 
3215                3220                3225                3230 

gat aag tac aaa gct gaa aaa tct cac gac gag ctc ccc agg acc ttt     9866 
Asp Lys Tyr Lys Ala Glu Lys Ser His Asp Glu Leu Pro Arg Thr Phe 
                3235                3240                3245 

caa att cct gga tac act gtt cca gtt gtc aat gtt gaa gtg tct cca     9914 
Gln Ile Pro Gly Tyr Thr Val Pro Val Val Asn Val Glu Val Ser Pro 
            3250                3255                3260 

ttc acc ata gag atg tcg gca ttc ggc tat gtg ttc cca aaa gca gtc     9962 
Phe Thr Ile Glu Met Ser Ala Phe Gly Tyr Val Phe Pro Lys Ala Val 
        3265                3270                3275 

agc atg cct agt ttc tcc atc cta ggt tct gac gtc cgt gtg cct tca    10010 
Ser Met Pro Ser Phe Ser Ile Leu Gly Ser Asp Val Arg Val Pro Ser 
    3280                3285                3290 

tac aca tta atc ctg cca tca tta gag ctg cca gtc ctt cat gtc cct    10058 
Tyr Thr Leu Ile Leu Pro Ser Leu Glu Leu Pro Val Leu His Val Pro 
3295                3300                3305                3310 

aga aat ctc aag ctt tct ctt cca cat ttc aag gaa ttg tgt acc ata    10106 
Arg Asn Leu Lys Leu Ser Leu Pro His Phe Lys Glu Leu Cys Thr Ile 
                3315                3320                3325 

agc cat att ttt att cct gcc atg ggc aat att acc tat gat ttc tcc    10154 
Ser His Ile Phe Ile Pro Ala Met Gly Asn Ile Thr Tyr Asp Phe Ser 
            3330                3335                3340 

ttt aaa tca agt gtc atc aca ctg aat acc aat gct gaa ctt ttt aac    10202 
Phe Lys Ser Ser Val Ile Thr Leu Asn Thr Asn Ala Glu Leu Phe Asn 
        3345                3350                3355 

cag tca gat att gtt gct cat ctc ctt tct tca tct tca tct gtc att    10250 
Gln Ser Asp Ile Val Ala His Leu Leu Ser Ser Ser Ser Ser Val Ile 
    3360                3365                3370 

gat gca ctg cag tac aaa tta gag ggc acc aca aga ttg aca aga aaa    10298 
Asp Ala Leu Gln Tyr Lys Leu Glu Gly Thr Thr Arg Leu Thr Arg Lys 
3375                3380                3385                3390 

agg gga ttg aag tta gcc aca gct ctg tct ctg agc aac aaa ttt gtg    10346 
Arg Gly Leu Lys Leu Ala Thr Ala Leu Ser Leu Ser Asn Lys Phe Val 
                3395                3400                3405 

gag ggt agt cat aac agt act gtg agc tta acc acg aaa aat atg gaa    10394 
Glu Gly Ser His Asn Ser Thr Val Ser Leu Thr Thr Lys Asn Met Glu 
            3410                3415                3420 

gtg tca gtg gca aaa acc aca aaa gcc gaa att cca att ttg aga atg    10442 
Val Ser Val Ala Lys Thr Thr Lys Ala Glu Ile Pro Ile Leu Arg Met 
        3425                3430                3435 

aat ttc aag caa gaa ctt aat gga aat acc aag tca aaa cct act gtc    10490 
Asn Phe Lys Gln Glu Leu Asn Gly Asn Thr Lys Ser Lys Pro Thr Val 
    3440                3445                3450 

tct tcc tcc atg gaa ttt aag tat gat ttc aat tct tca atg ctg tac    10538 
Ser Ser Ser Met Glu Phe Lys Tyr Asp Phe Asn Ser Ser Met Leu Tyr 
3455                3460                3465                3470 

tct acc gct aaa gga gca gtt gac cac aag ctt agc ttg gaa agc ctc    10586 
Ser Thr Ala Lys Gly Ala Val Asp His Lys Leu Ser Leu Glu Ser Leu 
                3475                3480                3485 

acc tct tac ttt tcc att gag tca tct acc aaa gga gat gtc aag ggt    10634 
Thr Ser Tyr Phe Ser Ile Glu Ser Ser Thr Lys Gly Asp Val Lys Gly 
            3490                3495                3500 

tcg gtt ctt tct cgg gaa tat tca gga act att gct agt gag gcc aac    10682 
Ser Val Leu Ser Arg Glu Tyr Ser Gly Thr Ile Ala Ser Glu Ala Asn 
        3505                3510                3515 

act tac ttg aat tcc aag agc aca cgg tct tca gtg aag ctg cag ggc    10730 
Thr Tyr Leu Asn Ser Lys Ser Thr Arg Ser Ser Val Lys Leu Gln Gly 
    3520                3525                3530 

act tcc aaa att gat gat atc tgg aac ctt gaa gta aaa gaa aat ttt    10778 
Thr Ser Lys Ile Asp Asp Ile Trp Asn Leu Glu Val Lys Glu Asn Phe 
3535                3540                3545                3550 

gct gga gaa gcc aca ctc caa cgc ata tat tcc ctc tgg gag cac agt    10826 
Ala Gly Glu Ala Thr Leu Gln Arg Ile Tyr Ser Leu Trp Glu His Ser 
                3555                3560                3565 

acg aaa aac cac tta cag cta gag ggc ctc ttt ttc acc aac gga gaa    10874 
Thr Lys Asn His Leu Gln Leu Glu Gly Leu Phe Phe Thr Asn Gly Glu 
            3570                3575                3580 

cat aca agc aaa gcc acc ctg gaa ctc tct cca tgg caa atg tca gct    10922 
His Thr Ser Lys Ala Thr Leu Glu Leu Ser Pro Trp Gln Met Ser Ala 
        3585                3590                3595 

ctt gtt cag gtc cat gca agt cag ccc agt tcc ttc cat gat ttc cct    10970 
Leu Val Gln Val His Ala Ser Gln Pro Ser Ser Phe His Asp Phe Pro 
    3600                3605                3610 

gac ctt ggc cag gaa gtg gcc ctg aat gct aac act aag aac cag aag    11018 
Asp Leu Gly Gln Glu Val Ala Leu Asn Ala Asn Thr Lys Asn Gln Lys 
3615                3620                3625                3630 

atc aga tgg aaa aat gaa gtc cgg att cat tct ggg tct ttc cag agc    11066 
Ile Arg Trp Lys Asn Glu Val Arg Ile His Ser Gly Ser Phe Gln Ser 
                3635                3640                3645 

cag gtc gag ctt tcc aat gac caa gaa aag gca cac ctt gac att gca    11114 
Gln Val Glu Leu Ser Asn Asp Gln Glu Lys Ala His Leu Asp Ile Ala 
            3650                3655                3660 

gga tcc tta gaa gga cac cta agg ttc ctc aaa aat atc atc cta cca    11162 
Gly Ser Leu Glu Gly His Leu Arg Phe Leu Lys Asn Ile Ile Leu Pro 
        3665                3670                3675 

gtc tat gac aag agc tta tgg gat ttc cta aag ctg gat gta acc acc    11210 
Val Tyr Asp Lys Ser Leu Trp Asp Phe Leu Lys Leu Asp Val Thr Thr 
    3680                3685                3690 

agc att ggt agg aga cag cat ctt cgt gtt tca act gcc ttt gtg tac    11258 
Ser Ile Gly Arg Arg Gln His Leu Arg Val Ser Thr Ala Phe Val Tyr 
3695                3700                3705                3710 

acc aaa aac ccc aat ggc tat tca ttc tcc atc cct gta aaa gtt ttg    11306 
Thr Lys Asn Pro Asn Gly Tyr Ser Phe Ser Ile Pro Val Lys Val Leu 
                3715                3720                3725 

gct gat aaa ttc att act cct ggg ctg aaa cta aat gat cta aat tca    11354 
Ala Asp Lys Phe Ile Thr Pro Gly Leu Lys Leu Asn Asp Leu Asn Ser 
            3730                3735                3740 

gtt ctt gtc atg cct acg ttc cat gtc cca ttt aca gat ctt cag gtt    11402 
Val Leu Val Met Pro Thr Phe His Val Pro Phe Thr Asp Leu Gln Val 
        3745                3750                3755 

cca tcg tgc aaa ctt gac ttc aga gaa ata caa atc tat aag aag ctg    11450 
Pro Ser Cys Lys Leu Asp Phe Arg Glu Ile Gln Ile Tyr Lys Lys Leu 
    3760                3765                3770 

aga act tca tca ttt gcc ctc aac cta cca aca ctc ccc gag gta aaa    11498 
Arg Thr Ser Ser Phe Ala Leu Asn Leu Pro Thr Leu Pro Glu Val Lys 
3775                3780                3785                3790 

ttc cct gaa gtt gat gtg tta aca aaa tat tct caa cca gaa gac tcc    11546 
Phe Pro Glu Val Asp Val Leu Thr Lys Tyr Ser Gln Pro Glu Asp Ser 
                3795                3800                3805 

ttg att ccc ttt ttt gag ata acc gtg cct gaa tct cag tta act gtg    11594 
Leu Ile Pro Phe Phe Glu Ile Thr Val Pro Glu Ser Gln Leu Thr Val 
            3810                3815                3820 

tcc cag ttc acg ctt cca aaa agt gtt tca gat ggc att gct gct ttg    11642 
Ser Gln Phe Thr Leu Pro Lys Ser Val Ser Asp Gly Ile Ala Ala Leu 
        3825                3830                3835 

gat cta aat gca gta gcc aac aag atc gca gac ttt gag ttg ccc acc    11690 
Asp Leu Asn Ala Val Ala Asn Lys Ile Ala Asp Phe Glu Leu Pro Thr 
    3840                3845                3850 

atc atc gtg cct gag cag acc att gag att ccc tcc att aag ttc tct    11738 
Ile Ile Val Pro Glu Gln Thr Ile Glu Ile Pro Ser Ile Lys Phe Ser 
3855                3860                3865                3870 

gta cct gct gga att gtc att cct tcc ttt caa gca ctg act gca cgc    11786 
Val Pro Ala Gly Ile Val Ile Pro Ser Phe Gln Ala Leu Thr Ala Arg 
                3875                3880                3885 

ttt gag gta gac tct ccc gtg tat aat gcc act tgg agt gcc agt ttg    11834 
Phe Glu Val Asp Ser Pro Val Tyr Asn Ala Thr Trp Ser Ala Ser Leu 
            3890                3895                3900 

aaa aac aaa gca gat tat gtt gaa aca gtc ctg gat tcc aca tgc agc    11882 
Lys Asn Lys Ala Asp Tyr Val Glu Thr Val Leu Asp Ser Thr Cys Ser 
        3905                3910                3915 

tca acc gta cag ttc cta gaa tat gaa cta aat gtt ttg gga aca cac    11930 
Ser Thr Val Gln Phe Leu Glu Tyr Glu Leu Asn Val Leu Gly Thr His 
    3920                3925                3930 

aaa atc gaa gat ggt acg tta gcc tct aag act aaa gga aca ctt gca    11978 
Lys Ile Glu Asp Gly Thr Leu Ala Ser Lys Thr Lys Gly Thr Leu Ala 
3935                3940                3945                3950 

cac cgt gac ttc agt gca gaa tat gaa gaa gat ggc aaa ttt gaa gga    12026 
His Arg Asp Phe Ser Ala Glu Tyr Glu Glu Asp Gly Lys Phe Glu Gly 
                3955                3960                3965 

ctt cag gaa tgg gaa gga aaa gcg cac ctc aat atc aaa agc cca gcg    12074 
Leu Gln Glu Trp Glu Gly Lys Ala His Leu Asn Ile Lys Ser Pro Ala 
            3970                3975                3980 

ttc acc gat ctc cat ctg cgc tac cag aaa gac aag aaa ggc atc tcc    12122 
Phe Thr Asp Leu His Leu Arg Tyr Gln Lys Asp Lys Lys Gly Ile Ser 
        3985                3990                3995 

acc tca gca gcc tcc cca gcc gta ggc acc gtg ggc atg gat atg gat    12170 
Thr Ser Ala Ala Ser Pro Ala Val Gly Thr Val Gly Met Asp Met Asp 
    4000                4005                4010 

gaa gat gac gac ttt tct aaa tgg aac ttc tac tac agc cct cag tcc    12218 
Glu Asp Asp Asp Phe Ser Lys Trp Asn Phe Tyr Tyr Ser Pro Gln Ser 
4015                4020                4025                4030 

tct cca gat aaa aaa ctc acc ata ttc aaa act gag ttg agg gtc cgg    12266 
Ser Pro Asp Lys Lys Leu Thr Ile Phe Lys Thr Glu Leu Arg Val Arg 
                4035                4040                4045 

gaa tct gat gag gaa act cag atc aaa gtt aat tgg gaa gaa gag gca    12314 
Glu Ser Asp Glu Glu Thr Gln Ile Lys Val Asn Trp Glu Glu Glu Ala 
            4050                4055                4060 

gct tct ggc ttg cta acc tct ctg aaa gac aac gtg ccc aag gcc aca    12362 
Ala Ser Gly Leu Leu Thr Ser Leu Lys Asp Asn Val Pro Lys Ala Thr 
        4065                4070                4075 

ggg gtc ctt tat gat tat gtc aac aag tac cac tgg gaa cac aca ggg    12410 
Gly Val Leu Tyr Asp Tyr Val Asn Lys Tyr His Trp Glu His Thr Gly 
    4080                4085                4090 

ctc acc ctg aga gaa gtg tct tca aag ctg aga aga aat ctg cag aac    12458 
Leu Thr Leu Arg Glu Val Ser Ser Lys Leu Arg Arg Asn Leu Gln Asn 
4095                4100                4105                4110 

aat gct gag tgg gtt tat caa ggg gcc att agg caa att gat gat atc    12506 
Asn Ala Glu Trp Val Tyr Gln Gly Ala Ile Arg Gln Ile Asp Asp Ile 
                4115                4120                4125 

gac gtg agg ttc cag aaa gca gcc agt ggc acc act ggg acc tac caa    12554 
Asp Val Arg Phe Gln Lys Ala Ala Ser Gly Thr Thr Gly Thr Tyr Gln 
            4130                4135                4140 

gag tgg aag gac aag gcc cag aat ctg tac cag gaa ctg ttg act cag    12602 
Glu Trp Lys Asp Lys Ala Gln Asn Leu Tyr Gln Glu Leu Leu Thr Gln 
        4145                4150                4155 

gaa ggc caa gcc agt ttc cag gga ctc aag gat aac gtg ttt gat ggc    12650 
Glu Gly Gln Ala Ser Phe Gln Gly Leu Lys Asp Asn Val Phe Asp Gly 
    4160                4165                4170 

ttg gta cga gtt act caa aaa ttc cat atg aaa gtc aag cat ctg att    12698 
Leu Val Arg Val Thr Gln Lys Phe His Met Lys Val Lys His Leu Ile 
4175                4180                4185                4190 

gac tca ctc att gat ttt ctg aac ttc ccc aga ttc cag ttt ccg ggg    12746 
Asp Ser Leu Ile Asp Phe Leu Asn Phe Pro Arg Phe Gln Phe Pro Gly 
                4195                4200                4205 

aaa cct ggg ata tac act agg gag gaa ctt tgc act atg ttc ata agg    12794 
Lys Pro Gly Ile Tyr Thr Arg Glu Glu Leu Cys Thr Met Phe Ile Arg 
            4210                4215                4220 

gag gta ggg acg gta ctg tcc cag gta tat tcg aaa gtc cat aat ggt    12842 
Glu Val Gly Thr Val Leu Ser Gln Val Tyr Ser Lys Val His Asn Gly 
        4225                4230                4235 

tca gaa ata ctg ttt tcc tat ttc caa gac cta gtg att aca ctt cct    12890 
Ser Glu Ile Leu Phe Ser Tyr Phe Gln Asp Leu Val Ile Thr Leu Pro 
    4240                4245                4250 

ttc gag tta agg aaa cat aaa cta ata gat gta atc tcg atg tat agg    12938 
Phe Glu Leu Arg Lys His Lys Leu Ile Asp Val Ile Ser Met Tyr Arg 
4255                4260                4265                4270 

gaa ctg ttg aaa gat tta tca aaa gaa gcc caa gag gta ttt aaa gcc    12986 
Glu Leu Leu Lys Asp Leu Ser Lys Glu Ala Gln Glu Val Phe Lys Ala 
                4275                4280                4285 

att cag tct ctc aag acc aca gag gtg cta cgt aat ctt cag gac ctt    13034 
Ile Gln Ser Leu Lys Thr Thr Glu Val Leu Arg Asn Leu Gln Asp Leu 
            4290                4295                4300 

tta caa ttc att ttc caa cta ata gaa gat aac att aaa cag ctg aaa    13082 
Leu Gln Phe Ile Phe Gln Leu Ile Glu Asp Asn Ile Lys Gln Leu Lys 
        4305                4310                4315 

gag atg aaa ttt act tat ctt att aat tat atc caa gat gag atc aac    13130 
Glu Met Lys Phe Thr Tyr Leu Ile Asn Tyr Ile Gln Asp Glu Ile Asn 
    4320                4325                4330 

aca atc ttc aat gat tat atc cca tat gtt ttt aaa ttg ttg aaa gaa    13178 
Thr Ile Phe Asn Asp Tyr Ile Pro Tyr Val Phe Lys Leu Leu Lys Glu 
4335                4340                4345                4350 

aac cta tgc ctt aat ctt cat aag ttc aat gaa ttt att caa aac gag    13226 
Asn Leu Cys Leu Asn Leu His Lys Phe Asn Glu Phe Ile Gln Asn Glu 
                4355                4360                4365 

ctt cag gaa gct tct caa gag tta cag cag atc cat caa tac att atg    13274 
Leu Gln Glu Ala Ser Gln Glu Leu Gln Gln Ile His Gln Tyr Ile Met 
            4370                4375                4380 

gcc ctt cgt gaa gaa tat ttt gat cca agt ata gtt ggc tgg aca gtg    13322 
Ala Leu Arg Glu Glu Tyr Phe Asp Pro Ser Ile Val Gly Trp Thr Val 
        4385                4390                4395 

aaa tat tat gaa ctt gaa gaa aag ata gtc agt ctg atc aag aac ctg    13370 
Lys Tyr Tyr Glu Leu Glu Glu Lys Ile Val Ser Leu Ile Lys Asn Leu 
    4400                4405                4410 

tta gtt gct ctt aag gac ttc cat tct gaa tat att gtc agt gcc tct    13418 
Leu Val Ala Leu Lys Asp Phe His Ser Glu Tyr Ile Val Ser Ala Ser 
4415                4420                4425                4430 

aac ttt act tcc caa ctc tca agt caa gtt gag caa ttt ctg cac aga    13466 
Asn Phe Thr Ser Gln Leu Ser Ser Gln Val Glu Gln Phe Leu His Arg 
                4435                4440                4445 

aat att cag gaa tat ctt agc atc ctt acc gat cca gat gga aaa ggg    13514 
Asn Ile Gln Glu Tyr Leu Ser Ile Leu Thr Asp Pro Asp Gly Lys Gly 
            4450                4455                4460 

aaa gag aag att gca gag ctt tct gcc act gct cag gaa ata att aaa    13562 
Lys Glu Lys Ile Ala Glu Leu Ser Ala Thr Ala Gln Glu Ile Ile Lys 
        4465                4470                4475 

agc cag gcc att gcg acg aag aaa ata att tct gat tac cac cag cag    13610 
Ser Gln Ala Ile Ala Thr Lys Lys Ile Ile Ser Asp Tyr His Gln Gln 
    4480                4485                4490 

ttt aga tat aaa ctg caa gat ttt tca gac caa ctc tct gat tac tat    13658 
Phe Arg Tyr Lys Leu Gln Asp Phe Ser Asp Gln Leu Ser Asp Tyr Tyr 
4495                4500                4505                4510 

gaa aaa ttt att gct gaa tcc aaa aga ttg att gac ctg tcc att caa    13706 
Glu Lys Phe Ile Ala Glu Ser Lys Arg Leu Ile Asp Leu Ser Ile Gln 
                4515                4520                4525 

aac tac cac aca ttt ctg ata tac atc acg gag tta ctg aaa aag ctg    13754 
Asn Tyr His Thr Phe Leu Ile Tyr Ile Thr Glu Leu Leu Lys Lys Leu 
            4530                4535                4540 

caa tca acc aca gtc atg aac ccc tac atg aag ctt gct cca gga gaa   13802 
Gln Ser Thr Thr Val Met Asn Pro Tyr Met Lys Leu Ala Pro Gly Glu 
        4545                4550                4555 

ctt act atc atc ctc taa ttttttaaaa gaaatcttca tttattcttc           13850 
Leu Thr Ile Ile Leu  * 
    4560 

ttttccaatt gaactttcac atagcacaga aaaaattcaa actgcctata ttgataaaac  13910 

catacagtga gccagccttg cagtaggcag tagactataa gcagaagcac atatgaactg  13970 

gacctgcacc aaagctggca ccagggctcg gaaggtctct gaactcagaa ggatggcatt  14030 

ttttgcaagt taaagaaaat caggatctga gttattttgc taaacttggg ggaggaggaa  14090 

caaataaatg gagtctttat tgtgtatcat a                                 14121 

 
           
             546  
             21  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            546 

tgctaaaggc acatatggcc t                                               21 

 
           
             547  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            547 

ctcaggttgg actctccatt gag                                             23 

 
           
             548  
             28  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            548 

cttgtcagag ggatcctaac actggccg                                        28 

 
           
             549  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            549 

ccgcaggtcc cggtgggaat                                                 20 

 
           
             550  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            550 

accgagaagg gcactcagcc                                                 20 

 
           
             551  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            551 

gcctcggcct cgcggccctg                                                 20 

 
           
             552  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            552 

tccatcgcca gctgcggtgg                                                 20 

 
           
             553  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            553 

cagcgccagc agcgccagca                                                 20 

 
           
             554  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            554 

gcccgccagc agcagcagca                                                 20 

 
           
             555  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            555 

cttgaatcag cagtcccagg                                                 20 

 
           
             556  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            556 

cttcagcaag gctttgccct                                                 20 

 
           
             557  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            557 

tttctgttgc cacattgccc                                                 20 

 
           
             558  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            558 

ggaagaggtg ttgctccttg                                                 20 

 
           
