Patent Publication Number: US-2002009739-A1

Title: Metastatic breast and colon cancer regulated genes

Description:
TECHNICAL FIELD OF THE INVENTION  
       [0001] This invention relates to methods for predicting the behavior of tumors. More particularly, the invention relates to methods in which a tumor sample is examined for expression of a specified gene sequence thereby to indicate propensity for metastatic spread.  
       BACKGROUND OF THE INVENTION  
       [0002] Breast cancer is one of the most common malignant diseases in women, with about 1,000,000 new cases per year worldwide. Colon cancer is another of the most common cancers. Despite use of a number of histochemical, genetic, and immunological markers, clinicians still have a difficult time predicting which tumors will metastasize to other organs. Some patients are in need of adjuvant therapy to prevent recurrence and metastasis and others are not. However, distinguishing between these subpopulations of patients is not straightforward, and course of treatment is not easily charted. There is a need in the art for new markers for distinguish between tumors which will or have metastasized and those which are less likely to metastasize  
       SUMMARY OF THE INVENTION  
       [0003] It is an object of the present invention to provide markers for distinguishing between tumors which will or have metastasized and those which are less likely to metastasize. These and other objects of the invention are provided by one or more of the embodiments described below.  
       [0004] One embodiment of the invention provides an isolated and purified human protein having an amino acid sequence which is at least 85% identical to an amino acid sequence encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOS:1-63 or the complement thereof.  
       [0005] Another embodiment of the invention provides a fusion protein which comprises a first protein segment and a second protein segment fused to each other by means of a peptide bond. The first protein segment consists of at least six contiguous amino acids selected from an amino acid sequence encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOS:1-63 or the complement thereof.  
       [0006] Yet another embodiment of the invention provides an isolated and purified polypeptide consisting of at least six contiguous amino acids of a human protein having an amino acid sequence encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOS: 1-63 or the complement thereof.  
       [0007] Still another embodiment of the invention provides a preparation of antibodies which specifically bind to a human protein which comprises an amino acid sequence encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOS: 1-63 or the complement thereof.  
       [0008] Even another embodiment of the invention provides an isolated and purified subgenomic polynucteotide comprising at least 11 contiguous nucleotides of a nucleotide sequence which is at least 96% identical to a nucleotide sequence selected from the group consisting of SEQ ID NOS: 1-63 or the complement thereof.  
       [0009] Another embodiment of the invention provides an isolated and purified gene which comprises a coding sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS: 1-63 or the complement thereof.  
       [0010] Yet another embodiment of the invention provides a method for determining metastasis in a tissue sample. An expression product of a gene which comprises a coding sequence selected from the group consisting of SEQ ID NOS:1, 2, 4, 5, 9, 11, 13, 14, 18, 19, 20, 22, 24, 26, 29, 30, 33, 35, 36, 38-41, 45, 48, 52, 55, 57, 58, 60, 63-66, 69-74, 76, 80, 82, and 83 is measured in a tissue sample. A tissue sample which expresses the product is categorized as metastatic.  
       [0011] Still another embodiment of the invention provides a method for determining metastasis in a tissue sample. An expression product of a gene which comprises a sequence selected from the group consisting of SEQ ID NOS:3, 7, 8, 10, 12, 15-17, 21, 23, 28, 31, 34, 37, 42-44, 46, 47, 49-51, 53, 59, 61, 62, 67, 68, 75, 77-79, 81, 84, and 85 is measured in a tissue sample. A tissue sample which does not express the product is categorized as metastatic.  
       [0012] Even another embodiment of the invention provides a method for determining metastatic potential in a tissue sample. An expression product of a gene which comprises a sequence selected from the group consisting of SEQ ID NOS:1, 2, 4, 5, 9, 11, 13, 14, 18, 19, 20, 22, 24, 26, 29, 30, 33, 35, 36, 38-41, 45, 48, 52, 55, 57, 58, 60, 63-66, 69-74, 76, 80, 82, and 83 is measured in a tissue sample. A tissue sample which expresses the product is categorized as having metastatic potential.  
       [0013] A further embodiment of the invention provides a method for determining metastatic potential in a tissue sample. An expression product of a gene which comprises a sequence selected from the group consisting of SEQ ID NOS:3, 7, 8, 10, 12, 15-17, 21, 23, 28, 31, 34, 37, 42-44, 46, 47, 49-51, 53, 59, 61, 62, 67, 68, 75, 77-79, 81, 84, and 85 is measured in a tissue sample. A tissue sample which does not express the product is categorized as having metastatic potential.  
       [0014] Another embodiment of the invention provides a method of predicting the propensity for metastatic spread of a breast tumor preferentially to bone or lung. An expression product of a gene which comprises a sequence selected from the group consisting of SEQ ID NO:1, 5, 11, 18, 20, 24, 30, 33, 35, 36, 38, 45, 52, 58, 65, 66, 70, 74, 76, and 80 is measured in a breast tumor sample. A breast tumor sample which expresses the product is categorized as having a propensity to metastasize to bone or lung.  
       [0015] Even another embodiment of the invention provides a method of predicting propensity for metastatic spread of a breast tumor preferentially to lung. An expression product of a gene which comprises a sequence selected from the group consisting of SEQ ID NOS:2, 4, 9, 13, 14, 19, 26, 29, 39-41, 48, 55, 57, 60, 63, 64, 72, 73, 82, and 83 is measured in a breast tumor sample. A breast tumor sample which expresses the product is characterized as having a propensity to metastasize to lung.  