             559  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            559 

tgtgctacca tcccatactt                                                 20 

 
           
             560  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            560 

tcaaatgcga ggcccatctt                                                 20 

 
           
             561  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            561 

ggacacctca atcagctgtg                                                 20 

 
           
             562  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            562 

tcagggccac caggtaggtg                                                 20 

 
           
             563  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            563 

gtaatcttca tccccagtgc                                                 20 

 
           
             564  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            564 

tgctccatgg tttggcccat                                                 20 

 
           
             565  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            565 

gcagccagtc gcttatctcc                                                 20 

 
           
             566  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            566 

gtatagccaa agtggtccac                                                 20 

 
           
             567  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            567 

cccaggagct ggaggtcatg                                                 20 

 
           
             568  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            568 

ttgagccctt cctgatgacc                                                 20 

 
           
             569  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            569 

atctggaccc cactcctagc                                                 20 

 
           
             570  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            570 

cagacccgac tcgtggaaga                                                 20 

 
           
             571  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            571 

gccctcagta gattcatcat                                                 20 

 
           
             572  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            572 

gccatgccac cctcttggaa                                                 20 

 
           
             573  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            573 

aacccacgtg ccggaaagtc                                                 20 

 
           
             574  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            574 

actcccagat gccttctgaa                                                 20 

 
           
             575  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            575 

atgtggtaac gagcccgaag                                                 20 

 
           
             576  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            576 

ggcgtagaga cccatcacat                                                 20 

 
           
             577  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            577 

gtgttaggat ccctctgaca                                                 20 

 
           
             578  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            578 

cccagtgata gctctgtgag                                                 20 

 
           
             579  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            579 

atttcagcat atgagcccat                                                 20 

 
           
             580  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            580 

ccctgaacct tagcaacagt                                                 20 

 
           
             581  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            581 

gctgaagcca gcccagcgat                                                 20 

 
           
             582  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            582 

acagctgccc agtatgttct                                                 20 

 
           
             583  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            583 

cccaataaga tttataacaa                                                 20 

 
           
             584  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            584 

tggcctacca gagacaggta                                                 20 

 
           
             585  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            585 

tcatacgttt agcccaatct                                                 20 

 
           
             586  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            586 

gcatggtccc aaggatggtc                                                 20 

 
           
             587  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            587 

agtgatggaa gctgcgatac                                                 20 

 
           
             588  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            588 

atgagcatca tgcctcccag                                                 20 

 
           
             589  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            589 

gaacacatag ccgaatgccg                                                 20 

 
           
             590  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            590 

gtggtgccct ctaatttgta                                                 20 

 
           
             591  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            591 

cccgagaaag aaccgaaccc                                                 20 

 
           
             592  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            592 

tgccctgcag cttcactgaa                                                 20 

 
           
             593  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            593 

gaaatcccat aagctcttgt                                                 20 

 
           
             594  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            594 

agaagctgcc tcttcttccc                                                 20 

 
           
             595  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            595 

tcagggtgag ccctgtgtgt                                                 20 

 
           
             596  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            596 

ctaatggccc cttgataaac                                                 20 

 
           
             597  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            597 

acgttatcct tgagtccctg                                                 20 

 
           
             598  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            598 

tatatcccag gtttccccgg                                                 20 

 
           
             599  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            599 

acctgggaca gtaccgtccc                                                 20 

 
           
             600  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            600 

ctgcctactg caaggctggc                                                 20 

 
           
             601  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            601 

agagaccttc cgagccctgg                                                 20 

 
           
             602  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            602 

atgatacaca ataaagactc                                                 20 

 
           
             603  
             2354  
             DNA  
             Mus musculus  
             
               allele  
               (0)...(0)  
             
           
            603 

gaattccaac ttcctcacct ctcacataca attgaaatac ctgcttttgg caaactgcat     60 

agcatcctta agatccaatc tcctctcttt atattagatg ctaatgccaa catacagaat    120 

gtaacaactt cagggaacaa agcagagatt gtggcttctg tcactgctaa aggagagtcc    180 

caatttgaag ctctcaattt tgattttcaa gcacaagctc aattcctgga gttaaatcct    240 

catcctccag tcctgaagga atccatgaac ttctccagta agcatgtgag aatggagcat    300 

gagggtgaga tagtatttga tggaaaggcc attgagggga aatcagacac agtcgcaagt    360 

ttacacacag agaaaaatga agtagagttt aataatggta tgactgtcaa agtaaacaat    420 

cagctcaccc ttgacagtca cacaaagtac ttccacaagt tgagtgttcc taggctggac    480 

ttctccagta aggcttctct taataatgaa atcaagacac tattagaagc tggacatgtg    540 

gcattgacat cttcagggac agggtcatgg aactgggcct gtcccaactt ctcggatgaa    600 

ggcatacatt cgtcccaaat tagctttact gtggatggtc ccattgcttt tgttggacta    660 

tccaataaca taaatggcaa acacttacgg gtcatccaaa aactgactta tgaatctggc    720 

ttcctcaact attctaagtt tgaagttgag tcaaaagttg aatctcagca cgtgggctcc    780 

agcattctaa cagccaatgg tcgggcactg ctcaaggacg caaaggcaga aatgactggt    840 

gagcacaatg ccaacttaaa tggaaaagtt attggaactt tgaaaaattc tctcttcttt    900 

tcagcacaac catttgagat tactgcatcc acaaataatg aaggaaattt gaaagtgggt    960 

tttccactaa agctgactgg gaaaatagac ttcctgaata actatgcatt gtttctgagt   1020 

ccccgtgccc aacaagcaag ctggcaagcg agtaccagat tcaatcagta caaatacaat   1080 

caaaactttt ctgctataaa caatgaacac aacatagaag ccagtatagg aatgaatgga   1140 

gatgccaacc tggatttctt aaacatacct ttaacaattc ctgaaattaa cttgccttac   1200 

acggagttca aaactccctt actgaaggat ttctccatat gggaagaaac aggcttgaaa   1260 

gaatttttga agacaacaaa gcaatcattt gatttgagtg taaaggctca atataaaaag   1320 

aacagtgaca agcattccat tgttgtccct ctgggtatgt tttatgaatt tattctcaac   1380 

aatgtcaatt cgtgggacag aaaatttgag aaagtcagaa acaatgcttt acattttctt   1440 

accacctcct ataatgaagc aaaaattaag gttgataagt acaaaactga aaattccctt   1500 

aatcagccct ctgggacctt tcaaaatcat ggctacacta tcccagttgt caacattgaa   1560 

gtatctccat ttgctgtaga gacactggct tccaggcatg tgatccccac agcaataagc   1620 

accccaagtg tcacaatccc tggtcctaac atcatggtgc cttcatacaa gttagtgctg   1680 

ccacccctgg agttgccagt tttccatggt cctgggaatc tattcaagtt tttcctccca   1740 

gatttcaagg gattcaacac tattgacaat atttatattc cagccatggg caactttacc   1800 

tatgactttt cttttaaatc aagtgtcatc acactgaata ccaatgctgg actttataac   1860 

caatcagata tcgttgccca tttcctttct tcctcttcat ttgtcactga cgccctgcag   1920 

tacaaattag agggaacatc acgtctgatg cgaaaaaggg gattgaaact agccacagct   1980 

gtctctctaa ctaacaaatt tgtaaagggc agtcatgaca gcaccattag tttaaccaag   2040 

aaaaacatgg aagcatcagt gagaacaact gccaacctcc atgctcccat attctcaatg   2100 

aacttcaagc aggaacttaa tggaaatacc aagtcaaaac ccactgtttc atcatccatt   2160 

gaactaaact atgacttcaa ttcctcaaag ctgcactcta ctgcaacagg aggcattgat   2220 

cacaagttca gcttagaaag tctcacttcc tacttttcca ttgagtcatt caccaaagga   2280 

aatatcaaga gttccttcct ttctcaggaa tattcaggaa gtgttgccaa tgaagccaat   2340 

gtatatctga attc                                                     2354 

 
           
             604  
             19  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            604 

cgtgggctcc agcattcta                                                  19 

 
           
             605  
             21  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            605 

agtcatttct gcctttgcgt c                                               21 

 
           
             606  
             22  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            606 

ccaatggtcg ggcactgctc aa                                              22 

 
           
             607  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            607 

attgtatgtg agaggtgagg                                                 20 

 
           
             608  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            608 

gaggagattg gatcttaagg                                                 20 

 
           
             609  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            609 

cttcaaattg ggactctcct                                                 20 

 
           
             610  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            610 

tccaggaatt gagcttgtgc                                                 20 

 
           
             611  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            611 

ttcaggactg gaggatgagg                                                 20 

 
           
             612  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            612 

tctcaccctc atgctccatt                                                 20 

 
           
             613  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            613 

tgactgtcaa gggtgagctg                                                 20 

 
           
             614  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            614 

gtccagccta ggaacactca                                                 20 

 
           
             615  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            615 

atgtcaatgc cacatgtcca                                                 20 

 
           
             616  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            616 

ttcatccgag aagttgggac                                                 20 

 
           
             617  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            617 

atttgggacg aatgtatgcc                                                 20 

 
           
             618  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            618 

agttgaggaa gccagattca                                                 20 

 
           
             619  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            619 

ttcccagtca gctttagtgg                                                 20 

 
           
             620  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            620 

agcttgcttg ttgggcacgg                                                 20 

 
           
             621  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            621 

cctatactgg cttctatgtt                                                 20 

 
           
             622  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            622 

tgaactccgt gtaaggcaag                                                 20 

 
           
             623  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            623 

gagaaatcct tcagtaaggg                                                 20 

 
           
             624  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            624 

caatggaatg cttgtcactg                                                 20 

 
           
             625  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            625 

gcttcattat aggaggtggt                                                 20 

 
           
             626  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            626 

acaactggga tagtgtagcc                                                 20 

 
           
             627  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            627 

gttaggacca gggattgtga                                                 20 

 
           
             628  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            628 

accatggaaa actggcaact                                                 20 

 
           
             629  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            629 

tgggaggaaa aacttgaata                                                 20 

 
           
             630  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            630 

tgggcaacga tatctgattg                                                 20 

 
           
             631  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            631 

ctgcagggcg tcagtgacaa                                                 20 

 
           
             632  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            632 

gcatcagacg tgatgttccc                                                 20 

 
           
             633  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            633 

cttggttaaa ctaatggtgc                                                 20 

 
           
             634  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            634 

atgggagcat ggaggttggc                                                 20 

 
           
             635  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            635 

aatggatgat gaaacagtgg                                                 20 

 
           
             636  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            636 

atcaatgcct cctgttgcag                                                 20 

 
           
             637  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            637 

ggaagtgaga ctttctaagc                                                 20 

 
           
             638  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            638 

aggaaggaac tcttgatatt                                                 20 

 
           
             639  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            639 

attggcttca ttggcaacac                                                 20 

 
           
             640  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            640 

aggtgaggaa gttggaattc                                                 20 

 
           
             641  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            641 

ttgttccctg aagttgttac                                                 20 

 
           
             642  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            642 

gttcatggat tccttcagga                                                 20 

 
           
             643  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            643 

atgctccatt ctcacatgct                                                 20 

 
           
             644  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            644 

tgcgactgtg tctgatttcc                                                 20 

 
           
             645  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            645 

gtccctgaag atgtcaatgc                                                 20 

 
           
             646  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            646 

aggcccagtt ccatgaccct                                                 20 

 
           
             647  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            647 

ggagcccacg tgctgagatt                                                 20 

 
           
             648  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            648 

cgtccttgag cagtgcccga                                                 20 

 
           
             649  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            649 

cccatatgga gaaatccttc                                                 20 

 
           
             650  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            650 

catgcctgga agccagtgtc                                                 20 

 
           
             651  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            651 

gtgttgaatc ccttgaaatc                                                 20 

 
           
             652  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            652 

ggtaaagttg cccatggctg                                                 20 

 
           
             653  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            653 

gttataaagt ccagcattgg                                                 20 

 
           
             654  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            654 

catcagacgt gatgttccct                                                 20 

 
           
             655  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            655 

tggctagttt caatcccctt                                                 20 

 
           
             656  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            656 

ctgtcatgac tgccctttac                                                 20 

 
           
             657  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            657 

gcttgaagtt cattgagaat                                                 20 

 
           
             658  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            658 

ttcctgagaa aggaaggaac                                                 20 

 
           
             659  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            659 

tcagatatac attggcttca                                                 20 

 
           
             660  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            660 

tcgatctcct tttatgcccg                                                 20 

 
           
             661  
             1105  
             DNA  
             Homo sapiens  
             
               CDS  
               (141)...(728)  
             
           
            661 

gggcgggtag tcgaccgtgt ccgcgcgcct gggagacgct gcctcggccc ggacgcgccc     60 

gcgcccccgc ggctggaggg tggtcgccac tgggacactg tgaaccagga gtgagtcgga    120 

gctgccgcgc tgcccaggcc atg gac tgt gag gtc aac aac ggt tcc agc ctc    173 
                       Met Asp Cys Glu Val Asn Asn Gly Ser Ser Leu 
                        1               5                   10 

agg gat gag tgc atc aca aac cta ctg gtg ttt ggc ttc ctc caa agc      221 
Arg Asp Glu Cys Ile Thr Asn Leu Leu Val Phe Gly Phe Leu Gln Ser 
             15                  20                  25 

tgt tct gac aac agc ttc cgc aga gag ctg gac gca ctg ggc cac gag      269 
Cys Ser Asp Asn Ser Phe Arg Arg Glu Leu Asp Ala Leu Gly His Glu 
         30                  35                  40 

ctg cca gtg ctg gct ccc cag tgg gag ggc tac gat gag ctg cag act      317 
Leu Pro Val Leu Ala Pro Gln Trp Glu Gly Tyr Asp Glu Leu Gln Thr 
     45                  50                  55 

gat ggc aac cgc agc agc cac tcc cgc ttg gga aga ata gag gca gat      365 
Asp Gly Asn Arg Ser Ser His Ser Arg Leu Gly Arg Ile Glu Ala Asp 
 60                  65                  70                  75 

tct gaa agt caa gaa gac atc atc cgg aat att gcc agg cac ctc gcc      413 
Ser Glu Ser Gln Glu Asp Ile Ile Arg Asn Ile Ala Arg His Leu Ala 
                 80                  85                  90 

cag gtc ggg gac agc atg gac cgt agc atc cct ccg ggc ctg gtg aac      461 
Gln Val Gly Asp Ser Met Asp Arg Ser Ile Pro Pro Gly Leu Val Asn 
             95                 100                 105 

ggc ctg gcc ctg cag ctc agg aac acc agc cgg tcg gag gag gac cgg      509 
Gly Leu Ala Leu Gln Leu Arg Asn Thr Ser Arg Ser Glu Glu Asp Arg 
        110                 115                 120 

aac agg gac ctg gcc act gcc ctg gag cag ctg ctg cag gcc tac cct      557 
Asn Arg Asp Leu Ala Thr Ala Leu Glu Gln Leu Leu Gln Ala Tyr Pro 
    125                 130                 135 

aga gac atg gag aag gag aag acc atg ctg gtg ctg gcc ctg ctg ctg      605 
Arg Asp Met Glu Lys Glu Lys Thr Met Leu Val Leu Ala Leu Leu Leu 
140                 145                 150                 155 

gcc aag aag gtg gcc agt cac acg ccg tcc ttg ctc cgt gat gtc ttt      653 
Ala Lys Lys Val Ala Ser His Thr Pro Ser Leu Leu Arg Asp Val Phe 
                160                 165                 170 

cac aca aca gtg aat ttt att aac cag aac cta cgc acc tac gtg agg      701 
His Thr Thr Val Asn Phe Ile Asn Gln Asn Leu Arg Thr Tyr Val Arg 
            175                 180                 185 

agc tta gcc aga aat ggg atg gac tga acggacagtt ccagaagtgt            748 
Ser Leu Ala Arg Asn Gly Met Asp  * 
        190                 195 

gactggctaa agcttgatgt ggtcacagct gtatagctgc ttccagtgta gacggagccc    808 

tggcatgtca acagcgttcc tagagaagac aggctggaag atagctgtga cttctatttt    868 

aaagacaatg ttaaacttat aacccacttt aaaatatcta cattaatata cttgaatgaa    928 

aatgtccatt tacacgtatt tgaatggcct tcatatcatc cacacatgaa tctgcacatc    988 

tgtaaatcta cacacggtgc ctttatttcc actgtgcagg ttcccactta aaaattaaat   1048 

tggaaagcag gtttcaagga agtagaaaca aaatacaatt tttttggtaa aaaaaaa      1105 

 
           
             662  
             24  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            662 

agaagacatc atccggaata ttgc                                            24 

 
           
             663  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            663 

ggagggatgc tacggtccat                                                 20 

 
           
             664  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            664 

aggcacctcg cccaggtcgg                                                 20 

 
           
             665  
             18000  
             DNA  
             Homo sapiens  
             
               CDS  
               (14031)...(14243)  
               Antisense Oligonucleotide  
             
           
            665 

cctgggtatc caagtcgccc tggcagagaa acactgcatg agacacggcg ttagggtctg     60 

gtgggagact caccacagtg ccaaggtggc tgcagtttgc ttgtgacatg ggcgtgtatc    120 

tgagtgtgaa ggaagctggt ttttgtgagc tgcctcccga gctcagaggt gacagtgggc    180 

actttcccca cagagacccc tgaagttgtt ccttggagaa caaagtggtg aggggcgggg    240 

attccagacc ttgaggcaga agctagggtc tggtccactg ttctgtggac tgggcagtgg    300 

ccctgggagg tgccgtggcc tctgtggcct gtttcctggg gtggggtctg tcttgcgctt    360 

tgtctcttgt gggtgcagac tccccttcct ctgctgtgga gccggcagat ggccccggag    420 

ccagatcctg gtgcctccct gtccacatgc agctcagtca tttgctcttg gtcccttcct    480 

atgaaatgca cggccacaca cagccagggt ttctcctggg ctccccagag ggagagtagg    540 

gtgcagcctg caacagtgca gggtccccag gcctgtgtga gcccccaggt ggggaggtgg    600 

gtgatgcgca tgtcagtgct acctcctgcc acctcctctc tgcctgggca caggctttct    660 

cctctgtttg ctttttattt cctatgtatt caggaaccat gtgaaattgc caatgcttgg    720 

ttttgtccta caaaatggcc atttcatttg gttcaacctg atattgtgtc tacacacaca    780 

cacgcacaca cacacacaca caggcaaata ctttttaaaa caggattatt ctattcacag    840 

tgttctgtag aaatttgtgt tcagtctttt tttttttttt tgagacggag tctcgctctg    900 

tcgcccaggt cggactgcgg actgcagtgg cgcaatctcg gctcactgca agctccgctt    960 

cccgggttca cgccattctc ctgcctcagc ctcccgagta gctgggacta caggcgcccg   1020 

ccaccgcgcc cggctaattt tttgtatttt tagtagagac ggggtttcac cttgttagcc   1080 

aggatggtct cgatctcctg acctcatgat ccacccgcct cggcctccca aagtgctggg   1140 

attacaggcg tgagccaccg cgcccggcct cagtcttttt aagacagctt actgtactga   1200 

tgccgcacag atcttttttt tttttcgaga cagggtttca ctctcgccca ggctggagtg   1260 

cagtggtgca atctccgctc actgcagcct ccacctcctg ggtgtaagtg atcctcctgc   1320 

ttcagccccc caagtagctg ggcccacagg gcttgcatca ccacacctgg ctaattttgt   1380 

atttttgtag agatggggtt tcaccatgtt ggccagactg gtcattcttt ttgagatgga   1440 

gtctcgctct gtcgcccagg ctggagtgca gtggcgtgat ctcggcttac tgcaacctct   1500 

ccctcccaag ttcatgccat tctcctgcct cagcctcccg agtagctggg actacaggcg   1560 

cccgccacca cgcccggcta attttttgta tttttagtag agacagggtt tcaccgcatt   1620 

agccagggtg gtctcgatct cctgacctca tgatccaccc gcctcggcct cccaaagtgc   1680 

tgggattaca ggcatgagct actgcgtcca gccggaagat ttaatttttt aattgtcaaa   1740 

tccattctct ctctctataa acattttaca ttttatgata ataaaataat ttgtgagccc   1800 

acggccccgt ttccctgatg cctgaggtct tcctggggcg gcatgggagg gctgaattca   1860 

ggtgcggggt cggccccagg gcactgagcg cctgggtgag tatctggaat gaggaaaaca   1920 

aagcttggct cccgccaagg agaaagaaac tcaggatgcg gggctcaggc caggacctcg   1980 

gctcagccgc catttctgga gcacaggcca gcttcgtcgt cctcccgagg ggtcctgacc   2040 

agggcttccc aggagcggcc gcccactctg tgtgtccctt tccaggtcgc cactgggaca   2100 

ctgtgaacca ggagtgagtc ggagctgccg cgctgcccag gccatggact gtgaggtcag   2160 

aggccagatc ccctgcgggt gccttgtggg gggcggggtc gaggggtaag ggcctgcgtg   2220 

tcccccacca cgcatccctg agggctgagg ctgagcccgc ctggccctta ccacagctcg   2280 

gcacagacga accccgccca gccccttcac tgaagcaggc gggagccggg aagtcctacc   2340 

tttccctgtc ctgcgccttc ctcgcactcc gcttgtggtg cagcccctcc acaccgcgcc   2400 

tggggctaac tgcaagggcg agggggcttt gggtttaaga ccatttaaca gccataggct   2460 

gtgggtccca gcactttggg aggccaaggc aggaggattc cttgaggcca ggaggtcgag   2520 

gctacagtga gctgtgattg tgccactgca ctgcagccct gtccaaacaa acacgaaaga   2580 

gatttaagaa gaagaaaggg ggcattagat aagcacttca tataattctc tcaactgtaa   2640 

aagcaagaca atacttacct tgtctaacca atgccattgc tatgaggagc aaataaatca   2700 

ataaaggtca aataaaagta ctgtaaactg taaggtgttt caaaaatttt ttaacccact   2760 

ggatttaaat ttcccttcat agctgggcga ggtggcttag gcacataatc ccagtgactt   2820 

gggaggcaga agcgagagga ttgcttgaag ccaggagttt gattgagaca aacctgggca   2880 

acatagtaag accccgtctt tataaagata aaagcggtgg agttctggga ggggagcccg   2940 

gagcccccgc cttcagcagg acgctccctg gatgcttcct tgtctctcct tccctttaaa   3000 

tggtctgggg agagaaaaat cacagcacac gggtgctctc tcccacccgc tgcatcacat   3060 

cctcctcccc tccctcctgc cgaattctgc agcctctggg cgcctcacgc tgtcctggca   3120 

gcctctggga aggcatctgc gaagtctaat gccttggcac ttagtgactg tgtcgcagtt   3180 

cctgagcatg gagagcaccc ggcacccagg aggttctcaa gctgccccta ctgggggtcc   3240 

tttccaaagg tggggacggt gtggatttca gcgtggtggc tggagggctg aggcagtggc   3300 

tcgagtttga tgttagttac ataaacagag gagattgcag gagctccccc ggccctgatc   3360 

caggcttgtt gtcagtgtcc aaaagaccac tctgggtgcc actgtccctt cccacctgcc   3420 

gctgctgttc cggcttcgcg ctctggcggc ctccgcaggt agaacaccac cgtcacccgc   3480 

gcagcgccct gactcgccgg aggaggcgcc tgccctcccg cccgcctctc cccggccccc   3540 

tcagtgaggg agggtggacg tcgccactcc cctttcttgc cttcggagtg aggaagcgga   3600 

ggcagcagta cggcagcccg cccagggcca cagagctggg gtcacagcga aacactccga   3660 

aactttcttt tcaattatag ggttcagcct tttttcccat cataacttta attctgtgta   3720 

gatacttcta ttttttattt ttattttttt ttttgagatt gagtctctgt gtcgcccagg   3780 

ctggagtgca gtggcacgat ctccgctcac tgcaggctcc gcctcccggg ttcaggccat   3840 

tctcctgcct cagcctcccg agtagctggg actacaggcg cccgccacca cgcccggctc   3900 

attttttgta tcttagtaaa gacggggttt caccgtgtta gccaggatgg tctcgatctc   3960 

ctgacctcgt gatccgcccg tctgggcctc ccaaagtgct gggattacag gcgtgagcca   4020 

ccgtgcccgg ccttattatt attatttttt tgagacgcag ttttgctctg tcgcccaggc   4080 

tggagtgcag tgatgtgatc tccgctcact gccagctccg cctcccaagt tcatgccatt   4140 

ctcctgcctc agcctctcga gtagctggga ctacaggcgc ccaccaccac gcccggctaa   4200 

ttttttatat tttagtaaag acggggtttc accgtgttag ccaggatggt ctcgatctcc   4260 

tgacctcgcg atctgcccgc ctcggcctcc catagtgctg ggattgcagg cgtgagccac   4320 

cgcacctggc taatttttgt atttttagta gagatggggt ttcaccatgt tgcccaggat   4380 

gttctcgacc tcttgacctc atgatccgcc cgcctcggct tcccaaagtg ctgggattac   4440 

aggcgtgagc caccgcgccc ggccagcacc atcttttcct ttccactgga actgatctta   4500 

ttatttttgc ctccattaga tcatttttgt aacatgtctt gcaggattta ctgtcttgat   4560 

cgtttctctt aacatatttt tttcctgtga tctaaaaaga taaaaaacta tcaattcttt   4620 

tatcaaaagt ggatctagag gctgggcatg gtggctcacg ccagtaatcc cagcactttg   4680 

ggaggccaag gtgggcagat cacctgaggt caagagctcc agaccagcct ggccaacatg   4740 

gtgaagcccc atctttacta aaaatacaaa aattagccag gcgtggtggc acgtgcctgt   4800 

aaccccagct acttgggagg ctgaggcagg agaatccatt gaacctggga ggcagaggtt   4860 

gcagtgagct gagatggcac cattgtactc cagcctgggc aacagaatga gactctgtct   4920 

ccaaaaacaa agtggatcta gaagatcaaa aaagggcatg attccatatt ggcacagcac   4980 

aagccctatt cttggaatta aatggcatcc atcttccgag cccactcctg tcctgcaggg   5040 

ccggcccagc ctgtccctga ggcactggtc cagacaggag cctgtccaca cagctgtcca   5100 

ctcagtgggc ccagtgcttg gcttcacggt cacttgcggc acctagacct cctctggcag   5160 

gtgccattct ttcctctccc tccctgccgc ctcgagtctt tattttctgt gggatcttga   5220 

gtttgataac ctgacctgct gtggtggcag caccgctctg tgtccagatt ctggatgcca   5280 

atttaccaag cgcaggtcaa aaagaagtcc ttgggcagcg gctgcctgcg ttagcttctt   5340 

ggggctgctg taggcggttc caagcaggag agtggcttta aacaacagat gcggatcccc   5400 

tcccggttct agaggcccaa aggctggaat cccatgttgc ccggctgctt ccttctgggg   5460 

cgctctcctg gctcctgtgg ctgcctctgt cttcacatgg cgtcctctct gtgtgtctct   5520 

gcttaaatct ccctctcctt tctcttacaa agacaccagt cattggattt agggcccacc   5580 

ctaatccaat atgacctcat cttaacttga ttacatctgt aaaaacctta ttttcaaata   5640 

aggtcacatt gacaggtact tggggttagg acttgcgctt ttctttttgg gtgacacagc   5700 

ttagcccagc actaactgtg tcaccaggac tgtcgcttga ggcaggaatg aagcacatcc   5760 

tgtttgtaag ctgtcttgtg ccatgcggct gctccgtaca agaattgtta ggaattgatg   5820 

cagtggaatt ttgcatacag tttttcctct cttcagaaac aactttggag aagtaaaggc   5880 

tgaatagcaa tacacaagca ccttatttta ttttatttta gattcagggg cacgtgtaca   5940 

tgtttgtcac atgggaatat tgtgcactgg tggggactgg gcttccggta tcgcatggag   6000 

agggactctt tctgcgctcc cccgcccccg cctccctact gtaaagtgcc cggtgcctgc   6060 

tctctccatc ttcgtgtcca tgggcaccca ttgtttagct cccacttata agtgagaaca   6120 

gtcagtattt gattttctgt ttctgagtta gttcacttag ggtaatggcc tctagctcca   6180 

tccgtgttgc tgcagaggac atgattttat tcttttttat ggctgcagca atacacaagc   6240 

tccttatttt tatttattta tttatttatt tttgttgttt gtttgtttgt tttgagacgg   6300 

agtctggctc tcgtccccca ggctggagtg caatggcgcg atctcggctc attgcaacct   6360 

ccacctcccg ggttcaagcg attctcctgc ctcagcctcc caagtagctg ggactacaga   6420 

cgcccgccac caggcccggc taatttttgt atttttagta gagacaaggt ttcatcatgt   6480 

tggccaggct ggtctcaaac tcctgacttc gtgatccgcc cgcctcggcc tcccaaagtg   6540 

ctgggattac aggcgtgagc caccgcgccc ggccaagctc cttattttaa gcattttttt   6600 

tttctttttt gagacagggt ttcactttgt cacccaggtt ggagtgcagt ggtgtgatca   6660 

tggctcattg cagcctcaaa cttctgggct caagtgacct tcccgcctca gtctcatgag   6720 

tagctgggac tgcaggtgca tgccaccttg gctaattttt attttttgta gagatgggga   6780 

tcttgttgcc aggctggtct caaattcctg ggctcaaacg atcctcctgc ctctgcctcc   6840 

cagagtgccg ggattacagg catcacctag caaagcatta aaacaatttg ctgctgggtg   6900 

cagtaggtca cacctgtaat cccagcactt tgagaggcca aggagttggg gggagttggg   6960 

gggcgggcgg atcacgaggt caggagttcg agaccagcct gaccaacatg gtgaaacctc   7020 

gtctctacta aaaatacaaa aattagccgg gcgtggtgat gcacacctgt aatcccagct   7080 

actcaggaag ctgaggcggg agaatcattt gaacccagga agcggaggtt gtagtgagcc   7140 

gagatcacac cactgcactc cagcctgggt gacagagcga gactccatct caaaacaaaa   7200 

acaaaaacaa aaaaacaatt tgccctgtaa gaactgtcct ctaaaagttt ttggtttttc   7260 

taatgaaaaa tattatggac ttagagaata gaaataaatt tctgcctaca cttccatctt   7320 

ccctcccacc cttctctggc agcccaggag gtctttttgt gtgaatctgc gcagatctca   7380 

gcgtccctgc ccttctttgt gttttgttct ctcttccacc ttaggtcttt ctctggtctg   7440 

ggcacaccca gctgcagggc tcacctttgc ctgtaagaat acagccccca aacacagtca   7500 

gtaccccaag aacagtccct gccatctctg gcggcacaga tgctggccaa gctgcagctg   7560 

ccagtgctgc ccagggagct ggagagctgc cggccaagag cccagcccct ctgggtagag   7620 

caggagccag tgccaccact ccctgtggga ttcggattaa ggacacaccc acccaaagta   7680 

aaccaagctt ggccaaaggc aggtgcccag ctgtggtcac cactccgcag tagttactga   7740 

aaatcttcca tctgcccaat accctcctga gcccgtgaag gagatgagcg gaaagaggct   7800 

ccgcctgttg gaagcacagc caggaaaggt gggctcagat tgctgaagcc tgcaggggaa   7860 

cttgaagaaa gcgtgccagc acaggatggc ggatgatgcc cgcatgacac tcgctcgcct   7920 

ccccggaaca gcctgtggcc ttctcaccta gtgggaagct ccccagccgc gtgtttcagg   7980 

aggtccagca gattcctctg cagaggaatc cctttctgca gagtcggggc tcgctccctg   8040 

ccatctacgg gcagtgctgc ttaaagctgt ggctgcagac cttgcctctg cctgttgaga   8100 

cctcctgcag ggccctccag cccacagggt ccctcagctc tctgggacct gtgaggctct   8160 

ttgggccagc tgcaactgga gctctttgca ggaggggcct ctggcctggc tgaagtcccg   8220 

gcttcctgac tcccctttcc cctcaggtca acaacggttc cagcctcagg gatgagtgca   8280 

tcacaaacct actggtgttt ggcttcctcc aaagctgttc tgacaacagc ttccgcagag   8340 

agctggacgc actgggccac gagctgccag tgctggctcc ccagtgggag ggctacgatg   8400 

agctgcagac tgatggcaac cgcagcagcc actcccgctt gggaagaata gaggcaggta   8460 

ggcggccggc cccacctcct tccccaaagc tgggcttctc tgtcgccagt aacattcagg   8520 

gagcctcagg gctggaaggg acccccggga tcactctgcc tctgcagttt cagctgccac   8580 

gtacgctggt atcacttaat cacttgactg gtctctactt gattccctcc agtgctgctg   8640 

aactcactgc ctaccatttt tgggtgactc tgttagaaag ttcttccttt ctgttgagac   8700 

agaatctcat gtactggtct tgagtccctt gtctggacca acatagaatg gtgtttttat   8760 

ccaattttcc aaatgtgatt ctgatacaaa gattgcagac cacttgtctg gattatataa   8820 

cccaaggggt tctcacactt ggccttgtat catttcaagg acctggagct ttaaatgctg   8880 

gtgcctgcat ctcacctcca gagattctga ttggttggtc tgggcattgc tgggtctggg   8940 

caaagccccc aggtggcact accggtgcgg cccctgcctc cccaagcagg cctggctgac   9000 

tgtcccattg attgaggccc actggtttca cagtgacttt tgcactgtct atacctgaca   9060 

tatttccttt catacattat gctccgtgat tacctataca agaacacaga agtatttgga   9120 

acctcatttc caggtgagga aacccaggtc cagcaaaggg taaatgacta gctccagatc   9180 

acacagcttg tggccatgtt accactggga catggggcca ggccccttct tgaggtgggc   9240 

ctcagccgcc ctcccactgt agggcactga ctccaggtca ccatggtttc cagactgttc   9300 

acctttcctg ttgctgatcc ctgcactctc ctccagcctc cagctccact cccctttgcc   9360 

aaggggctgc ttctatggac aggggctgtc ccgagtggag gctgggggcg agtggaggct   9420 

cacccacttc cagatccagc cctgcgacgc tggctttcag tagtgtgcac attggaatta   9480 

cacgagaaac cttttccaaa tgcaggcctt gggccctact ccagctgcct gcatcaggct   9540 

gttttagggc gggagactgc ccagaggatt ctgacgcagg tagaatccct gccctgaaag   9600 

cctgcaggga tccccggacc ctggtccagg ccttccaagc tcaagggttg cactgccctc   9660 

tggtggctgt gggggagacc aacagctgac ccagccttct gcctcccgcc tgtcttagat   9720 

caggtgcttg aggacgtggc tggagttccc cactagaccg gggtgggggt gggggtgggg   9780 

ggtgggggga ggtgtctgag aatgtctctg ccttctaatc cagccagcat atcttctggc   9840 

tcgccctgaa ctgaggagaa accccagatc cctttgggaa ggtccaggaa gggcaggagt   9900 

ggacaggcac agctctgctg tcagcactgc tgtgggggtg actgtagccc cagtctgccc   9960 

tggtgttttt ctctcgctct tctccatgcc ggcctttgcc tctagactga gaaaccgggg  10020 

ttgactcaag tggcacctgc aaaagtgatc atggcagttc acttagcctg caggtgacag  10080 

ggactgtgaa tctagtccct ggcgagcctg gaaagagggg caaggtagag gctctggctg  10140 

ccggggtttc tttggtgagt ccgttcactc ggctggacac agacggatca ggaaagattc  10200 

ctgttgctac tcggctggtg gccagaggga gagaggacgt gtccgtaact gaagcaaggt  10260 

ggataagctt cgggaacgag cgaggcacag attcggtgct gggggagtga tgaggtgctg  10320 

gaggagctgg gtgctctgct ctgcagggaa tcaggaaaac tttggggctg cagctccaat  10380 

tgagctgggc cttgggggtt gggtatgttt ggttccttgg aaactgggaa gagggaatgg  10440 

ccatctttta agcaaaagcc cagcggctat aaatgctaca gtgaggctgg gtgcagtggt  10500 

tcacgcctgt aatcccagca ctttgggagg ccaaggcagg tggatcatga ggtcaggagt  10560 

tcaagaccac cctagccaag atggtgaaac cccgtctcta ctaaaaaaaa tatataaaaa  10620 

ttagccaggc ggggtggcgg gtgcctgtaa tcccagctac ttgggaggct gaggtagaga  10680 

attgtttgaa cccgggaagc ggaggttgca gtgagctgag attgtaccac tgcactccag  10740 

cttggggaac agagtgagac tatgtcttga aaaaaaaaag aaaaaaaaaa gctacagtga  10800 

gtagttgagt ttgcctagga agcgtggaag ttaagtcaga cgtactttca ggctgggtca  10860 

tgacttgtca cttaagcaga gatgagcact tgagaggttt tgaagagaag tgatgtggca  10920 

gccttactgc atgttccatg gacagactcc agggaggccg tgaaaccccc agagcacagc  10980 

ttctaagaac gtgcccactc cttagcacgt cacttctccc aaccctgccc tgctctgagg  11040 

tctgtgctgt gaaggtggcc gagtagactg gacggcaggg agtggggctg tcatcatcag  11100 

atgagagcta aggggacccc caccagggtg gcggcaatgg cagagggtag gcaaaacgct  11160 

tgtatttgca acataaggtg agatttgaca gctgaccgag ggtgggagca gcagccaaaa  11220 

ccaaaaaagc cagagggaag ttgcaagcac agaaaaaata gaagatttaa tgggagaaat  11280 

aacaatagct ggcatctatt gaacacttac tgggagctag gtacagggcc cattcattca  11340 

ttcatgcaat taaaactttt tttaagaaac ggggtcttgc tctgttgccc aggctggagt  11400 

gtagtggtat gatcacagct cactgcagcc ttgaattcct ggcctcaagg agtcctccca  11460 

cctcagcctc ctgtgtagct gggattatag gtacgtgcgg tacacctggc tccctttaaa  11520 

agttttttgt agaggcaggg cacagtggct cacacctgta atcccagcac tttgggaggc  11580 

caaggcagga ggatcacaag gtcaggagtt cgagaccagc ctgaccaaca tggtgaaacc  11640 

cgtctctact taaaatacaa aaattagccg ggtgtggtgg cgggcgcctg taatcccagc  11700 

tactcaggag gctgaagcat gagacttgct tgaacccagg aggcgaaggt tgcagtgagc  11760 

cgagatcgcg ccactgcact ccagcctggg tgacagagca agactccgtc tcaaaaaaaa  11820 

aaaaaaagtt tcttgtagag gcagggcctt gctttgttgc tggtgcaatc acggctcact  11880 

gcatcctcta actcctggcc ttaagcaatc ttctgtcctc agcctcccaa agcactggga  11940 

ttacaggcat gcatgaccac acctggtccc tgccattgtt tattgagcac ctactgagtg  12000 

ccatgtatta agtgctgggt atttgtcagt ggacaaaaca gattaaaaaa atcacagccc  12060 

ttagggagct taccttctgg caggggcgtc agacaataac acagcaagtg ctgaggaaga  12120 

aacggaggcg gcagggagcg tggcagttga gcgtggcctt catggagctg cgacagtggt  12180 

actcgggcag gggcagcacg gaggctgtgc gccagaggag gaggactgag gggcaagggg  12240 

gagagctctg gttggaaagg caggggagat tctccagggc cttgccggtg ccagtgacaa  12300 

ctggggtttt cctgagacgg gactgcgagg aatgggggct ctcaggcttg agagggcaaa  12360 

agtgggtctg ggatgccgtc tgcccacaga gccccttccc caacggctgc ccaggccaag  12420 

gccaaccctg ttgggttgtg tggtgtgagc catgaagccg ctgccaggct tgtacctcag  12480 

gcgtggtcgt gatgccccag cttcaccggc cctgcctgtg gggacgtggt gcctgtgtgc  12540 

gggagcctgg gcctcagccg aggccctgag ctccggcact gcccagaacc cagctcagcg  12600 

ctggtactca gcccgcccgc tgtggccctg gtggagtgga gcacgtgccc agtgggggct  12660 

ggccttgtcc catcgcggac ctgtcctttc ccggggcagg gtggtgtggg agagggtatc  12720 

agggacattt tctgagtctg ctctgtctct gccgcccctg cctgaacaca gattctgaaa  12780 

gtcaagaaga catcatccgg aatattgcca ggcacctcgc ccaggtcggg gacagcatgg  12840 

accgtagcat ccctccgggc ctggtgaacg gcctggccct gcagctcagg aacaccagcc  12900 

ggtcggagga ggtgagtgag ggcctgagga ccgcgtgggc gggcaagtga gccaaggggg  12960 

cctgtcccct gcctctcacc aggcagccca ctgtcccgtg aggccactca actcgtgact  13020 

gtcaggtcca gaactctgac gaagtaactg gacgtagggt atggttcatt gccttgcaga  13080 

agatttcagc tggttgacat cgaggaaacc tgaaccttaa atcagagtaa agagtttagg  13140 

ggtaaaagcc tctaaaagat gaacgaagca tgtttggcca acagaagaaa cagacgcttc  13200 

ctttggttgt agggagttta ataatggtgc cagtgagaac cgtaagccct gggagtggtg  13260 

cctgctgctc tgctgagctc cttggttgga atccacacaa ctttctgagc tctaccatct  13320 

gcttggcact gttggggata caagattggt ccggggcact gtgtccccag aacacttagc  13380 

ggaaagaact acatcctccc aactgccaaa tgcaggcctg tagcggtagg agctgagagg  13440 

agagaaagtt ccactttttc gactctacca gctgaaaatg caggcgtcct cacctcctag  13500 

aaatccaatc atgcttctgt tcagtggggc cagcctgtga tgtcccagca gctgcctaga  13560 

acgcaggagt ggctggcgca ctcccatgta actctgcatg tgcgccgacc gcctgacggt  13620 

ccttgccagc cttgtagtct gtctagtgtc ccccaggaac ccccttcctc ctgtccattc  13680 

agctaggtct gcaccaataa aatgggccta aggcgtcgca ggtggtcact agttctggac  13740 

tcgaagtgcc ttgggcgcag ggatgaccca ggcttcttgt atcccatcac cgtctaacag  13800 

tgggcacatg ggctcaccac acatgcgttt gcttaccgag ccccctgcag ggagtgattg  13860 

cagtcttccc tttccattgc ctctcagaac tcaactgttt ctcattcttt ccgcccagca  13920 

gccctggata cttaataagt actttgaagt gcttcttcat actggggact gtctttcctt  13980 

tgagagggaa gagtattagt aaaccaggtt ctgtgtgccc ctctgtgcag gac cgg     14036 
                                                        Asp Arg 
                                                         1 

aac agg gac ctg gcc act gcc ctg gag cag ctg ctg cag gcc tac cct    14084 
Asn Arg Asp Leu Ala Thr Ala Leu Glu Gln Leu Leu Gln Ala Tyr Pro 
         5                   10                  15 

aga gac atg gag aag gag aag acc atg ctg gtg ctg gcc ctg ctg ctg    14132 
Arg Asp Met Glu Lys Glu Lys Thr Met Leu Val Leu Ala Leu Leu Leu 
     20                  25                  30 

gcc aag aag gtg gcc agt cac acg ccg tcc ttg ctc cgt gat gtc ttt    14180 
Ala Lys Lys Val Ala Ser His Thr Pro Ser Leu Leu Arg Asp Val Phe 
 35                  40                  45                  50 

cac aca aca gtg aat ttt att aac cag aac cta cgc acc tac gtg agg    14228 
His Thr Thr Val Asn Phe Ile Asn Gln Asn Leu Arg Thr Tyr Val Arg 
                 55                  60                  65 

agc tta gcc aga aat gtaagaaccc ttgaggtcag ctccttccct gcctgccgcc    14283 
Ser Leu Ala Arg Asn 
             70 

catgcccttt tctctggaag gttgagaagc ccagcggggc ccctgcctct gatgccagca  14343 

caagggttac aggctgtcct gctcgggttt ggttttgctg ttgtgagcta gaaagctgtg  14403 

tgtaaaggtg acgaagagca cccagagtcc tttggagctt tagcagctta ctattggaga  14463 

catgctccat tcagaggggt ggcaaaggct cacgtcacac tcctggtggg gtcctcaagg  14523 

cacaagcagg tacagagtgg aaggaagggg ctggagggct cacaatgagc ttttcagacc  14583 

tctcaccttg ccataaaaaa taagtgtaat gtggccagtg cggtggctca tgcctgtgat  14643 

cccactgctc tgggaggcca aggcaggtgg atcacctgag gtcaggagtt ccagaccacc  14703 

ctggccaaca gggtgaaagc ccgtctctac taaaatacaa aaattagccg ggcatggtgg  14763 

cgcacacctg tagtcccagc tactcaggag gctgaggcag gagaactgct tgaaccctgg  14823 

aggcagaggt tgcagtgaac tgagatcgca ccactgcact ttagcctggg cgacagagca  14883 

agactccatc tcaaaaaaaa ggtgtaatgt gaaccaaaac gagtagtcaa aaaagggggg  14943 

gaactgtctg aaatcttttc cagagcacat ctgtcccata accaggtatt acaagtcaca  15003 

gtctaaaggc tgggcatggt ggctcaagcc tgtaatccca gcgatttggg aagcagaagc  15063 

agtgggattg cttgaggcca ggagtttgag acaaaactga gcaacatggc gagaccctgt  15123 

ctctaaaaaa tttataaaaa taattagctg agggccaggc gcggtggctc acgcctgtaa  15183 

tcccagcact ttgggaggcc aaggcaggcg gatcatgaag tcaggagttc aagaccagcc  15243 

tggccaagat ggtgaaaccc cgtttctact aaaaatacaa aaaaaattag ctgggtgtgg  15303 

tggcgggcgc ctgtaatccc agctactcag gaggctaagg caggagaatc gcttgaaccc  15363 

tggtggcaga ggttgcagtg agccgcaatc acgccactgc actccagcct ggatgatggg  15423 

gtaagactgt ctcaaaaaaa aaaaaaatta gctgagcatg gtggcgtacg cctgtagttc  15483 

acgccgtcat ggaggttgag gcagctcctc aggaggctgg ggcagaagga tctctttgct  15543 

tgagcccagg agttcaaggc tgcagtgagc tgattgtgcc actgcactcc agcctgaaca  15603 

aaaacaagac ctgtctctaa aaacaaacat acagtgttca caatgctgcc caagaagggc  15663 

cagtttttgc agctgccccc atgtagcaaa atctggtgct tctgtttcat agacccaaat  15723 

ggaaattaag tggatgtgtc ttatttgtaa atttaaaaat attagcgaat gtttgggaat  15783 

tttttttttt tttttttttg agacagaatt ttgctcttgt tgcccaggct ggagtgcaat  15843 

ggcacgatct cagctcacca caacctctgc ctcccaggtt caagcgattc tcctgcctca  15903 

gccccccaag aagctgggat tacaggcaca caccaccatg accggctaat tttgtatttt  15963 

tagtagagat gaggtttctc ccatgttagg ctggtctcga actcccaacc tcaggtgatc  16023 

cgcccacctc ggcctcccaa agtgctggga ttacaggcgt gagccactgc gcccggccta  16083 

atgtttggga ttttatgaca tgtcagaagc attacttcag gctttggttt ttaagtaaaa  16143 

tagcatctaa tcctctactg agaactcata agaaaacatt ccttatatgc tgtggtcttc  16203 

agttatacaa gcattttaaa aacaggagaa tgaatataaa tcttaaatca ggcattaaac  16263 

ccagctgaat tgttggaagg aggtaagcct gagaccattc ctggacagct tttaccaaca  16323 

cccatgtaaa gggggaaagg gtgggcaaga cgtgtgcagc agtctgtatg gacagcttac  16383 

cagagactga gggctgaggc agaatcgtga ttcctctgac ccagcagggg cctcctgaca  16443 

ccgtcagtgc cttggagatg tgaataccca cctcaccgcc tgaacggcct gtttttgcag  16503 

ttgcccccat gtagcaaaaa gtaggatgca cggataggac ttcaggggtc tggagaacat  16563 

gtttttgcat aaaccccagc tttgctctac tgtggcacag agctctggag cctggtttgt  16623 

gaatgagcct agctgattct ggctttttct cctttcttgc tctaggggat ggactgaacg  16683 

gacagttcca gaagtgtgac tggctaaagc tcgatgtggt cacagctgta tagctgcttc  16743 

cagtgtagac ggagccctgg catgtcaaca gcgttcctag agaagacagg ctggaagata  16803 

gctgtgactt ctattttaaa gacaatgtta aacttataac ccactttaaa atatctacat  16863 

taatatactt gaatgaaaat gtccatttac acgtatttga atggccttca tatcatccac  16923 

acatgaatct gcacatctgt aaatctacac acggtgcctt tatttccact gtgcaggttc  16983 

ccacttaaaa attaaattgg aaagcaggtt tcaaggaagt agaaacaaaa tacaattttt  17043 

ttggtaaaaa aaaattactg tttattaaag tacaaccata gaggatggtc ttacagcagg  17103 

cagtatcctg tttgaggaaa gcaagaatca gagaaggaac atacccctta caaatgaaaa  17163 

attccactca aaatagggac tatctatctt aatactaagg aaccaacaat cttcctgttt  17223 

aaaaaaccac atggcacaga gattctgaac taaagtgctg cactcaaatg atgggaagtc  17283 

cggccccagt acacaggggc ttgacttttt caacttcgtt tcctttgttg gagtcaaaaa  17343 

gaaccacttg tggttctaaa aggtgtgaag gtgatttaag ggcccaggtc agccactgtt  17403 

tgtttacaaa atcaggtaac taactgcata cactttttct ctttccatga catcaagact  17463 

ttgctaaaga catgaagcca cgggtgccag aagctactgc gatgccccgg gagttagccc  17523 

cctggtaata gctgtaaact tccaatttct agccatacgc tcagctcatc catgcctcag  17583 

aagtgcatct ggagagaaca ggtttctaag cataaaagat gaaagagcag ttggactttt  17643 

taaaaattca gcaaagtggt tccctctctt agggacagtc aaaaccaagt cacttaggta  17703 

gtaccaaaat aaataaggaa aagcttagct ttagaaacag tgcaacactg gtctgctgtt  17763 

ccagtggtaa gctatgtccc aggaatcagt ttaaaagcac gacagtggat gctgggtcca  17823 

tatcacacac attgctgtga acaggaaact cctgtgacca caacatgagg ccactggaga  17883 

cgcatatgag taagggcact gacggactca tgatttcttc ttaccagatg ctttcctgtt  17943 

ctttaagagt ttaaaatcat cagaaaggaa aaacaaactc tatattgttc agcatgc     18000 

 
           
             666  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            666 

ctttcagaat ctgcctctat                                                 20 

 
           
             667  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            667 

agtccatccc atttctggct                                                 20 

 
           
             668  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            668 

actgtggtga gtctcccacc                                                 20 

 
           
             669  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            669 

agtgtcccag tggcgacctg                                                 20 

 
           
             670  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            670 

cacagtccat ggcctgggca                                                 20 

 
           
             671  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            671 

ctccgcttcc tcactccgaa                                                 20 

 
           
             672  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            672 

tactcgggag gctgaggcag                                                 20 

 
           
             673  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            673 

ccgtctttac taagatacaa                                                 20 

 
           
             674  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            674 

tcaagacagt aaatcctgca                                                 20 

 
           
             675  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            675 

ctttttagat cacaggaaaa                                                 20 

 
           
             676  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            676 

gccatttaat tccaagaata                                                 20 

 
           
             677  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            677 

ggcccactga gtggacagct                                                 20 

 
           
             678  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            678 

gcatctgttg tttaaagcca                                                 20 

 
           
             679  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            679 

acggagcagc cgcatggcac                                                 20 

 
           
             680  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            680 

ggtttcacca tgttggtcag                                                 20 

 
           
             681  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            681 

tctcggctca ctacaacctc                                                 20 

 
           
             682  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            682 

agggacgctg agatctgcgc                                                 20 

 
           
             683  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            683 

ggtctcaaca ggcagaggca                                                 20 

 
           
             684  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            684 

atccctgagg ctggaaccgt                                                 20 

 
           
             685  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            685 

caaacaccag taggtttgtg                                                 20 

 
           
             686  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            686 

gaagccaaac accagtaggt                                                 20 

 
           
             687  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            687 

tgcggaagct gttgtcagaa                                                 20 

 
           
             688  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            688 

gggagccagc actggcagct                                                 20 

 
           
             689  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            689 

cgggagtggc tgctgcggtt                                                 20 

 
           
             690  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            690 

gctggacctg ggtttcctca                                                 20 

 
           
             691  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            691 

aagcagcccc ttggcaaagg                                                 20 

 
           
             692  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            692 

agggctggat ctggaagtgg                                                 20 

 
           
             693  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            693 

agaaggcaga gacattctca                                                 20 

 
           
             694  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            694 

gcccttcctg gaccttccca                                                 20 

 
           
             695  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            695 

ctcagtctag aggcaaaggc                                                 20 

 
           
             696  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            696 

ctgatccgtc tgtgtccagc                                                 20 

 
           
             697  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            697 

aagtagctgg gattacaggc                                                 20 

 
           
             698  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            698 

ggccctgtac ctagctccca                                                 20 

 
           
             699  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            699 

atcataccac tacactccag                                                 20 

 
           
             700  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            700 

ttgtatttta agtagagacg                                                 20 

 
           
             701  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            701 

acaaggccag cccccactgg                                                 20 

 
           
             702  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            702 

ggcagagaca gagcagactc                                                 20 

 
           
             703  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            703 

tgcctggcaa tattccggat                                                 20 

 
           
             704  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            704 

cccgacctgg gcgaggtgcc                                                 20 

 
           
             705  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            705 

gatgctacgg tccatgctgt                                                 20 

 
           
             706  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            706 

acctcctccg accggctggt                                                 20 

 
           
             707  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            707 

ccagggcagt ggccaggtcc                                                 20 

 
           
             708  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            708 

ctagggtagg cctgcagcag                                                 20 

 
           
             709  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            709 

tgtctctagg gtaggcctgc                                                 20 

 
           
             710  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            710 

cggagcaagg acggcgtgtg                                                 20 

 
           
             711  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            711 

aaattcactg ttgtgtgaaa                                                 20 

 
           
             712  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            712 

tgcgtaggtt ctggttaata                                                 20 

 
           
             713  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            713 

agagcagtgg gatcacaggc                                                 20 

 
           
             714  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            714 

tgttggccag ggtggtctgg                                                 20 

 
           
             715  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            715 

agctgtccat acagactgct                                                 20 

 
           
             716  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            716 

cttctggaac tgtccgttca                                                 20 

 
           
             717  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            717 

gttgacatgc cagggctccg                                                 20 

 
           
             718  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            718 

atagaagtca cagctatctt                                                 20 

 
           
             719  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            719 

tgtagattta cagatgtgca                                                 20 

 
           
             720  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            720 

ttaagataga tagtccctat                                                 20 

 
           
             721  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            721 

tccttagtat taagatagat                                                 20 

 
           
             722  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            722 

tagttcagaa tctctgtgcc                                                 20 

 
           
             723  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            723 

ccggacttcc catcatttga                                                 20 

 
           
             724  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            724 

aaaagtcaag cccctgtgta                                                 20 

 
           
             725  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            725 

aagttgaaaa agtcaagccc                                                 20 

 
           
             726  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            726 

gtaaacaaac agtggctgac                                                 20 

 
           
             727  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            727 

gtatgcagtt agttacctga                                                 20 

 
           
             728  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            728 

tgatgtcatg gaaagagaaa                                                 20 

 
           
             729  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            729 

tttagcaaag tcttgatgtc                                                 20 

 
           
             730  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            730 

tgtctttagc aaagtcttga                                                 20 

 
           
             731  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            731 

aacctgttct ctccagatgc                                                 20 

 
           
             732  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            732 

tagaaacctg ttctctccag                                                 20 

 
           
             733  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            733 

tgcttagaaa cctgttctct                                                 20 

 
           
             734  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            734 

aatttttaaa aagtccaact                                                 20 

 
           
             735  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            735 

tgttgcactg tttctaaagc                                                 20 

 
           
             736  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            736 

agcttaccac tggaacagca                                                 20 

 
           
             737  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            737 

gggacatagc ttaccactgg                                                 20 

 
           
             738  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            738 

tttaaactga ttcctgggac                                                 20 

 
           
             739  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            739 

gacccagcat ccactgtcgt                                                 20 

 
           
             740  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            740 

gaagaaatca tgagtccgtc                                                 20 

 
           
             741  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            741 

gattttaaac tcttaaagaa                                                 20 

 
           
             742  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            742 

tagagtttgt ttttcctttc                                                 20 

 
           
             743  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            743 

aatatagagt ttgtttttcc                                                 20 

 
           
             744  
             791  
             DNA  
             Mus musculus  
             
               CDS  
               (19)...(606)  
             
           
            744 

agctacacag cttgtgcc atg gac tct gag gtc agc aac ggt tcc ggc ctg       51 
                    Met Asp Ser Glu Val Ser Asn Gly Ser Gly Leu 
                     1               5                   10 

ggg gcc aag cac atc aca gac ctg ctg gtg ttc ggc ttt ctc caa agc       99 
Gly Ala Lys His Ile Thr Asp Leu Leu Val Phe Gly Phe Leu Gln Ser 
             15                  20                  25 

tct ggc tgt act cgc caa gag ctg gag gtg ctg ggt cgg gaa ctg cct      147 
Ser Gly Cys Thr Arg Gln Glu Leu Glu Val Leu Gly Arg Glu Leu Pro 
         30                  35                  40 

gtg caa gct tac tgg gag gca gac ctc gaa gac gag ctg cag aca gac      195 
Val Gln Ala Tyr Trp Glu Ala Asp Leu Glu Asp Glu Leu Gln Thr Asp 
     45                  50                  55 

ggc agc cag gcc agc cgc tcc ttc aac caa gga aga ata gag cca gat      243 
Gly Ser Gln Ala Ser Arg Ser Phe Asn Gln Gly Arg Ile Glu Pro Asp 
 60                  65                  70                  75 

tct gaa agt cag gaa gaa atc atc cac aac att gcc aga cat ctc gcc      291 
Ser Glu Ser Gln Glu Glu Ile Ile His Asn Ile Ala Arg His Leu Ala 
                 80                  85                  90 

caa ata ggc gat gag atg gac cac aac atc cag ccc aca ctg gtg aga      339 
Gln Ile Gly Asp Glu Met Asp His Asn Ile Gln Pro Thr Leu Val Arg 
             95                 100                 105 

cag cta gcc gca cag ttc atg aat ggc agc ctg tcg gag gaa gac aaa      387 
Gln Leu Ala Ala Gln Phe Met Asn Gly Ser Leu Ser Glu Glu Asp Lys 
        110                 115                 120 

agg aac tgc ctg gcc aaa gcc ctt gat gag gtg aag aca gcc ttc ccc      435 
Arg Asn Cys Leu Ala Lys Ala Leu Asp Glu Val Lys Thr Ala Phe Pro 
    125                 130                 135 

aga gac atg gag aac gac aag gcc atg ctg ata atg aca atg ctg ttg      483 
Arg Asp Met Glu Asn Asp Lys Ala Met Leu Ile Met Thr Met Leu Leu 
140                 145                 150                 155 

gcc aaa aaa gtg gcc agt cac gca cca tct ttg ctc cgt gat gtc ttc      531 
Ala Lys Lys Val Ala Ser His Ala Pro Ser Leu Leu Arg Asp Val Phe 
                160                 165                 170 

cac acg act gtc aac ttt att aac cag aac cta ttc tcc tat gtg agg      579 
His Thr Thr Val Asn Phe Ile Asn Gln Asn Leu Phe Ser Tyr Val Arg 
            175                 180                 185 

aac ttg gtt aga aac gag atg gac tga ggagcccgca caagcccgat            626 
Asn Leu Val Arg Asn Glu Met Asp  * 
        190                 195 

ggtgacactg cctccagagg aaccgcgacc atggaaagac cttggcctga agacaggtcc    686 

cagagaacag ctgtctccct atttccaggt ggtgggaacc ccaagctggt gattcactgg    746 

acatctctgc gttcagcttg agtgtatctg aagagtttac gccgg                    791 

 
           
             745  
             30310  
             DNA  
             Mus musculus  
             
               CDS  
               (19791)...(19802)  
             
           
            745 

gctcgctttg ggtcatgatg tttcattata ggaatagtaa gccaaactaa gatgatgtct     60 

cttcacaaca ttagaaaagt gactaagact ggcctctata gactcatacg tttgaataga    120 

actatttggg aaggactagg agatatagcc ttgttggaga aggcgtgtca ctgagggtgg    180 

gctttgaggt ttcaaaagcc cagagtcttt ccttctctat ttcctaactg cagataggga    240 

tgcaagctct cagtgattcg ccaccaccat gtctgcctgc ctcttgccac gttccctgcc    300 

atgatggtca tggactctaa ctctatgaaa ccataagccc caaattaaaa gaaaaaaatt    360 

gagagagagt ttttttctgt atagacctga ctgttccaga atcactcggt acgacacgac    420 

gcgaagctgg ccttgaactc agggatcctc ctgcctctgc cttccaagtg ctgggattaa    480 

agggatgtgc caccactact caactaaatg gtttctttta taattcatcg tggtcaaact    540 

gttttgtcat ggtaacagaa aaacaactaa gacccagcca tgtctgaggc acacacattt    600 

atagatgtac agttaagctt tttctaattc tgtaatggag acagactcac acaatagtac    660 

cgcctggaat gttggggatg ggttctaatg cattatctta attcagctca caaagtcaca    720 

tgggaatcta catgttcaca tgctgagggt ccctgtcccc agttggtttt tgattgatca    780 

ataaagagcc aatggctagt ggttgggcag ggagaaagag gcaggacttt taggatttcc    840 

aggcaagaaa ctcaggggag aagatgaaag gactctacca tgagaggggt gtaggacgga    900 

ccacaccatt gacagggaag cagaaagatc agacttaaag gcctgccaac atgtaagaat    960 

ccagaaaggt gactccaggg gccattgatt gggtctgggg tcacagagat aaaataaaga   1020 

tttgtcaagt attaactcaa gaataccaga ggggagtgtg tgctagccta ggggagtttt   1080 

ggaaataccc aacgtttgaa ctagtcaaga catctcaaaa tataaaggtt gcatgtatgt   1140 

gtctttcatt cgcaaatcca gagagctctg gcgggtggct agaagtgtga tcactttctg   1200 

ggaactcaga gtggattaac aattcaccat tacaagtgca gtttttggta gggaaggtca   1260 

tgtttgtaat ggtgccgagt caccaaagaa agagaaacag ctcttagagt tctatgccag   1320 

agggcagagg agcatgcaac ccatccttca gggtttgaca agcagaaggc aggctggtgg   1380 

cacagaaaaa aatcatagtt ctggactagt ctgggctaca tagtaacctc tgtcttaacc   1440 

ttctcccctt gccctaaagc atctatgatc tgtattggtg ggagcgagag ctgggtggtg   1500 

ctgaggttag aaggctccct agctatgggt atttgttaaa atgtgaactc ctccaagaga   1560 

tgttataaag tggaaatgtc tagtctcttt ggaaagttag ttatgacaaa tgacattttg   1620 

ctggggcaca caagtgaaag gatgtcttcc taaagcagac acaggaaaga atgttttccg   1680 

gaagcagaca caggtaaaag gatgttttga tatagcaaac atgtaaaagg acccttgaca   1740 

aaggagtata aatatgaccc cacagaccac aggagatgag cactgagcct tggtttggtt   1800 

tgttctgcct cgctgttctt cgctaactac atacatgcat tggtttacct tatatagtgt   1860 

tgttaatcgc aacttgtgga aacaccacca ttgagagaaa gagcagtcca ccaaagaact   1920 

gcttgtgagg ttcctacagc agcttgctgc ttctgcggcc tcgcctcagg ctgcttggtg   1980 

agcctagcag tttcttcgac tggactgtcc ttgccagttt gtgtgtggtg tctgtctgct   2040 

tagaagtctg atctgcagct gctgagttct atttggcgtt tgctacgaga ctgaactgcc   2100 

cccaaagaac tatggcaccg tccacttccc ccatagccta attttctctt ctcccacctc   2160 

tgctgggtgg tgggctagag gagacgttga acctttatta aaagtaggtt gcaaaaaagt   2220 

tgagcctaca aggttatata ttcagaacaa tttctggaat acgattgggt ctacgtggtc   2280 

ctagaaatat tcaggggcaa agaacacgca gcttgtgtgc gccaggttct gctggctggg   2340 

tggagagagc gtgccaggta gcacagtgtg ccaggcagca cagagccttt gccctctccc   2400 

accctagccc atccctattc cttgtgtcac aggaagtatg gagctaggac cagggaggtg   2460 

attgttctgt gatctctaat gtttaggtga gaaatgcccc ttcacaccag acctttgtgt   2520 

tcacaccagg cccctgggtt cacaccagtt acacttattt taatgaagct ctttctgtct   2580 

aaaatttcta gctcctccct ttaacacttc ctaatttaga gattatttag gctgcacatt   2640 

aaaactggaa gtttcactga tagttcagtg gtaaggttgg actcatttaa agtgaaaatt   2700 

ggattcccag caaccacacg gtggcccaca gccatctgta atgggatccg atgccctctt   2760 

ctgatgtggc tgaagacagc tacaatgtac tcatatacat aaatgaataa ttaaagtgaa   2820 

aattggtatg ttccatcttt atgaagttgt gaaatcagtt tccctttttc atttgcattg   2880 

attgccaagc acctcggaga gaatcccagt taaaaatatt acgtgttcag gtcatgatca   2940 

tgcacgcctt taatcccaga ggcaaaagca ggaggagctc tgtgagttct aggccagcct   3000 

ggtttgcata gctagttcca ggccagtcag ggctacatag tgagagcctg tctaaaaaaa   3060 

aaaacaaaac aaaaacaaaa cttttttctc attattttcc actttgaaat ctagataatt   3120 

cagcttgcat gttttaaatt taaaaactct gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt   3180 

tgcctgcata tatgtgcacc acatgtgtgc ctggtcctca tagaggccag aaggggggtc   3240 

agtcccttgg aattagaata acagatgatt gtcagccacc atatgggtgc taagtactga   3300 

acccagatgg atgctctgta agagtgagaa gtgcttttaa ccagtgagcc atctctccaa   3360 

ccctgccccc gctgttcatc accaagctct tccactatgt gatttcaagt gtaacttttt   3420 

ttttggcggg gggtgggggg gtgggggagt ggggggtggg gtggggttgg tttttcgaca   3480 

gacagggttt ctctgtatag ccctggctgt cctggaactc actttgtaga ccaggctggc   3540 

ctcgaacaga ggcctcccaa gtgccgggat taaaggcgtg cgacaccacg cccggcttca   3600 

agtgtaactt ttattgatcg taaaattaga gccatcttcc tttaagaaga attggaaaat   3660 

ataaagagga aaaagaaacc ctggagatgg ctcggtttgt aaagtacttc atatgcgtaa   3720 

gaactggact ttggatccct agcacccatg taaaaactag agtgctgtgt gtatctacaa   3780 

ttccattgtt attggtgcac ggtggaagct tcctggagct cacctggcag tcagcctagg   3840 

gaaatcacgc gtggagctgg gaagctggtc cactcccctc accccacacc atctcaaaag   3900 

aaaaaaaaaa aggtggaaag gtggagagtg atgaaggaaa acactgacct ctggcttaaa   3960 

tacacacata cacacacaaa cacacaccaa ccatgtgatt tttttttttt tttttgtctt   4020 

ctcagatcca gtttctctgc tcaggaacag caatttccat ggttctattt acttcctcat   4080 

acttccagaa ttcactttct tgtttctctt tcacttttgt cactgccacg tgtcctttgg   4140 

gggtactggc tggcacttaa gtatatagca ttgggacttc tctggacagg ggaactagct   4200 

agcagtttga gattatctgc tagcctcctg gttctttcca cattcatcct tgctgattca   4260 

ttccatgacc gagaaccccg caacccccat ccctgccttc cccacaagag tttaaaaatt   4320 

ctgcaagcag ctgcgcagga gaaacaatag ggacctccca gcatctctga tagggccgat   4380 

tctgacaggg tcactagtct tgagtgtgcc aaccctgcta tgtaatacat caagacaatg   4440 

cggagaggtc gggatcaagt atgacacccc atcctcacga gggcaggtcg cccaggcttt   4500 

ggggactctg gggagcgcag gttccgggtg taccttcctt cctgtccccc gtagcgagcg   4560 

ggtaggaccc ttgggtttcc gcaaagtgtg gccagtcgga gggcggagca tccggagggc   4620 

ggggctatca caggggcggg gctcccgggc gagcacgagg aaaggtaggt ggagtagagc   4680 

gccgggccga gtgtggctcc gcaaaccttt gccttagccc gttcgccgcc cggtaccggc   4740 

gcagcggcgt ctgcgtggtg agtatgccca ccctactggg cgcccccacg gttcccctct   4800 

gggaggacgg ggtcggcacg gagctcagtt tcgtatgcta tcgatccttc gtgatggcgg   4860 

ggctcttgcg ccttgatgga ggcggggtgg gggcgccggc cacagggtgc caccgcggag   4920 

ctgaggggaa ggcactcact cgaaggcctg gggcgtgcgc cactcgcggt ccccctcagc   4980 

gctcggtcct ggtccgcttc gggcaggcct cctggtggac ccggggtccc cgcggtcgcg   5040 

cgccactcgg caggtgcgcg cagagctgga aaggcgggcc tgaggtctcg ctgcgctccg   5100 

ctatggccac ccacaaaaat caacaaggaa cggctacagc ccacaaatgg gccctgcaaa   5160 

agccctggaa ccccaaccca gggaacacag accttggaag actgcagcga ggggcacctt   5220 

tcctacaccc gtgggcacta ctgtgtgcac agctcacact cacgcctgaa ctgtgaggaa   5280 

gtggctgacc cctccgcatc tccagtaccc aaaatggttt gaaaatgtgc acagactggt   5340 

tgctgatgtt tttaaaaagt ttgttgaatg gttggctgaa taaccctata ggattctaga   5400 

agaaacccac agccttcagc caccaagtgg cctgggccca caaggattca cacattcgtt   5460 

cattcattct ttcgtacatt catttacata ctcaacaaat aagtgtggac cagggacgga   5520 

tcagggtaga actttgtggg tggtgagagg ctggaatgaa gagctctgta aaggaccagg   5580 

tggtgttgag tatgggactt ctaggctggg cttgaccttc atctgataag ccacatagtt   5640 

ctgagtcaag agcatcctga ggacccaggc agggctcccc tactttccca ggctactgcc   5700 

tggtgccatg gccaggattg cccttactgg aagactacct tgaagccggg tctaggataa   5760 

gctagctgtg gaatggagct gggagaaacc acaagaagga tgtggacttt ccacattcca   5820 

gctctaccca accaggagac tttgcagccc tgccccatcc cctgggactt ggtcccaggc   5880 

actaccctgg cagtcagctc tgagtgtttc catggggggg ggggggggag cctgatccag   5940 

tgctggggct gagttcagag gctttaatac ttgagtgggc tgagctctaa gaaggactcg   6000 

gctgggtggt ggtggggaag cagggtggcg attgtgtgtg tcctggcctc tactgcctct   6060 

cttgcccaga gagggaatgg cagggaggtt ggcttattac agctgggtta gcaggcattt   6120 

cacccactga cgaaaggtgc tatctcctgg ctactgcggg gtggagttgg gtacaggctt   6180 

tggtgatggc aagtgaagag aagccggctg gatgtggcat gctctataaa gagatttaag   6240 

tagccccaag gtggccaggt tactggagct ctgaaggatg agttgagggt gtacctgaaa   6300 

agtgggctgt tagggcagtt actggcgagg ctgggggagg ggaagtgatg ctcacagctt   6360 

gaggttacct ggttcctctt atttgcaaga aagaatagcc tacggggggg gggggggggg   6420 

cacagtgctg ggtgcctggc ctccggaagg aaggcctgat gacacagcct tttagacctt   6480 

ccgaagggca ctgcatgctt ttccagctgc ccttttgctc tctagtggga agctgagggt   6540 

tggggaccca catctaggct gtgttcaaga ccaaagagcc attcctcatc agggagacag   6600 

tgaatctgat ggttccaagg atgagagttg gaaactgccc gtccataaga agcccccact   6660 

gtgggtctgt ggtcactgga cattttgtct gtggttgtat ctctggccac catttgctgg   6720 

gccgtggctg tggagggcag ctggtgtttc tgtttctttc tgggcacgct gcctggctgg   6780 

ccagtctcag aggccacatg tatttttcct catagtctga aggagacaga taaactgaag   6840 

cttcaggttg gagggcagtg atgggcaagt gctatacaga gccttctggg tctgataagc   6900 

ccacagagag ctttgttttc cttctcaaat ttcttttttt aaaaggcaga atgtcgccca   6960 

gacttgtctc caactcctgc tcaacaatac ctccttgctg ggccgtggtg ggacaccttt   7020 

aatccaagaa ctcaggagac agaggccagt ggatctctga gttccagcca gggctgtaca   7080 

gagaaaccct acaacaaaca aacaaacaaa acaaaacaaa agagtacttc ctgcctcgtc   7140 

ctcctaagta ctgggactac agagtgtatc gtttatttta attaactcat gtcgtattac   7200 

aataattaga gactagatta ttacttcctt cttcagaaag gtacattggg cagagagggg   7260 

ctaatttact tacccagggt ctcaaaatca ggtggaaaac tcccagttta actgtaccac   7320 

ctgattctca ggctgcgctc tgcttcccaa gggaggtcca tctgtggagc ccaatagtcc   7380 

tcgggggtaa ggaacagaga ggatgcccac ggtgttgttt gcttttttaa cactagggaa   7440 

aaccccggcc tagtgtttgt tccatgtgca ttctgccact gagtcagaca tgcacagccc   7500 

cttcctgtgg actcttcccc ctagcaggta gagggagaca gggcagctag gtgtatgaat   7560 

ggggaagctg gaactttagt gccagggacc tttatggtgg ggtttccccc acgaaccatc   7620 

ctggcagatg tccacagcag atgtgtctcc agttcactgt gtcttactct ctgactcttc   7680 

tccctcgact ttcgctggtc caaacaggga tatttccgac aaaagggtgg tagcatctac   7740 

cctgagctaa acaagatgaa aggcaaccat ttctagaggt gctgccatct tgaaaattga   7800 

gttcttagtt ggctttatgg gcatttatcc tcacagacat gttagccttc caaaaacatt   7860 

caaacaaaac caagtgaaat caagggaaca gaaaacagag gacaagtgtt ttgtgctctc   7920 

ttctcttctc ccacccctct ccctctccct ctccccctcc ccacctcccc ctctctgtct   7980 

ctgttggtgt caagtgactt cctcagtcat tctctacatt tccctgtgtg tgacaggact   8040 

cttcactcac cgatttagta gactggctgg ccagtgggct ctagggatac tccagtctct   8100 

gcctccccag cactcggatt ctaggctcag agcactacac tagccttccc atggtcctcg   8160 

tgatcccagc tcagacccct atgcttatat aggcctggag tttacagact gagccatatc   8220 

ctagccctgg tttgccttaa gttacccttc ttccccagta atgcaaacag acattaggaa   8280 

gtacttagga gccaggtgtt tccctactgg cccctggatc ggcctaagaa gggcagtgtg   8340 

ctttctggca ctatgcctgg aagggtgagg atagctaaac cctggcccag gactgggctg   8400 

tgtggaagaa ggcagccaaa tgtagagaga gtttgcctat ctgtgtgtcg tgagacacag   8460 

gacagatgct tttttgcagt ttcctgcata gtttctctag tctggaggga tctcctggcc   8520 

catagtgggt ctactgtcac catgatggcc acagccaggg aaggcctgta ctgccttagg   8580 

ctactgttcc ctccttcagt gacaaacctt ctttgttttt gatttttttg ttttgttttg   8640 

ttttgttttt ttggttttcg agacagagtt tctctgtata gccctggctg tcctggaact   8700 

cactttgtag accaggctgg ccgcgcctag ttttgttttt gcttgttttg ttttgtttta   8760 

tgaggcaggg tctcacatat acctgaggct ggtttttgtc tcactatata cctgaggcta   8820 

gccttgaaca cttgattctc ctgcttccag cttcccaagt gccaggatta caggcttcaa   8880 

atctttcttc agaggcagta aaagaacagc tgaagcctgg gtactcgaga ttccagcttg   8940 

tgtgatccag agcccttggc tgtaggcttt tacctgagcc agcagtttag ttttcataac   9000 

tggtgtatgc atacatgttt ctcctgtagt ggtgctgttc ccaataagta cgttacctca   9060 

gcccacctta tgtgtcctca gaacagacag ctagccttcc aaggacaagt gtgactgatg   9120 

ggggaaaagg gaccctggaa ctcaccagag ccaccctcct ctagctgagg acatagaaaa   9180 

cctttacctg gatttctgtg ggaacttccc aacaggcttt tcctaaccag tcttggaaag   9240 

gtgtattgag actgggtgac accatctgga agaggccttg gaacccatag gagcctacca   9300 

tgcctcctca gtctggcgtg ttgctatctt atagcataga cctatcttcc cttgagttct   9360 

agacaaggca agtttctggc caggacatgg tcttgttttt ctttgagcat cttctagaaa   9420 

ccagggagac cataccacaa agcccttact ggactgacta ctgcatgcgc acctccagga   9480 

gcccatctca tcaggcaagg tgactgctgt cctgtctctc tgatggaggc cattgcccct   9540 

ttaacaaacg aataaaggtc gctctcccct ctagggtgtg gaagacagga aatggctgtt   9600 

acccaatgca ggcccactgc cagctctgcc ctcagagcac ggtgcagaca gtccagtcgt   9660 

cctccattgg attctctgct gggctaggca cccccagtcc ctctgtggct aagctaagaa   9720 

aaagagagag aaaaaaaaaa aaaaaagagt aaagcattgg gggtggggca ggaagagagc   9780 

acaggcgtgc aaacatcgaa gagcggcctc tgtgacatct gtctgcgccc ctgttggctc   9840 

acccttagga catctgactc cctttctgct agccatcttg tcccacccaa tgcttagata   9900 

tttcagaagc ctcggtcctg ggtagggagg gaaagcaggt ctctgtatct tataggcctc   9960 

agacaaccag gacagccatc ttctgcaggc ctagtgaggc cccagggatg ggcagcttca  10020 

gtggcatggt gcacacgccc ttttccacac caccctttgg caagattact ttctgtgcta  10080 

atggttaaag gcagaaacct ttgcccacta agcagttgct gcgcccctga gctacgctcg  10140 

cgttcttaaa accattgtat tgctggtgtg gtgggtcaag tctgtgatgc cagcacttgg  10200 

caggccaagg caggaatgag aaggagaaca agtttcaaag caagcctggg cttcatagta  10260 

agaacttgtc tccaaagccc aaagaaaggg ctggagatac aggacagctg gcagaaacca  10320 

ggcacagagg ttggcatctg tagacccaac acccggacag tggaggcagg aggatcagaa  10380 

gggaagaccg ttcttgcctg aacgtcaagt tctaggccag gctgagggcc atgccaggct  10440 

ctctactgtc tatgtatgtg ggtgtttgct tacaccactt tcaaacctgg tgcccaagga  10500 

ggccagaaga tggggtcgaa tcccctggaa ctagagttac agacaaatat gagctgctgt  10560 

gtaggttctg ggaaatgaac ccaggtcctc tggaagagca gcctgtgctt ttaacaactg  10620 

ggccattttt ccggcccata ttcattttta ttacgtgtag ttgtttattt cattatggga  10680 

catcccacag catgcacctg gctatcagac ttgcggaagt cagttctttc tttgcccagt  10740 

gtgggtccta aggcttcatt cagttcatgt tggcaggctt gtgccccctg ctttagatgc  10800 

cacgtcatct ccagccactc acatattctt gctacccgtt ccttgtcaga tactttgtag  10860 

acgtttcctc cccaggctgg atttgaactc actgtgcaga tccgtctgtc ctgttttagc  10920 

ttcctggatg ttgagattac agataggaag caccatgtct gactcggttt tatcgtctca  10980 

ggagtgtctt ttgaatcata aaagttttca actttgaaga ctacgttagg taattttttt  11040 

ttcttttgtt acttgtgcct ctgggctatg tctaagacgt tgcctaatac aagataattg  11100 

agacttcctc tcgtgttctc tttttaaatt ttttatttta taaattatgt gtatcggtgt  11160 

tttgactgcg tgtctgtgta ctatgtctgt gtctggtgcc ccaagaggcc cagaaaagga  11220 

cattgggtct cctgagcctg gagtttcagt tctgagccag tggatcctgg gaatcaaacc  11280 

caggtcctct ggaagagtag ccagtactct actgctgaac cagctactct ccagccccca  11340 

cccttcttac acttaggtct atctgttttg gtttggtttg gtttttaaga atttgttatt  11400 

caggggcaag agagatggct cagcagttaa gagcactgac tgctcttcca gaggtcctga  11460 

gttcaattcc cagcaaccac atggtggctg aaatgggatc tgataccctc ttctagtgtg  11520 

tctgaagaca gtgacagtat actaatacat caaataaata aataaataaa tctttttaaa  11580 

aaataaaaag agaatttgtt attcaaagcc aggtgcatct ctttgggagg ctagcctagt  11640 

ctacatagta agtttgagaa cagtcagggc tacatagtga gacctatctc aaaagaaaat  11700 

ctgttattca gactggagag atggtgactc agtggttaag agcactggct gctcttcccg  11760 

aggacttgtg tgactcctgg catccacatg ggagcacgcc accctctgta actccagttc  11820 

caggtcatct ggcaccctct tctggcctcc acgggcacca ggcacagaga tacatgcagg  11880 

caaaacacca tatacatcaa ataaaaataa aatagtttgt tatctttttt tttgaaaggg  11940 

aagacaaagt tttactttta aaaaagatta caagcacccc aaataacatg taacgagttg  12000 

agtcctcgca tctcgtgatt tgggatagga tacactaaca gcagccggaa taagcatacc  12060 

atattgactg tcctaaatta tccaggctag agtactgtaa ggctggctgc tacttcatag  12120 

gagttgctaa tagctattac tacttttcca taaataacgc ccctgacctt taagaaagta  12180 

gaagggaaca gcttactccc tttctttcaa agaatttttt ctacttgact aataaaaaag  12240 

tcagcactga tatccattac ttgcagaaga cacaggaaac aggtgacaaa cactccttaa  12300 

agacacacaa gataagaaga tggaacttca ggtacatagc aagtcggtac aaaaagctag  12360 

atttgatact cttaaaacgt gaagggtcct acaacggcat agagaaataa tttaatgcct  12420 

tccagaacag aactcgagct ctgtggaggt ttcctattct ataggggcag atctcatgcc  12480 

aacccacaga gcaggcgctt ccacctccta tccctttatg cggtagcttt catggatttc  12540 

tggctggatg tcacacacag aggccaagag gtcattcagg actccatccc tgttctgctc  12600 

gaagtggttc tggaggacgt tcatcttccc ctgggtctcc tcttccacct cactgctgca  12660 

gctgccatga gaccccagtg ctgcagcttc cttggcctct gcagtactgt tcaatttcag  12720 

cctggcggct tctttggcct gcttcggcct ccggttcttt cacttgcagg ccttggacac  12780 

cttcttggct gcctgcagta gctgctggat gccccgcaac tgactctcgc cattgctgag  12840 

gggactttgg gccgagagaa tggcctaaat caaccaacgg ctcaaacata gtcagaagcc  12900 

cctccgtttg atgtcattta atgagccttt ctgtgtagct tcaggtcact ccctgaggcc  12960 

tggaacaccc tgaatctttt tcagcttttc tgctgaattt ggctgtcacc aggacagact  13020 

gctgagggag tgtgttagta ctccagagga gcccagttgt cactatgact ggagcagcgc  13080 

agtcttgttt gtggcactgt tgggctatgt ctgctcactg acagttggga tcagttcctc  13140 

ttaggtgact cataactgtt gcggtaaatc tcctcccaaa tatgccccgg caatgaaaac  13200 

acaacacagt tcatatgaat acatgctgtg cgcctagatt gggcagatct accgctacac  13260 

taccatcttc cacatctatg agacccctta gaacttgcgg tttctccagg ccttgtgctt  13320 

ctgctccact tttccccttc tttctccttg tctgtgtcct ctccctcttc cattttctct  13380 

ttgttctctc cccccacctt ccgctccacc ttccctttta tctgcccaaa cttcagctcc  13440 

cctttatttt acaaattaag gtgggaagca ggtttacagg aaatcacctg agtgctgact  13500 

atgttcttgt tcacaaccac tctcaggaga acggaattaa catcaaatat aattagcccc  13560 

agggctatct gcaacacata acaactatgt cagtgtgatc tggctctatc tgcaagagtt  13620 

gaccctctgg tgatgccctg actgagcgtg tcctgcgctt gctaatgctg tggtgctgcc  13680 

cctggatggt atgtccacgg ccaacatatg tccaaaagga aagcccctgt cagctgttgt  13740 

ttttttcaaa tttatgtcta tgtgtgtgag tattttccct ttttgtatat ctgtgtacca  13800 

catgggtgcc cagtgcctgt ggaggcagat gccccagtac tggtgttaca ggcagttaat  13860 

atgagctggg aattgaaccc aggtcctttg gaagagcagc cagtgctctt aacttctgag  13920 

tcctctctgc agccttctta gcatccattt ttaatctttt gtatgacatc tggcagaggt  13980 

aggaattcat gccttttggg gggatagttg gctatcccag tgtccttggt taaaactgtc  14040 

tgttctttcc ctggcggtgg cccgggtcag tgtctgatga actcgatgct cactgctctc  14100 

tgatttcttc aaccaggccc gcaccttcat gacgtcatga cgagagctat gggaaggttt  14160 

gaaatcagga agtacaagtc tgtcatccac tttgttcctt ttcaagaatg gcgatttttg  14220 

aaaatgtcct ccgcgttcat gtatggattt aggaattgtt tgtcactttc tggagtattt  14280 

tttataggaa ttgtgtggag tgctgtagtc tgatagtgtg ttgtctcttc cagcccctga  14340 

caggtgcttg ccttccgttg tttatctcaa caagttttgc agttttcgtt tagtgtctaa  14400 

tgctcgtata acattcgctc ctaaatgctt tgtgcattaa ttttgttcac ggcactgggg  14460 

ttgctctcaa gctctcggta gacgtgtgtt ctactgtgga gatgcaggcc gggtcttagg  14520 

attttctgtc tcttggtagc acaataatca tttcatttta ttttgggtta tgagtagtgt  14580 

atagaaaaac aggacagcag gggcttgctc tctgctactt tgttttcttc atgaattcct  14640 

tgggtgctgt gtgtaaggtc atgtcagatc actgtgttca ggggcttcca gaagattcca  14700 

ctgtgcagct aagcttgaaa attgctgagg aagctgggca ccacagcacc tacctgtctt  14760 

cctgaggcct gcaaggtagc gccaagagta gacctcgctg gcggcgtgcc tggcaccccc  14820 

cgcctgccat ggaacttgtc ttggtctatg attggtacat gatagacaaa gaggctcttt  14880 

tttgtcacat caaggattca gctttgtgac cttaacgttt gttcatcttt atgaataggt  14940 

gacatagctg ctttctgttg gggggctggg agagcacacc cggttgctgg actgttttct  15000 

ctgcgtcctt ggtcgcaagc tcggttgaac tgttttgtgt ccaaggagaa gaacagcatc  15060 

cgttactgga cctgtgagtt tgggtctctt tgtcctgcct ccctctccct gcctgcctat  15120 

gtgtgctcgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtagaggg  15180 

aacctcaatt gagaaaatgc ctccatcaga tttgcttgta ggtaagcccg cagggtattt  15240 

tcttgattgg tgatggtgtg ggaggtctgg cttactgtgg acagtgccgc tcctgggcag  15300 

gtggccctga gttctgtaag aaagagcctg agcaagacat ggaacaagtc agtaagcggc  15360 

ccccctccct ggccatgact ccagctcctg cctccaggtt cctgccttga cttccctcag  15420 

ggggaggggg acggggacgg gacctgagag ttgttgtgct gagatatgta cttttctccc  15480 

caaattgttt ttggtcaagt gttttattac gatagaaagt aaactaaaac acactctccc  15540 

cacacacaca ctgactccac cccacacacc gtgaacacag ggccttgagg attccagaca  15600 

gccttgtttt gtatttattt tgggacaagg tcttagaaag ttgaacttgt gatcctcctg  15660 

cctcagcctt ttgagtagct gggattataa tctgtgtcac cgagtttgtt cttgacctaa  15720 

gtagttgaga agagcctttg ctcttgtgta aatgggaaaa ggtgctttag tcacagaggt  15780 

ttaggctctg gcttctcact gatgcagcac caactggagg agacattcat acaaattaaa  15840 

catttttagg atttttaaaa agtgtgtttc aatgttacat ttggggtaag aatgaaaata  15900 

caggaattat gtcggtgcat tgggtgtttt agattgtgtg tgtgtgtgtg tgtgtgtgtg  15960 

tgtgtgtgtg tgtgtgcgtg cacagagttt tgaactgaag gttttgctca tgctaagcat  16020 

gtgtgctatc acccagttcc tctgaaaaag catctctaat agaaactgcc cattctcggg  16080 

cactccgggt agcagagcag cttccgctac tgcgtgttga ctttattgtg ctcttggctt  16140 

tttagacatt gtgggaaggg gtggacaaag ctcactgttt atgaaacagt ctgggtttgt  16200 

gtcattaatg gataaccatg cctattctcg tgcatgtgac cctgtgttaa ttggatgtcc  16260 

taccacctaa tgcttcttac aacacttgat gtttactgtt tccaaaattg gacctagatt  16320 

tagaaaaaac aaaacaaaac aaaacaaaac aaaacaaaac ttgatttgct tatttctatt  16380 

ttgcatgctg gggatggaat gctcaggcct tactcttgca ggcaggcatt ctaccatcaa  16440 

gctgtgttcc cagccctttc aggagcctga cacctaaagc tgagcttggg caatcctgga  16500 

aaatctcagg tgtggccatt tgtattgtaa aaagggaaaa ttagggagag atggagggat  16560 

ggatactgga aactgaactc atgtcctctg gtaggataga cagaacactt aaccactgag  16620 

ccttctgcaa ccccctttag agagagagag ggagagagag agagagagag agagagagag  16680 

agagagagag agagagcgtg catgtgtgtg ttacacacag aggccagaac agctgtcctg  16740 

gaactcactt tgtagaccag gctggcctcg aactcagaaa tccgcctgca tctgcctccc  16800 

gagttctggg attaaaggcg tgcgccacca cggcccagct ttcaagacaa attcttaacc  16860 

gccagtccat ctcgccattc tccaaccagt cccttaaaaa tatttttttt tcaggtgttg  16920 

agggtctagc cccgggatac aggcatacta ggcacggctg aagcactgag ctccacacca  16980 

caattgggta ttattaccgt cttaccctct aggttattga tatgctgcag aatacagata  17040 

ttaatgcagg cacttgtcca caggcctttg tccagtgcag tgtggttatt atcttacagc  17100 

tattggcagt cttgcctgcg tctctaagtt cttctgtttc tcatcatctg tgcatatggt  17160 

tctttgtcat ttgagttttg tttatttact tatttgtttg tttattttta tggagacaag  17220 

gtattgtata gcccagcctg gcttccagct cacagtgttg aagaaggcgg ccgggaactt  17280 

ctgcttcctg cgtgctgcag ttacgggtgt gtgccatcgt ctccggcagc ccggggctct  17340 

gcatgcatgt gaggcaggca ctctaccaac agggctgcat ctcaagcacc tgggcagttt  17400 

tagcacagtt ccttggtttc ccattaagta atgagttaaa tatttaacat atgtccattt  17460 

gaaaagatgg aaaacaactt ctcctggtca ctcggcattc atcagccaga agtctgggag  17520 

gctttttctt ctctggatct ccacttggcg gcgttctctg cctgctctgt agcctttgat  17580 

aagtggatgg ctgggtgccc tctccgtaat atttatcaca tttttctcgg ttacttgtat  17640 

agataaacct cagcagggca ggggcacaag gacacccagc tctgtgtaac agtactttgt  17700 

accttcctcc ctattggtgt gtcccgagtc tgcacttcgg gtgggcgggg ttttgtgaag  17760 

ttcagagttt tcagctactt cagggctttt ggcttctaca gtacaagaga aacttccagg  17820 

ttcctgggag agtgagttgg agtctgagta gtgtgaccca cgtgagctgc tgtccattcc  17880 

tcttactcag gacacagctc tctgctcaga aatagctctc tcgtcccaag actccacctg  17940 

gtggcttctg gaagaagtgg cctctgtgat ggtggagatt gacagctctg actgtgattg  18000 

acagctctga ccaccatgag gtgcatgcaa agtgctttca cacctgtcta ataattctgg  18060 

atgtaatgag aaataccaag caaggtgttt ttttttttaa ttagaatttt tattcatcac  18120 

tgtgtgtata tgagggaggt gaactcatgc gtatggaggg aagagggacc tggaaccggc  18180 

tcctctttga cctttcacat tgttccaggg atggaatgca ggccatctgg cttgctgact  18240 

ggcacattca ccagctctct tgcttgcatc tgatcttagc ttttttgagg gacctctaca  18300 

ctattttcca tagtagccat attaatttgc attctcagta acagtatata caatgaatgg  18360 

atatactttt ttaaccatgc aacaaaacct ttattaacat tttaaacaga tgttccgcta  18420 

ttactgaaac tttgtggggg ttggggcggg ggcaggtttc aagacagggt ttttctctga  18480 

atagtcctgg ctgccctgaa acttggtttg tagaccaggc tagccgaaaa ctcagggatc  18540 

cacctccttc tgcctccagg tgctggaatt aaagttctat accaccaagc ctggctgtac  18600 

tgaaacttat aatttctaaa ttcaaatgca caaatggttt tagtgtagag taataccatt  18660 

agtgcctacg ggaaatttag gctgaagaac ggagaccatg tgtgggcttg agtcttttct  18720 

ggatcaaaaa gagtatggtc atctttcagc tgcttgcctg taacgatgag cgtctgctgg  18780 

gtggggtggg aggtgccctc ctaatcctgg gtcttaccct tcacattctc tgtggtatca  18840 

gtgggctcta cctcagggtc tgggtcttca caaagattca catctttttt gggggagggg  18900 

gtgcgttgag acagcgtttc tctgtgtagt cctggctgtc ctggaactca ctttgtagac  18960 

caggctggcc ttgaactcag aaatctgcct gcctttgtct cctgagtgct gggattaaag  19020 

gcgtgtgcca tcatgcccgg caagactcac atcttaacct gttaatgaag ggattaaagt  19080 

gcaaagttca aagcacatca gggcacctag ttataagagc ctctgcactg gacaaagctg  19140 

ctcgtctgga catcctcaat gaagttcttc aatgactttg gtccagtcag ctatggtaga  19200 

tcagaagact tgcatggcgg gcacgtttta ccagccaagc tgccttgccg gctcctccag  19260 

atgacatctt cttcccatta agttggaata catactgtgt gctttgcctc atcgtgtgga  19320 

aagaggaagt ggttggtggt ttgggggcac tgtggtcctg tagtgtagat gccctgcagt  19380 

cttgcaggag tgtgtgacta gctgggaaac ccactaacca gtgtgaggat tagcagcagc  19440 

agttcttgtg ggaagcgccg gttggcctga tcagacttac tgaacatggg aagaaagctg  19500 

agctctggag aactggcctg gggatgccca ggtcagtgcc agcggaggct tcaaggagga  19560 

agactgcaga cctgactcac tgggtctgtg tggagagcaa acaaatgagc caaagccagc  19620 

ggtgtggctg ggtgtgcctc agctgcaggt gtgacagtgt cctgtatccc gcggggcccc  19680 

gcagaggcat tgctttaggg aacagccacc catggcttgt atatgtcctt tttcaggtga  19740 

ttccctggac tctgtgagct ggcagtgctt ggagctacac agcttgtgcc atg gac     19796 
                                                        Met Asp 
                                                         1 

tct gag gttagattct ggtatctttt cattttgttc atcctgggtg tccccgttaa     19852 
Ser Glu 

gcaacctgac ccctcagttg tcaggtctgg caaggtgtac ctcagataat ccaacagagt  19912 

tcatctccac tggcacctga tagggactta gtacagaatg gggaaggggg acgtccttcc  19972 

agaaggacgg aacggcgtga ctgtcagctt ggtagacata gcaagggcgg cacaaaggcg  20032 

ggacagaaaa gatctggaag gttccctttt gccccagtca gggggctgag ctgggctcgg  20092 

gcaatagtgc tttctagcct cccagtatct cctgctgtcc tgcagggcct cttgagagtg  20152 

ggcccctcct ggacaacggt agacttgctg ctgtcccctt cttctacctt ggagcaggaa  20212 

agctgaggca cagaagaaag tgaaatgctg acattttctc ttacatcttg gcatttgaca  20272 

tccttgcccc acatcagaac ttgtatctta ttgtagatgt ttctgacttt atgacaactg  20332 

ttatgcacac agttgaggga cattaagtga agcaggtttt gctactacgt ttttttgtac  20392 

tacagggact catggaacag gcgttgctga gtgctcctcc ttttttttgt tttttgtttt  20452 

tttcgagaca ggatttctct gtatagccct ggctgtcctg gaactcactg tgtagaccag  20512 

gctggcttcg aactcagaaa tccgcctgcc tctgcctctg cctcccgagt gctgggatta  20572 

aaggcgtgcg ccaccacgcc tggcgctaag tgctccttca tagtgctcct acccagggct  20632 

gcttttgtac acaccataga actggcagag aggccggtga gcaagaccct ccctgctgcc  20692 

tctgatagtg cacatgtccc cctgaaaggc acaggcagag tcggacctgg gtccctgctt  20752 

cctagagttt atcaggcatc ctgtgtctgc tcatgaggga gtgaggggaa agaggaaccg  20812 

cttgctgcta ggagcacagc ccgtacagtc aggctcagcc ctgaacggaa acatggatgg  20872 

aactgaagta gtgacatttg cctgccaccc cagtgtccct gagaccttcc ctcgaagcag  20932 

cttccccagt gggtgtcttc aggaggggat ctgtagaagg tggctcgatg gccccttggt  20992 

gtcttctgtt tggcaagcac accacagcct gtttctctgc ccctgggcct ctcactaggg  21052 

catttagatc ctccgagtta ttgattgtca caggccattg tgactcgggt ccaactgtgc  21112 

tctgacccag gctcccgtga gccttcctga ctccccttcc accttaggtc agcaacggtt  21172 

ccggcctggg ggccaagcac atcacagacc tgctggtgtt cggctttctc caaagctctg  21232 

gctgtactcg ccaagagctg gaggtgctgg gtcgggaact gcctgtgcaa gcttactggg  21292 

aggcagacct cgaagacgag ctgcagacag acggcagcca ggccagccgc tccttcaacc  21352 

aaggaagaat agagccaggt aggtcctggc cttgtccacc tcatcccaaa tgtagccttt  21412 

actgaccccc aaaagctaca agggcttttg gagctcagtc tctaacctta cattgtcagg  21472 

ctggtgtgtg tgtgcatgtc atgtgactcc tgccttgtga tctgcatgtg actgccccca  21532 

gtaatgtcca gttcatatga catcgcctgt atcaggacaa ctaattagaa agttcttcct  21592 

tctgatgagt cctgagttct cttcaggtct ggacctgagg atcctctctg gaccaatatt  21652 

taaaacatgg tttttaaaac atatgtccca aacagttata gtacagccaa agtatggaaa  21712 

ttgattgtct agtttaggct tcattgctgt gaaaagacac catgaccaag gcaactgttt  21772 

tttgaggggg agggggcttc gagacagggt gtctctgtgt agccctggct atcctggaac  21832 

tcactctgta gaccaggctg gcctcgaact cagagatccg cctgcctctg cctcccatgt  21892 

gctggctagg ttttttattt ttttattttt ttttatttct tagttctttc ctgcaactat  21952 

caagtcattc agaaaagagg agtcaagaga ggggatgagg tacatttgaa ataaaaaact  22012 

ataatgatga ttggtcctgc ttctgcctcc ctagtgctgg gattaaaggt gtgcgccacc  22072 

acgcccagcc caaggcaatt cttataaagg acaaatttgg ttgaggctgg cttacaagtt  22132 

cagaaggtca gtccattaac atcatggcag gaagcatggc agcgtccagg taggatggtg  22192 

ctggaggaag agctgagagc tctgcatctt gatccagctg tcatcttccg ggctgctagg  22252 

aggagggtct gaaagcccac tcccacactt cttccaacaa ggacacactt cctatcagtg  22312 

ccactatctg ggccaagcat gttcaagcca ccatgctggt caagatgtta taacccagaa  22372 

gtgccatcag cttcagcttg tggagttttg gaaagtagca aggcagagtc cttcgtcctg  22432 

ccattcagat ctgggaggtc tgggacattg ctagtctggt catggctgcc aggtaagcat  22492 

ccttcaatag ccacacagca cctcatttgt gtaggctagc tgaactctca atccagtgaa  22552 

aactcctgcc gttagagtca ttttgcctcc taaatgaaac tttaacatat gtgacttgct  22612 

attacctaaa gagatgaccg agtattgaag tatcctgacc ctcatttcca gataaggaaa  22672 

ctgaggcaca gcagagaaat ggctgacctc agatcaaact gcccatgcag caggagcaag  22732 

gctcaaccaa gctgctcctt catcagtgca gtcacctcct gctaagcctg tgtcactcgg  22792 

ctgctcctag ccttcacctg tcccctgtcc cctgtcccca tgctgtgttt acagcaactg  22852 

aggagacctc cctaaaggct gaggtgcagc gagtgctcag agcgctgtgg gcagcatgca  22912 

ggtgggcatc actgagttct tcagagtgta caggcctggc tcgggctctg ctcctccagc  22972 

aggttctgga gctgcatgat tttttttaaa atgcttgtct gtctgtctgt ctgtctgtct  23032 

gtctgagtat ggggtatgca catgccctag catatgtatg gagtcagagc tggctgtttt  23092 

ccttccacca tgtgtgtcct gggatcaaac tcaggtcagg atacttcagg actctaagca  23152 

ctgctgcctc cgatcttgga cacagaggct tcactgccct ctagtggttg caagggagac  23212 

cagcagctag tttggcttcc ctacccccct ctggctagtt tatttctttt gagacagggc  23272 

cttaccctgc ctagcctgaa atttgttgtg ttgaccaggc taatcatgaa ctcccagaac  23332 

tctgcctgct tctgccaaat gtggttcatt tttcaaatgc cctgaagtgg tatcttgagt  23392 

aggctgggat gtgacaggta ttctctacaa gctgggtttt accatagcct tgtctccgaa  23452 

gcccaccagt gagccagcca gccaggccaa aactgaagag aagcgccagg cagtccagga  23512 

aaggctcagg aagttcaggg cagcgggagg aggctctggc tgtgcgcagg tgtctgtcac  23572 

tctgtgccat acccgcttct ttctgcatca gtccatgcca gacttcaaag cctggcttaa  23632 

gtcacgagac tggggatgac gaggctttgc agacgatcga tcggctgcag attgggagca  23692 

gggcaaagta gtggcttcag caagccagtg agcagctgag tctgcctaga acactcggct  23752 

agtagtggat ttaaatcaca gggaaccgga agccatgcag ttactgtcac ctaagcagaa  23812 

gcagtgagca ccagagaggc cttgaggaga gcagtgtggt gaccatgtga caggcatgga  23872 

ctgagggagg gcctggagta ccgctgaatg ctgaagcagt tgcccactgc attaaagcag  23932 

cagtgacaca ggcaggacac aggacaggag cacccccaac cccccagccc ccgcagcagc  23992 

aagcatataa tctgggacag gcctgcttct ccagccaggt tctgctaccc aggccttccc  24052 

tgcacccggg gaggggcggc actcatggtc ctcactaggg caggtgcgga ggtaggaagt  24112 

ggcctgaagc tgttgacaga accattgctg agtcttgtat ttgttgccta aacagattct  24172 

gaaagtcagg aagaaatcat ccacaacatt gccagacatc tcgcccaaat aggcgatgag  24232 

atggaccaca acatccagcc cacactggtg agacagctag ccgcacagtt catgaatggc  24292 

agcctgtcgg aggaagtaag tatgactctg gtctgggagc ccctcttatg ggacatttcg  24352 

gaagtgtggg acatttttcc ttgtcgaacc agtctttccc aggaagtaaa ccctgtcctt  24412 

gactgcccgt cagcatggtc tctccaaaga atttagtcag agtacagagc ttaggagtca  24472 

ggcctccagg aagatccctg aagtacctga tctgtacaga tactcagtct tctcttgtgg  24532 

cgaactccat gtcgttcccc cagggtgagc atctgctcgg ctgtgtggtt agaatcagca  24592 

catggaaacc gatacaagtc cacctcttgc tgggtatacg gtgaaggacc caaagctcgt  24652 

tcctcagcac cgggtccttc ctaaagcaga ggtggagggg tggtggggag aggggagaga  24712 

gagaaaccaa accccggggc tgtgaagtac ctgcccaagg aggaagattc tgttcttagg  24772 

acttccagca gctgaaatcg tggctgccct caccatctag attcattgtg cctacataca  24832 

gcctgtcttt gctggcactc tctctacctg ccactctcca gtggctgtca aagacacaca  24892 

catttgtcaa cagccttggg ctcctcctat ggggtagatt ctttaatgtg agccacagaa  24952 

cctgaagctc actttccacc ccaccttgtt tttttgtttt ttgttttttt gttttttttt  25012 

tgaggcaggg tttctctgta tagccctggc tgtcctggaa ctcactttgt agaccaggct  25072 

ggccttgaac tcagaaatcc acctgcctct gcctcccgag tgctgggatt aaaggcctgc  25132 

actcccctcc ccatttttta aagagttaac gttacctgtt tctgcgtgca cctcatgtgt  25192 

gagtacatga gcatgcttgc aggtacatgc attgccatca gatcccctgg agctgaagtt  25252 

tcaggccatt gtgagctgtt gcctataggt gctgggaact gaacgggctc ctctggcaga  25312 

gcagtacatg ctcttcaggt ccaggggtcc agtatcttcc tttcctgcct gaagggaaga  25372 

taacatgtag cccctaaagc taagctcaca gtaacatgag cctaagatgt gctcgtgtcc  25432 

agccaattct gtaagcatct gagtgcaggg aagagctcag acgcccatat gtcagtagtg  25492 

tgtacaggct actcactaac catgcactgg tgagtctcca cgtccctctc tggtctgtgg  25552 

agagtgaatc ctctatcatt tcctccaccc aacgttctta gctatttaac caccactccc  25612 

ctctgaaagg ctgcttcctc ctttggcctg atttggtctc tctgaaggaa gagcatcagt  25672 

aaactgtctt ctttaatgta caggacaaaa ggaactgcct ggccaaagcc cttgatgagg  25732 

tgaagacagc cttccccaga gacatggaga acgacaaggc catgctgata atgacaatgc  25792 

tgttggccaa aaaagtggcc agtcacgcac catctttgct ccgtgatgtc ttccacacga  25852 

ctgtcaactt tattaaccag aacctattct cctatgtgag gaacttggtt agaaacgtaa  25912 

gagccagcag tgacaccagc ccctgcctgc ttgcctaccc tattctaatg cagcagagcc  25972 

tctgctgaag cccctctggc ccgctctccc ttttgaccac ccgcagactg agagaggcaa  26032 

ggctgtttca caccactgat gggaatcgag caagctgggg ggacgtggag tgtttaggaa  26092 

gatgactaag ggctcagccc cctaagtgtg tgtggtgtgc acatggaagc cagaggtcat  26152 

tattgggtgc ctttttatct cgctctacct atctttgtga ggtagggttg gttctccgtg  26212 

aagtcagaac ttgccggtta ggctaaacta gcaaaccctg ggcttccact gcctgccttc  26272 

ccttccctca ctggggtacc agttgtttaa tgtgtattga tgctctacct gaatgtgtgc  26332 

ctgtggacca tgtgtgcctg atgcctggat agccaggagg gtgctgcatc atctgggatt  26392 

gagttgcaag tggttgtgag ctgccatatg ggtgccagga atctgaactc ggttcttcag  26452 

gcctctgtag ctcttactga gccatctgca cagccccagg tattatgagt aatcagaaag  26512 

tgactacact tatttgtgtg cgcatgttgc tgtgggagca tgtgtgctac agcatagtcg  26572 

gtcaggacga ctctgaggtc ccagggattg cactcagctc atcaggcttg gcactgtaag  26632 

ccatggccca tgacttagat tctttcgaag ggcgcttccc gaggatggag agagaaactg  26692 

ataggagtaa taaatgagtt aagtgagaat cgctgtcaag ctctccagta agcctgagga  26752 

cgggcccatt gctagggtag ccctgagttt ctattgcgca tgctcaggaa gtggttacac  26812 

ggagctaagc ccaaggtcag tctactgaga ctgctggaaa atgaccacgt gttcttagag  26872 

tcttgtgctc tggttacaca aacccaagtg ggagctggat ggagatacct aacctgcact  26932 

aggattttac aatgtttggg attttagaac ctgtcagaaa cattatccga gattcttttg  26992 

gggggagggg gtttttgttt attctgggtg aaggcagagt ccacattccc agatggcaat  27052 

ggaatgcaag gcaatcctcc tgcctcagca tctgcaggca tgcaccccca cacctgggtg  27112 

ggtggagcag aggacaggtc tctgtgtgcc aggcaggcac tgttgactga gcagcagccc  27172 

agtgcttgtt ttctaacgca ccgtatcctc caatgagact tactctgctg cctctttctt  27232 

aggagatgga ctgaggagcc cgcacaagcc cgatggtgac actgcctcca gaggaaccgc  27292 

gaccatggaa agaccttggc ctgaagacag gtcccagaga acagctgtct ccctatttcc  27352 

aggtggtggg aaccccaagc tggtgattca ctggacatct ctgcgttcag cttgagtgta  27412 

tctgaagagt ttacgccggc tcctgcatcc acaccatgta cctttgtcct atcagctgta  27472 

tgggttccca cttgggaatg aaacttaaca gcaggctgta aggcagaaaa gcatctttgt  27532 

aatgccaagt gactgttcct gagagccagc tctgggctgt cttcaccatg taggtgggct  27592 

tctgtctaag gagaacagca ttaggagagg tgcatcggcc catgagcgtg aagtccaccc  27652 

agcctagtgg acactgaagt gctcacaagg cctccacctg cctttgtaaa agccgaatgg  27712 

ctgatctcaa accatgggaa gcccgaccgc cccacccctc ctcaccccag cgtttagctg  27772 

tttcaggggt cagctattat ctcaagattt ctatccaagt ggaaacaaac tgaatcatgc  27832 

acacgactta tctgtgtggt gtcagttaca ctcaggctct tgctacggaa tgcaaagaac  27892 

aactcacata ccagtgtcaa acagaatgca cagaagagac ctaaaacagc agcaggtcac  27952 

tcggttcaca aaaggtgact cccagtcagg tctgacactg tcttggttgt agagcacagc  28012 

tgccatcctc tttccctggg taacatcaca gaagattcca tatcaaaagc aaatgttccc  28072 

tccgcttctg tatttcagag acaaggcctc actgtatcct caagcgttgt tacgtcttgt  28132 

gctgaacttt gcttaaagct gggatcgtca gcacgagccg ccacagcctg caagtattct  28192 

agttctgaac tcatcccagc catggtggct gtgatggctt gggtgtatca tacctgtaaa  28252 

ttagtggatt tttctttagg aacatgacct ttgggtgagt ataattgaga aattatttta  28312 

attcagaaag tacttttcat tctgttctaa aaatatgtga attgtcttaa gtggtagaaa  28372 

tttgtttctt caaaataaaa ggctcttctc tagatgtttg ggagagctgt atctccaaat  28432 

gacctagtac atcagaaggt cagaccatcc cagcagaaac acacagctgt ttgggtcaca  28492 

gttctgaggg ctgtctttat tccagcgact tcactagctc tgctgactgg ggactgaggt  28552 

gtggttttgt atcccaggac catgttttca acactgaaag gcaaaccaag agtgcatgca  28612 

cttttagaat atgaaacgtg acctgaaata atcccccaag taaatagtgg acaaaaagat  28672 

gagtcaccag ttatcataaa atctcgtttt attgtcacct ccagggtgct tccccccatg  28732 

atgttgcttc taaatgaaag cacagtttgt agacttgaat tgtcacttgc cgataaagaa  28792 

tagattgggc acaaagtaga caacagtatg ggaaaggggc cggaacaatt ggaacaattc  28852 

gcagtaatag agtgagcaga tcagacagca gcagtcagct gttggcgcac actgcaaatg  28912 

aacgctgcct gggttaaatg cttatgctag tttagttttt ttttttttaa gataggatct  28972 

caagtgtcca gggctagcct ttagctctga gcctagtatg gccttgaaca ttgtcttcct  29032 

gccttcaccc gagtactggg attacaggta cgtattccat gcccaggatg gaacccagga  29092 

tttcatgcac cccgggcaga cattgatagc tacatctacc tgactctgct atgttaagga  29152 

taaccattcc agtacctggg ggacaagata ccagaaccac taacaaactg agtttaatca  29212 

aggagttagg agaaagaggc acttttagtc tcaaggaaga aaatcatggg ttgtcagagc  29272 

aggggaaata caggtccagg agaaaaaggc tggccaacag atggcccatg gatgtaggac  29332 

cacacagact gttttaggcc tcactaaggg aggtgtgtag ctcaccttcc tgggggaagg  29392 

catccacaaa cctgtcatct cacaatgaca aaacgtggca ctggcaagaa aactccatgg  29452 

atcaaggtgc cttccatcaa gcattgggac ccacatatcg gaagtagaga acaaaccaac  29512 

ttcacaagtt gtcctctgac tcccacatgc acactgtggc atgcagccac acacacataa  29572 

ataaatgaac agcttttcgt atcaaaatgt ttgccgaaag ctatccagta accagcttat  29632 

tattccgtgc cgcaaagggc agcaccagag tgacgtgctg acggaggccc ctgagctgac  29692 

tgctaatttg ggcctcggcc tcaaaggtgt ccctgagacg gttctgacct gagacactga  29752 

caacatcgga ggggatgggg gcgtgtgtaa acatgagcat gggaaggacc ctcgctgcac  29812 

acagggacat ggcaagccaa gttgggtttt cgaggagggc tgtgtgaaga tgactaggag  29872 

agcttccagc tctcgaatag ctttttacag ggtagataac taagaccaca gactcgggtc  29932 

tgatgggcac agcactgttc tgtggcagag ttttcactag gaagcactct cgtcagatga  29992 

gtgggatgga aggctacctc gttaatcctg agcctgaggg ccaggaatcc aaacagtatc  30052 

tctaggtgtc cactcatcct tccgtgtgcc taccctagac cgatggccat tgcagggagg  30112 

aaggaccgga gggatcaaaa ctgcaacaac aaaaacccga caaaaatgtc aagtggctgg  30172 

ccgccttcat atcgctgctt ggtgatgaga gctgtgtcag atggcctgac cttgtttaca  30232 

gcaagaagac aacacattca ccaacaacac tacagaccac agggtcaccc agtgcctaaa  30292 

ggggcagtgg tgcaatac                                                30310 

 
           
             746  
             20  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            746 

tcgaagacga gctgcagaca                                                 20 

 
           
             747  
             23  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            747 

tggctctatt cttccttggt tga                                             23 

 
           
             748  
             19  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            748 

cagccaggcc agccgctcc                                                  19 

 
           
             749  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            749 

cgttgctgac ctcagagtcc                                                 20 

 
           
             750  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            750 

ctttcagaat ctggctctat                                                 20 

 
           
             751  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            751 

ggcccggcgc tctactccac                                                 20 

 
           
             752  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            752 

gctaaggcaa aggtttgcgg                                                 20 

 
           
             753  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            753 

cgggtccacc aggaggcctg                                                 20 

 
           
             754  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            754 

gccatggcac caggcagtag                                                 20 

 
           
             755  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            755 

gccaggcagc gtgcccagaa                                                 20 

 
           
             756  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            756 

cttccccatt catacaccta                                                 20 

 
           
             757  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            757 

cacttgacac caacagagac                                                 20 

 
           
             758  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            758 

gaagcctgta atcctggcac                                                 20 

 
           
             759  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            759 

gaccatgtcc tggccagaaa                                                 20 

 
           
             760  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            760 

gtcagtccag taagggcttt                                                 20 

 
           
             761  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            761 

ttagcttagc cacagaggga                                                 20 

 
           
             762  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            762 

cgcctgtgct ctcttcctgc                                                 20 

 
           
             763  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            763 

cccatcttct ggcctccttg                                                 20 

 
           
             764  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            764 

ctgaaactcc aggctcagga                                                 20 

 
           
             765  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            765 

ctcatggcag ctgcagcagt                                                 20 

 
           
             766  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            766 

cttgaaaagg aacaaagtgg                                                 20 

 
           
             767  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            767 

tctatacact actcataacc                                                 20 

 
           
             768  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            768 

ccatcacaga ggccacttct                                                 20 

 
           
             769  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            769 

tccatccctg gaacaatgtg                                                 20 

 
           
             770  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            770 

cagagctcag ctttcttccc                                                 20 

 
           
             771  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            771 

agctcacaga gtccagggaa                                                 20 

 
           
             772  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            772 

caagcactgc cagctcacag                                                 20 

 
           
             773  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            773 

tcagagtcca tggcacaagc                                                 20 

 
           
             774  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            774 

ttgccaaaca gaagacacca                                                 20 

 
           
             775  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            775 

gcagagaaac aggctgtggt                                                 20 

 
           
             776  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            776 

gtctgtgatg tgcttggccc                                                 20 

 
           
             777  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            777 

tggagaaagc cgaacaccag                                                 20 

 
           
             778  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            778 

acaggcagtt cccgacccag                                                 20 

 
           
             779  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            779 

ggtctgcctc ccagtaagct                                                 20 

 
           
             780  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            780 

cgtctgtctg cagctcgtct                                                 20 

 
           
             781  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            781 

cttttctgaa tgacttgata                                                 20 

 
           
             782  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            782 

cactgatagg aagtgtgtcc                                                 20 

 
           
             783  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            783 

ctcagttgct gtaaacacag                                                 20 

 
           
             784  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            784 

ccacagcgct ctgagcactc                                                20 

 
           
             785  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            785 

gtcctgaagt atcctgacct                                                 20 

 
           
             786  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            786 

gaaataaact agccagaggg                                                 20 

 
           
             787  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            787 

tttcttcctg actttcagaa                                                 20 

 
           
             788  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            788 

ttgggcgaga tgtctggcaa                                                 20 

 
           
             789  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            789 

cgcctatttg ggcgagatgt                                                 20 

 
           
             790  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            790 

gaactgtgcg gctagctgtc                                                 20 

 
           
             791  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            791 

cgccacaaga gaagactgag                                                 20 

 
           
             792  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            792 

aatgtgtgtg tctttgacag                                                 20 

 
           
             793  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            793 

ctacatgtta tcttcccttc                                                 20 

 
           
             794  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            794 

agggctttgg ccaggcagtt                                                 20 

 
           
             795  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            795 

acagcattgt cattatcagc                                                 20 

 
           
             796  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            796 

gagcaaagat ggtgcgtgac                                                 20 

 
           
             797  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            797 

tgtggaagac atcacggagc                                                 20 

 
           
             798  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            798 

gacagtcgtg tggaagacat                                                 20 

 
           
             799  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            799 

aggttctggt taataaagtt                                                 20 

 
           
             800  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            800 

gtcattttcc agcagtctca                                                 20 

 
           
             801  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            801 

gcgggctcct cagtccatct                                                 20 

 
           
             802  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            802 

gttctctggg acctgtcttc                                                 20 

 
           
             803  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            803 

tcattcccaa gtgggaaccc                                                 20 

 
           
             804  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            804 

cagaagccca cctacatggt                                                 20 

 
           
             805  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            805 

atgcacctct cctaatgctg                                                 20 

 
           
             806  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            806 

gccgatgcac ctctcctaat                                                 20 

 
           
             807  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            807 

gagcacttca gtgtccacta                                                 20 

 
           
             808  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            808 

agatcagcca ttcggctttt                                                 20 

 
           
             809  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            809 

cccatggttt gagatcagcc                                                 20 

 
           
             810  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            810 

gatagaaatc ttgagataat                                                 20 

 
           
             811  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            811 

caccacacag ataagtcgtg                                                 20 

 
           
             812  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            812 

gtaactgaca ccacacagat                                                 20 

 
           
             813  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            813 

agcctgagtg taactgacac                                                 20 

 
           
             814  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            814 

gtagcaagag cctgagtgta                                                 20 

 
           
             815  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            815 

ttgcattccg tagcaagagc                                                 20 

 
           
             816  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            816 

agtgacctgc tgctgtttta                                                 20 

 
           
             817  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            817 

cttttgatat ggaatcttct                                                 20 

 
           
             818  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            818 

aatacagaag cggagggaac                                                 20 

 
           
             819  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            819 

gaggccttgt ctctgaaata                                                 20 

 
           
             820  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            820 

cgtaacaacg cttgaggata                                                 20 

 
           
             821  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            821 

gctgacgatc ccagctttaa                                                 20 

 
           
             822  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            822 

cttgcaggct gtggcggctc                                                 20 

 
           
             823  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            823 

atacttgcag gctgtggcgg                                                 20 

 
           
             824  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            824 

ctgggatgag ttcagaacta                                                 20 

 
           
             825  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            825 

cacatatttt tagaacagaa                                                 20 

 
           
             826  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            826 

gagcctttta ttttgaagaa                                                 20 

 
           
             827  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            827 

ctacgctttc cacgcacagt                                                 20 

 
           
             828  
             3160  
             DNA  
             Homo sapiens  
             
               CDS  
               (1035)...(2246)  
             
           
            828 

cctcccctcg cccggcgcgg tcccgtccgc ctctcgctcg cctcccgcct cccctcggtc     60 

ttccgaggcg cccgggctcc cggcgcggcg gcggaggggg cgggcaggcc ggcgggcggt    120 

gatgtggcag gactctttat gcgctgcggc aggatacgcg ctcggcgctg ggacgcgact    180 

gcgctcagtt ctctcctctc ggaagctgca gccatgatgg aagtttgaga gttgagccgc    240 

tgtgaggcga ggccgggctc aggcgaggga gatgagagac ggcggcggcc gcggcccgga    300 

gcccctctca gcgcctgtga gcagccgcgg gggcagcgcc ctcggggagc cggccggcct    360 

gcggcggcgg cagcggcggc gtttctcgcc tcctcttcgt cttttctaac cgtgcagcct    420 

cttcctcggc ttctcctgaa agggaaggtg gaagccgtgg gctcgggcgg gagccggctg    480 

aggcgcggcg gcggcggcgg cggcacctcc cgctcctgga gcggggggga gaagcggcgg    540 

cggcggcggc cgcggcggct gcagctccag ggagggggtc tgagtcgcct gtcaccattt    600 

ccagggctgg gaacgccgga gagttggtct ctccccttct actgcctcca acacggcggc    660 

ggcggcggcg gcacatccag ggacccgggc cggttttaaa cctcccgtcc gccgccgccg    720 

caccccccgt ggcccgggct ccggaggccg ccggcggagg cagccgttcg gaggattatt    780 

cgtcttctcc ccattccgct gccgccgctg ccaggcctct ggctgctgag gagaagcagg    840 

cccagtcgct gcaaccatcc agcagccgcc gcagcagcca ttacccggct gcggtccaga    900 

gccaagcggc ggcagagcga ggggcatcag ctaccgccaa gtccagagcc atttccatcc    960 

tgcagaagaa gccccgccac cagcagcttc tgccatctct ctcctccttt ttcttcagcc   1020 

acaggctccc agac atg aca gcc atc atc aaa gag atc gtt agc aga aac     1070 
                Met Thr Ala Ile Ile Lys Glu Ile Val Ser Arg Asn 
                 1               5                   10 

aa agg aga tat caa gag gat gga ttc gac tta gac ttg acc tat att      1118 
Lys Arg Arg Tyr Gln Glu Asp Gly Phe Asp Leu Asp Leu Thr Tyr Ile 
         15                  20                  25 

at cca aac att att gct atg gga ttt cct gca gaa aga ctt gaa ggc      1166 
Tyr Pro Asn Ile Ile Ala Met Gly Phe Pro Ala Glu Arg Leu Glu Gly 
     30                  35                  40 

ta tac agg aac aat att gat gat gta gta agg ttt ttg gat tca aag      1214 
Val Tyr Arg Asn Asn Ile Asp Asp Val Val Arg Phe Leu Asp Ser Lys 
 45                  50                  55                  60 

at aaa aac cat tac aag ata tac aat ctt tgt gct gaa aga cat tat      1262 
His Lys Asn His Tyr Lys Ile Tyr Asn Leu Cys Ala Glu Arg His Tyr 
                 65                  70                  75 

ac acc gcc aaa ttt aat tgc aga gtt gca caa tat cct ttt gaa gac      1310 
Asp Thr Ala Lys Phe Asn Cys Arg Val Ala Gln Tyr Pro Phe Glu Asp 
             80                  85                  90 

at aac cca cca cag cta gaa ctt atc aaa ccc ttt tgt gaa gat ctt      1358 
His Asn Pro Pro Gln Leu Glu Leu Ile Lys Pro Phe Cys Glu Asp Leu 
         95                 100                 105 

ac caa tgg cta agt gaa gat gac aat cat gtt gca gca att cac tgt      1406 
Asp Gln Trp Leu Ser Glu Asp Asp Asn His Val Ala Ala Ile His Cys 
    110                 115                 120 

aa gct gga aag gga cga act ggt gta atg ata tgt gca tat tta tta      1454 
Lys Ala Gly Lys Gly Arg Thr Gly Val Met Ile Cys Ala Tyr Leu Leu 
125                 130                 135                 140 

at cgg ggc aaa ttt tta aag gca caa gag gcc cta gat ttc tat ggg      1502 
His Arg Gly Lys Phe Leu Lys Ala Gln Glu Ala Leu Asp Phe Tyr Gly 
                145                 150                 155 

aa gta agg acc aga gac aaa aag gga gta act att ccc agt cag agg      1550 
Glu Val Arg Thr Arg Asp Lys Lys Gly Val Thr Ile Pro Ser Gln Arg 
            160                 165                 170 

gc tat gtg tat tat tat agc tac ctg tta aag aat cat ctg gat tat      1598 
Arg Tyr Val Tyr Tyr Tyr Ser Tyr Leu Leu Lys Asn His Leu Asp Tyr 
        175                 180                 185 

ga cca gtg gca ctg ttg ttt cac aag atg atg ttt gaa act att cca      1646 
Arg Pro Val Ala Leu Leu Phe His Lys Met Met Phe Glu Thr Ile Pro 
    190                 195                 200 

tg ttc agt ggc gga act tgc aat cct cag ttt gtg gtc tgc cag cta      1694 
Met Phe Ser Gly Gly Thr Cys Asn Pro Gln Phe Val Val Cys Gln Leu 
205                 210                 215                 220 

ag gtg aag ata tat tcc tcc aat tca gga ccc aca cga cgg gaa gac      1742 
Lys Val Lys Ile Tyr Ser Ser Asn Ser Gly Pro Thr Arg Arg Glu Asp 
                225                 230                 235 

ag ttc atg tac ttt gag ttc cct cag ccg tta cct gtg tgt ggt gat      1790 
Lys Phe Met Tyr Phe Glu Phe Pro Gln Pro Leu Pro Val Cys Gly Asp 
            240                 245                 250 

tc aaa gta gag ttc ttc cac aaa cag aac aag atg cta aaa aag gac      1838 
Ile Lys Val Glu Phe Phe His Lys Gln Asn Lys Met Leu Lys Lys Asp 
        255                 260                 265 

aa atg ttt cac ttt tgg gta aat aca ttc ttc ata cca gga cca gag      1886 
Lys Met Phe His Phe Trp Val Asn Thr Phe Phe Ile Pro Gly Pro Glu 
    270                 275                 280 

aa acc tca gaa aaa gta gaa aat gga agt cta tgt gat caa gaa atc      1934 
Glu Thr Ser Glu Lys Val Glu Asn Gly Ser Leu Cys Asp Gln Glu Ile 
285                 290                 295                 300 

at agc att tgc agt ata gag cgt gca gat aat gac aag gaa tat cta      1982 
Asp Ser Ile Cys Ser Ile Glu Arg Ala Asp Asn Asp Lys Glu Tyr Leu 
                305                 310                 315 

ta ctt act tta aca aaa aat gat ctt gac aaa gca aat aaa gac aaa      2030 
Val Leu Thr Leu Thr Lys Asn Asp Leu Asp Lys Ala Asn Lys Asp Lys 
            320                 325                 330 

cc aac cga tac ttt tct cca aat ttt aag gtg aag ctg tac ttc aca      2078 
Ala Asn Arg Tyr Phe Ser Pro Asn Phe Lys Val Lys Leu Tyr Phe Thr 
        335                 340                 345 

aa aca gta gag gag ccg tca aat cca gag gct agc agt tca act tct      2126 
Lys Thr Val Glu Glu Pro Ser Asn Pro Glu Ala Ser Ser Ser Thr Ser 
    350                 355                 360 

ta aca cca gat gtt agt gac aat gaa cct gat cat tat aga tat tct      2174 
Val Thr Pro Asp Val Ser Asp Asn Glu Pro Asp His Tyr Arg Tyr Ser 
365                 370                 375                 380 

ac acc act gac tct gat cca gag aat gaa cct ttt gat gaa gat cag      2222 
Asp Thr Thr Asp Ser Asp Pro Glu Asn Glu Pro Phe Asp Glu Asp Gln 
                385                 390                 395 

at aca caa att aca aaa gtc tga attttttttt atcaagaggg ataaaacacc     2276 
His Thr Gln Ile Thr Lys Val  * 
            400 

atgaaaataa acttgaataa actgaaaatg gacctttttt tttttaatgg caataggaca   2336 

ttgtgtcaga ttaccagtta taggaacaat tctcttttcc tgaccaatct tgttttaccc   2396 

tatacatcca cagggttttg acacttgttg tccagttgaa aaaaggttgt gtagctgtgt   2456 

catgtatata cctttttgtg tcaaaaggac atttaaaatt caattaggat taataaagat   2516 

ggcactttcc cgttttattc cagttttata aaaagtggag acagactgat gtgtatacgt   2576 

aggaattttt tccttttgtg ttctgtcacc aactgaagtg gctaaagagc tttgtgatat   2636 

actggttcac atcctacccc tttgcacttg tggcaacaga taagtttgca gttggctaag   2696 

agaggtttcc gaaaggtttt gctaccattc taatgcatgt attcgggtta gggcaatgga   2756 

ggggaatgct cagaaaggaa ataattttat gctggactct ggaccatata ccatctccag   2816 

ctatttacac acacctttct ttagcatgct acagttatta atctggacat tcgaggaatt   2876 

ggccgctgtc actgcttgtt gtttgcgcat ttttttttaa agcatattgg tgctagaaaa   2936 

ggcagctaaa ggaagtgaat ctgtattggg gtacaggaat gaaccttctg caacatctta   2996 

agatccacaa atgaagggat ataaaaataa tgtcataggt aagaaacaca gcaacaatga   3056 

cttaaccata taaatgtgga ggctatcaac aaagaatggg cttgaaacat tataaaaatt   3116 

gacaatgatt tattaaatat gttttctcaa ttgtaaaaaa aaaa                    3160 

 
           
             829  
             26  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            829 

aatggctaag tgaagatgac aatcat                                          26 

 
           
             830  
             25  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            830 

tgcacatatc attacaccag ttcgt                                           25 

 
           
             831  
             30  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            831 

ttgcagcaat tcactgtaaa gctggaaagg                                      30 

 
           
             832  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            832 

cgagaggcgg acgggacc                                                   18 

 
           
             833  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            833 

cgggcgcctc ggaagacc                                                   18 

 
           
             834  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            834 

tggctgcagc ttccgaga                                                   18 

 
           
             835  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            835 

cccgcggctg ctcacagg                                                   18 

 
           
             836  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            836 

caggagaagc cgaggaag                                                   18 

 
           
             837  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            837 

gggaggtgcc gccgccgc                                                   18 

 
           
             838  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            838 

ccgggtccct ggatgtgc                                                   18 

 
           
             839  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            839 

cctccgaacg gctgcctc                                                   18 

 
           
             840  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            840 

tctcctcagc agccagag                                                   18 

 
           
             841  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            841 

cgcttggctc tggaccgc                                                   18 

 
           
             842  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            842 

tcttctgcag gatggaaa                                                   18 

 
           
             843  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            843 

ggataaatat aggtcaag                                                   18 

 
           
             844  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            844 

tcaatattgt tcctgtat                                                   18 

 
           
             845  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            845 

ttaaatttgg cggtgtca                                                   18 

 
           
             846  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            846 

caagatcttc acaaaagg                                                   18 

 
           
             847  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            847 

attacaccag ttcgtccc                                                   18 

 
           
             848  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            848 

tgtctctggt ccttactt                                                   18 

 
           
             849  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            849 

acatagcgcc tctgactg                                                   18 

 
           
             850  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            850 

gaatatatct tcaccttt                                                   18 

 
           
             851  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            851 

ggaagaactc tactttga                                                   18 

 
           
             852  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            852 

tgaagaatgt atttaccc                                                   18 

 
           
             853  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            853 

ggttggcttt gtctttat                                                   18 

 
           
             854  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            854 

tgctagcctc tggatttg                                                   18 

 
           
             855  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            855 

tctggatcag agtcagtg                                                   18 

 
           
             856  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            856 

tattttcatg gtgtttta                                                   18 

 
           
             857  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            857 

tgttcctata actggtaa                                                   18 

 
           
             858  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            858 

gtgtcaaaac cctgtgga                                                   18 

 
           
             859  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            859 

actggaataa aacgggaa                                                   18 

 
           
             860  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            860 

acttcagttg gtgacaga                                                   18 

 
           
             861  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            861 

tagcaaaacc tttcggaa                                                   18 

 
           
             862  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            862 

aattatttcc tttctgag                                                   18 

 
           
             863  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            863 

gctg gagatggt                                                         18 

 
           
             864  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            864 

cagattaata actgtagc                                                   18 

 
           
             865  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            865 

ccccaataca gattcact                                                   18 

 
           
             866  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            866 

attgttgctg tgtttctt                                                   18 

 
           
             867  
             18  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            867 

tgtttcaagc ccattctt                                                   18 

 
           
             868  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            868 

ctgctagcct ctggatttga                                                 20 

 
           
             869  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            869 

acatagcgcc tctgactggg                                                 20 

 
           
             870  
             20  
             DNA  
             Artificial Sequence  
             
               Antisense Oligonucleotide  
             
           
            870 

cttctggcat ccggtttaga                                                 20 

 
           
             871  
             2160  
             DNA  
             Mus musculus  
             
               CDS  
               (949)...(2160)  
             
           
            871 

ggcgccctgc tctcccggcg gggcggcgga gggggcgggc tggccggcgc acggtgatgt     60 

ggcgggactc tttgtgcact gcggcaggat acgcgcttgg gcgtcgggac gcggctgcgc    120 

tcagctctct cctctcggaa gctgcagcca tgatggaagt ttgagagttg agccgctgtg    180 

aggccaggcc cggcgcaggc gagggagatg agagacggcg gcggccacgg cccagagccc    240 

ctctcagcgc ctgtgagcag ccgcgggggc agcgccctcg gggagccggc cgggcggcgg    300 

cggcggcagc ggcggcgggc ctcgcctcct cgtcgtctgt tctaaccggg cagcttctga    360 

gcagcttcgg agagagacgg tggaagaagc cgtgggctcg agcgggagcc ggcgcaggct    420 

cggcggctgc acctcccgct cctggagcgg gggggagaag cggcggcggc ggccgcggct    480 

ccggggaggg ggtcggagtc gcctgtcacc attgccaggg ctgggaacgc cggagagttg    540 

ctctctcccc ttctcctgcc tccaacacgg cggcggcggc ggcggcacgt ccagggaccc    600 

gggccggtgt taagcctccc gtccgccgcc gccgcacccc ccctggcccg ggctccggag    660 

gccgccggag gaggcagccg ctgcgaggat tatccgtctt ctccccattc cgctgcctcg    720 

gctgccaggc ctctggctgc tgaggagaag caggcccagt ctctgcaacc atccagcagc    780 

cgccgcagca gccattaccc ggctgcggtc cagggccaag cggcagcaga gcgaggggca    840 

tcagcgaccg ccaagtccag agccatttcc atcctgcaga agaagcctcg ccaccagcag    900 

cttctgccat ctctctcctc ctttttcttc agccacaggc tcccagac atg aca gcc     957 
                                                     Met Thr Ala 
                                                      1 

atc atc aaa gag atc gtt agc aga aac aaa agg aga tat caa gag gat     1005 
Ile Ile Lys Glu Ile Val Ser Arg Asn Lys Arg Arg Tyr Gln Glu Asp 
     5                   10                  15 

gga ttc gac tta gac ttg acc tat att tat cca aat att att gct atg     1053 
Gly Phe Asp Leu Asp Leu Thr Tyr Ile Tyr Pro Asn Ile Ile Ala Met 
 20                  25                  30                  35 

gga ttt cct gca gaa aga ctt gaa ggt gta tac agg aac aat att gat     1101 
Gly Phe Pro Ala Glu Arg Leu Glu Gly Val Tyr Arg Asn Asn Ile Asp 
                 40                  45                  50 

gat gta gta agg ttt ttg gat tca aag cat aaa aac cat tac aag ata     1149 
Asp Val Val Arg Phe Leu Asp Ser Lys His Lys Asn His Tyr Lys Ile 
             55                  60                  65 

tac aat cta tgt gct gag aga cat tat gac acc gcc aaa ttt aac tgc     1197 
Tyr Asn Leu Cys Ala Glu Arg His Tyr Asp Thr Ala Lys Phe Asn Cys 
         70                  75                  80 

aga gtt gca cag tat cct ttt gaa gac cat aac cca cca cag cta gaa     1245 
Arg Val Ala Gln Tyr Pro Phe Glu Asp His Asn Pro Pro Gln Leu Glu 
     85                  90                  95 

ctt atc aaa ccc ttc tgt gaa gat ctt gac caa tgg cta agt gaa gat     1293 
Leu Ile Lys Pro Phe Cys Glu Asp Leu Asp Gln Trp Leu Ser Glu Asp 
100                 105                 110                 115 

gac aat cat gtt gca gca att cac tgt aaa gct gga aag gga cgg act     1341 
Asp Asn His Val Ala Ala Ile His Cys Lys Ala Gly Lys Gly Arg Thr 
                120                 125                 130 

ggt gta atg att tgt gca tat tta ttg cat cgg ggc aaa ttt tta aag     1389 
Gly Val Met Ile Cys Ala Tyr Leu Leu His Arg Gly Lys Phe Leu Lys 
            135                 140                 145 

gca caa gag gcc cta gat ttt tat ggg gaa gta agg acc aga gac aaa     1437 
Ala Gln Glu Ala Leu Asp Phe Tyr Gly Glu Val Arg Thr Arg Asp Lys 
        150                 155                 160 

aag gga gtc aca att ccc agt cag agg cgc tat gta tat tat tat agc     1485 
Lys Gly Val Thr Ile Pro Ser Gln Arg Arg Tyr Val Tyr Tyr Tyr Ser 
    165                 170                 175 

tac ctg cta aaa aat cac ctg gat tac aga ccc gtg gca ctg ctg ttt     1533 
Tyr Leu Leu Lys Asn His Leu Asp Tyr Arg Pro Val Ala Leu Leu Phe 
180                 185                 190                 195 

cac aag atg atg ttt gaa act att cca atg ttc agt ggc gga act tgc     1581 
His Lys Met Met Phe Glu Thr Ile Pro Met Phe Ser Gly Gly Thr Cys 
                200                 205                 210 

aat cct cag ttt gtg gtc tgc cag cta aag gtg aag ata tat tcc tcc     1629 
Asn Pro Gln Phe Val Val Cys Gln Leu Lys Val Lys Ile Tyr Ser Ser 
            215                 220                 225 

aat tca gga ccc acg cgg cgg gag gac aag ttc atg tac ttt gag ttc     1677 
Asn Ser Gly Pro Thr Arg Arg Glu Asp Lys Phe Met Tyr Phe Glu Phe 
        230                 235                 240 

cct cag cca ttg cct gtg tgt ggt gat atc aaa gta gag ttc ttc cac     1725 
Pro Gln Pro Leu Pro Val Cys Gly Asp Ile Lys Val Glu Phe Phe His 
    245                 250                 255 

aaa cag aac aag atg ctc aaa aag gac aaa atg ttt cac ttt tgg gta     1773 
Lys Gln Asn Lys Met Leu Lys Lys Asp Lys Met Phe His Phe Trp Val 
260                 265                 270                 275 

aat acg ttc ttc ata cca gga cca gag gaa acc tca gaa aaa gtg gaa     1821 
Asn Thr Phe Phe Ile Pro Gly Pro Glu Glu Thr Ser Glu Lys Val Glu 
                280                 285                 290 

aat gga agt ctt tgt gat cag gaa atc gat agc att tgc agt ata gag     1869 
Asn Gly Ser Leu Cys Asp Gln Glu Ile Asp Ser Ile Cys Ser Ile Glu 
            295                 300                 305 

cgt gca gat aat gac aag gag tat ctt gta ctc acc cta aca aaa aac     1917 
Arg Ala Asp Asn Asp Lys Glu Tyr Leu Val Leu Thr Leu Thr Lys Asn 
        310                 315                 320 

gat ctt gac aaa gca aac aaa gac aag gcc aac cga tac ttc tct cca     1965 
Asp Leu Asp Lys Ala Asn Lys Asp Lys Ala Asn Arg Tyr Phe Ser Pro 
    325                 330                 335 

aat ttt aag gtg aaa cta tac ttt aca aaa aca gta gag gag cca tca     2013 
Asn Phe Lys Val Lys Leu Tyr Phe Thr Lys Thr Val Glu Glu Pro Ser 
340                 345                 350                 355 

aat cca gag gct agc agt tca act tct gtg act cca gat gtt agt gac     2061 
Asn Pro Glu Ala Ser Ser Ser Thr Ser Val Thr Pro Asp Val Ser Asp 
                360                 365                 370 

aat gaa cct gat cat tat aga tat tct gac acc act gac tct gat cca     2109 
Asn Glu Pro Asp His Tyr Arg Tyr Ser Asp Thr Thr Asp Ser Asp Pro 
            375                 380                 385 

gag aat gaa cct ttt gat gaa gat cag cat tca caa att aca aaa gtc     2157 
Glu Asn Glu Pro Phe Asp Glu Asp Gln His Ser Gln Ile Thr Lys Val 
        390                 395                 400  
           
             872  
             24  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            872 

atgacaatca tgttgcagca attc                                            24 

 
           
             873  
             25  
             DNA  
             Artificial Sequence  
             
               PCR Primer  
             
           
            873 

cgatgcaata aatatgcaca aatca                                           25 

 
           
             874  
             28  
             DNA  
             Artificial Sequence  
             
               PCR Probe  
             
           
            874 

ctgtaaagct ggaaagggac ggactggt                                        28 

 
           
             875  
             1212  
             DNA  
             Rattus norvegicus  
             
               CDS  
               (1)...(1212)  
             
           
            875 

atg aca gcc atc atc aaa gag atc gtt agc aga aac aaa agg aga tat       48 
Met Thr Ala Ile Ile Lys Glu Ile Val Ser Arg Asn Lys Arg Arg Tyr 
 1               5                   10                  15 

caa gag gat gga ttc gac tta gac ttg acc tat att tat cca aat att       96 
Gln Glu Asp Gly Phe Asp Leu Asp Leu Thr Tyr Ile Tyr Pro Asn Ile 
             20                  25                  30 

att gct atg gga ttt cct gca gaa aga ctt gaa ggt gta tac agg aac      144 
Ile Ala Met Gly Phe Pro Ala Glu Arg Leu Glu Gly Val Tyr Arg Asn 
         35                  40                  45 

aat att gat gat gta gta agg ttt ttg gat tca aag cat aaa aac cat      192 
Asn Ile Asp Asp Val Val Arg Phe Leu Asp Ser Lys His Lys Asn His 
     50                  55                  60 

tac aag ata tac aat cta tgt gct gag aga cat tat gac acc gcc aaa      240 
Tyr Lys Ile Tyr Asn Leu Cys Ala Glu Arg His Tyr Asp Thr Ala Lys 
 65                  70                  75                  80 

ttt aac tgc aga gtt gca cag tat cct ttt gaa gac cat aac cca cca      288 
Phe Asn Cys Arg Val Ala Gln Tyr Pro Phe Glu Asp His Asn Pro Pro 
                 85                  90                  95 

cag cta gaa ctt atc aaa ccc ttt tgt gaa gat ctt gac caa tgg cta      336 
Gln Leu Glu Leu Ile Lys Pro Phe Cys Glu Asp Leu Asp Gln Trp Leu 
            100                 105                 110 

agt gaa gac gac aat cat gtt gca gca att cac tgt aaa gct ggg aaa      384 
Ser Glu Asp Asp Asn His Val Ala Ala Ile His Cys Lys Ala Gly Lys 
        115                 120                 125 

gga cgg act ggt gta atg att tgt gca tat tta ttg cat cgg ggc aag      432 
Gly Arg Thr Gly Val Met Ile Cys Ala Tyr Leu Leu His Arg Gly Lys 
    130                 135                 140 

ttt tta aag gca caa gag gcc ctg gat ttt tat ggg gaa gta agg acc      480 
Phe Leu Lys Ala Gln Glu Ala Leu Asp Phe Tyr Gly Glu Val Arg Thr 
145                 150                 155                 160 

aga gat aaa aag gga gta act att ccc agt cag agg cgc tat gta tat      528 
Arg Asp Lys Lys Gly Val Thr Ile Pro Ser Gln Arg Arg Tyr Val Tyr 
                165                 170                 175 

tat tat agc tac ctg tta aag aat cac ctg gat tac aga cca gtg gca      576 
Tyr Tyr Ser Tyr Leu Leu Lys Asn His Leu Asp Tyr Arg Pro Val Ala 
            180                 185                 190 

ctg ttg ttt cac aag atg atg ttt gaa act att cca atg ttc agt ggc      624 
Leu Leu Phe His Lys Met Met Phe Glu Thr Ile Pro Met Phe Ser Gly 
        195                 200                 205 

gga act tgc aat ccc cag ttt gtg gtc tgc cag cta aag gtg aag atc      672 
Gly Thr Cys Asn Pro Gln Phe Val Val Cys Gln Leu Lys Val Lys Ile 
    210                 215                 220 

tac tcc tcc aac tca gga ccc acg cgg cgg gag gac aag ctc atg tac      720 
Tyr Ser Ser Asn Ser Gly Pro Thr Arg Arg Glu Asp Lys Leu Met Tyr 
225                 230                 235                 240 

ttt gag ttc cct cag cca ttg cct gtg tgt ggt gac atc aaa gta gag      768 
Phe Glu Phe Pro Gln Pro Leu Pro Val Cys Gly Asp Ile Lys Val Glu 
                245                 250                 255 

ttc ttc cac aaa cag aac aag atg ctc aaa aag gac aaa atg ttt cac      816 
Phe Phe His Lys Gln Asn Lys Met Leu Lys Lys Asp Lys Met Phe His 
            260                 265                 270 

ttt tgg gta aat acg ttc ttc ata cca gga cca gag gaa acc tca gaa      864 
Phe Trp Val Asn Thr Phe Phe Ile Pro Gly Pro Glu Glu Thr Ser Glu 
        275                 280                 285 

aaa gtg gaa aat gga agt ctt tgt gat cag gaa atc gat agc att tgt      912 
Lys Val Glu Asn Gly Ser Leu Cys Asp Gln Glu Ile Asp Ser Ile Cys 
    290                 295                 300 

agt ata gag cgt gcg gat aat gac aag gag tat ctt gtg ctc acc ctg      960 
Ser Ile Glu Arg Ala Asp Asn Asp Lys Glu Tyr Leu Val Leu Thr Leu 
305                 310                 315                 320 

aca aaa aat gat ctt gac aaa gca aac aaa gac aag gcc aac cga tac     1008 
Thr Lys Asn Asp Leu Asp Lys Ala Asn Lys Asp Lys Ala Asn Arg Tyr 
                325                 330                 335 

ttc tct cca aat ttt aag gtg aag tta tac ttc aca aaa aca gta gag     1056 
Phe Ser Pro Asn Phe Lys Val Lys Leu Tyr Phe Thr Lys Thr Val Glu 
            340                 345                 350 

gag cca tca aat cca gag gct agc agt tca act tct gtg act cca gac     1104 
Glu Pro Ser Asn Pro Glu Ala Ser Ser Ser Thr Ser Val Thr Pro Asp 
        355                 360                 365 

gtt agt gac aat gaa cct gat cat tat aga tat tct gac acc act gac     1152 
Val Ser Asp Asn Glu Pro Asp His Tyr Arg Tyr Ser Asp Thr Thr Asp 
    370                 375                 380 

tct gat cca gag aat gaa cct ttt gat gaa gat cag cat tca caa att     1200 
Ser Asp Pro Glu Asn Glu Pro Phe Asp Glu Asp Gln His Ser Gln Ile 
385                 390                 395                 400 

aca aaa gtc tga                                                     1212 
Thr Lys Val  * 

 
           
             876  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            876 

cgagaggcgg acgggaccgt t                                               21 

 
           
             877  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            877 

cgggcgcctc ggaagaccgt t                                               21 

 
           
             878  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            878 

tggctgcagc ttccgagagt t                                               21 

 
           
             879  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            879 

cccgcggctg ctcacaggct t                                               21 

 
           
             880  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            880 

caggagaagc cgaggaagat t                                               21 

 
           
             881  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            881 

gggaggtgcc gccgccgcct t                                               21 

 
           
             882  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            882 

ccgggtccct ggatgtgcct t                                               21 

 
           
             883  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            883 

cctccgaacg gctgcctcct t                                               21 

 
           
             884  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            884 

tctcctcagc agccagaggt t                                               21 

 
           
             885  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            885 

cgcttggctc tggaccgcat t                                               21 

 
           
             886  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            886 

tcttctgcag gatggaaatt t                                               21 

 
           
             887  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            887 

ggataaatat aggtcaagtt t                                               21 

 
           
             888  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            888 

tcaatattgt tcctgtatat t                                               21 

 
           
             889  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            889 

ttaaatttgg cggtgtcatt t                                               21 

 
           
             890  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            890 

caagatcttc acaaaagggt t                                               21 

 
           
             891  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            891 

attacaccag ttcgtccctt t                                               21 

 
           
             892  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            892 

tgtctctggt ccttacttct t                                               21 

 
           
             893  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            893 

acatagcgcc tctgactggt t                                               21 

 
           
             894  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            894 

gaatatatct tcacctttat t                                               21 

 
           
             895  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            895 

ggaagaactc tactttgatt t                                               21 

 
           
             896  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            896 

tgaagaatgt atttacccat t                                               21 

 
           
             897  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            897 

ggttggcttt gtctttattt t                                               21 

 
           
             898  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            898 

tgctagcctc tggatttgat t                                               21 

 
           
             899  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            899 

tctggatcag agtcagtggt t                                               21 

 
           
             900  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            900 

tattttcatg gtgttttact t                                               21 

 
           
             901  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            901 

tgttcctata actggtaatt t                                               21 

 
           
             902  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            902 

gtgtcaaaac cctgtggatt t                                               21 

 
           
             903  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            903 

actggaataa aacgggaaat t                                               21 

 
           
             904  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            904 

acttcagttg gtgacagaat t                                               21 

 
           
             905  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            905 

tagcaaaacc tttcggaaat t                                               21 

 
           
             906  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            906 

aattatttcc tttctgagct t                                               21 

 
           
             907  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            907 

taaatagctg gagatggtct t                                               21 

 
           
             908  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            908 

cagattaata actgtagcat t                                               21 

 
           
             909  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            909 

ccccaataca gattcacttt t                                               21 

 
           
             910  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            910 

attgttgctg tgtttcttat t                                               21 

 
           
             911  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            911 

tgtttcaagc ccattctttt t                                               21 

 
           
             912  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            912 

cggtcccgtc cgcctctcgt t                                               21 

 
           
             913  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            913 

cggtcttccg aggcgcccgt t                                               21 

 
           
             914  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            914 

ctctcggaag ctgcagccat t                                               21 

 
           
             915  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            915 

gcctgtgagc agccgcgggt t                                               21 

 
           
             916  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            916 

tcttcctcgg cttctcctgt t                                               21 

 
           
             917  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            917 

ggcggcggcg gcacctccct t                                               21 

 
           
             918  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            918 

ggcacatcca gggacccggt t                                               21 

 
           
             919  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            919 

ggaggcagcc gttcggaggt t                                               21 

 
           
             920  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            920 

cctctggctg ctgaggagat t                                               21 

 
           
             921  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            921 

tgcggtccag agccaagcgt t                                               21 

 
           
             922  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            922 

atttccatcc tgcagaagat t                                               21 

 
           
             923  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            923 

acttgaccta tatttatcct t                                               21 

 
           
             924  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            924 

tatacaggaa caatattgat t                                               21 

 
           
             925  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            925 

atgacaccgc caaatttaat t                                               21 

 
           
             926  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            926 

cccttttgtg aagatcttgt t                                               21 

 
           
             927  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            927 

agggacgaac tggtgtaatt t                                               21 

 
           
             928  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            928 

gaagtaagga ccagagacat t                                               21 

 
           
             929  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            929 

ccagtcagag gcgctatgtt t                                               21 

 
           
             930  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            930 

taaaggtgaa gatatattct t                                               21 

 
           
             931  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            931 

atcaaagtag agttcttcct t                                               21 

 
           
             932  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            932 

tgggtaaata cattcttcat t                                               21 

 
           
             933  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            933 

aataaagaca aagccaacct t                                               21 

 
           
             934  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            934 

tcaaatccag aggctagcat t                                               21 

 
           
             935  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            935 

ccactgactc tgatccagat t                                               21 

 
           
             936  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            936 

gtaaaacacc atgaaaatat t                                               21 

 
           
             937  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            937 

attaccagtt ataggaacat t                                               21 

 
           
             938  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            938 

atccacaggg ttttgacact t                                               21 

 
           
             939  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            939 

tttcccgttt tattccagtt t                                               21 

 
           
             940  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            940 

ttctgtcacc aactgaagtt t                                               21 

 
           
             941  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            941 

tttccgaaag gttttgctat t                                               21 

 
           
             942  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            942 

gctcagaaag gaaataattt t                                               21 

 
           
             943  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            943 

gaccatctcc agctatttat t                                               21 

 
           
             944  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            944 

tgctacagtt attaatctgt t                                               21 

 
           
             945  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            945 

aagtgaatct gtattggggt t                                               21 

 
           
             946  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            946 

taagaaacac agcaacaatt t                                               21 

 
           
             947  
             21  
             DNA  
             Artificial Sequence  
             
               Oligomeric Compound  
             
           
            947 

aaagaatggg cttgaaacat t                                               21