       [0016] Still another embodiment of the invention provides a method of predicting propensity for metastatic spread of a colon tumor. An expression product of a gene which comprises the nucleotide sequence shown in SEQ ID NO:56 is measured in a colon tumor sample. A colon tumor sample which expresses the product is characterized as having a low propensity to metastasize.  
       [0017] Even another embodiment of the invention provides a method for determining metastasis in a tissue sample. An expression product of a gene which comprises a coding sequence selected from the group consisting of SEQ ID NOS:3, 7, 8, 10, 12, 15-17, 21, 23, 25, 28, 31, 34, 37, 42-44, 46, 47, 49, 61, 62, 67, 68, 75, 77-79, 81, 84, and 85 is measured in a tissue sample. A tissue sample which expresses the product is categorized as non-metastatic.  
       [0018] Yet another embodiment of the invention provides a method for determining metastasis in a tissue sample. An expression product of a gene which comprises a coding sequence selected from the group consisting of SEQ ID NOS:3, 7, 8, 10, 12, 15-17, 21, 23, 25, 28, 31, 34, 37, 42-44, 46, 47, 49, 61, 62, 67, 68, 75, 77-79, 81, 84, and 85 is measured in a tissue sample. A tissue sample which does not express the product is categorized as metastatic.  
       [0019] The invention thus provides the art with a number of genes and proteins, which can be used as markers of metastasis. These are useful for more rationally prescribing the course of therapy for breast or colon cancer patients.  
       DETAILED DESCRIPTION  
       [0020] It is a discovery of the present invention that a number of genes are differentially expressed between metastatic cancer cells, especially cancer cells of the breast and colon, and non-metastatic cancer cells. These genes are metastatic marker genes. This information can be utilized to make diagnostic reagents specific for the expression products of the differentially expressed genes. It can also be used in diagnostic and prognostic methods which will help clinicians in planning appropriate treatment regimes for cancers, especially of the breast or colon.  
       [0021] Some of the polynucleotides disclosed herein represent novel genes which are differentially expressed between non-metastatic cancer cells and cancer cells which have a potential to metastasize. SEQ ID NOS: 1-63 represent novel metastatic marker genes (Table 1). SEQ ID NOS:64-85 represent known genes which have been found to be differentially expressed in metastatic relative to non-metastatic cancer cells (Table 2). Some of the metastatic marker genes disclosed herein are expressed in metastatic cells relative to non-metastatic cells, particularly in breast cancer cells which metastasize to bone and lung (SEQ ID NOS:1, 5, 11, 18, 20, 22, 24, 30, 33, 35, 36, 38, 45, 52, 58, 65, 66, 70, 74, 76, and 80). One metastatic marker gene (SEQ ID NO:56) is expressed in non-metastatic breast cancer cells and in colon cancer cells with low metastatic potential. Other metastatic marker genes are expressed in metastatic cancer cells, particularly in breast cancer cells which metastasize only to lung (SEQ ID NOS:2, 4, 9, 13, 14, 19, 26, 29, 39-41, 48, 55, 57, 60, 63, 64, 72. 73, 82, and 83). Still other metastatic marker genes (SEQ ID NOS:3, 7, 8, 10, 12, 15-17, 21, 23, 28, 31, 34, 37, 42-44, 46, 47, 49, 61, 62, 67, 68, 75, 77-79, 81, 84, and 85) are expressed in cancer cells which do not typically metastasize, particularly in breast cancer cells. Identification of these relationships and markers permits the formulation of reagents and methods as further described below. Other metastatic marker genes, such as those which comprise a nucleotide sequence shown in SEQ ID NOS:6, 27, 32, and 54, can be used to identify cancerous tissue, particularly breast cancer tissue.  
       [0022] Sequences of metastatic marker genes are disclosed in SEQ ID NOS. 1-85. Metastatic marker proteins can be made on expression of the disclosed polynucleotide molecules. Amino acid sequences encoded by novel polynucleotides of the invention can be predicted by running a translation program for each of three reading frames for a disclosed sequence and its complement. Complete polynucleotide sequences can be obtained by chromosome walking, screening of libraries for overlapping clones, 5′ RACE, or other techniques well known in the art.  
       [0023] Reference to metastatic marker nucleotide or amino acid sequences includes variants which have similar expression patterns in metastatic relative to non-metastatic cells, as described below. Metastatic marker polypeptide can differ in length from full-length metastatic marker proteins and contain at least 6, 8, 10, 12, 15, 18, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 140, 160, 180, or 200 or more contiguous amino acids of a metastatic marker protein.  
       [0024] Variants of marker proteins and polypeptides can also occur. Metastatic marker protein or polypeptide variants can be naturally or non-naturally occurring. Naturally occurring metastatic marker protein or polypeptide variants are found in humans or other species and comprise amino acid sequences which are substantially identical to the proteins encoded by genes corresponding to the nucleotide sequences shown in SEQ ID NOS: 1-85 or their complements. Non-naturally occurring metastatic marker protein or polypeptide variants which retain substantially the same differential expression patterns in metastatic relative to non-metastatic cancer cells as naturally occurring metastatic marker protein or polypeptide variants are also included here. Preferably, naturally or non-naturally occurring metastatic marker protein or polypeptide variants have amino acid sequences which are at least 85%, 90%. or 95% identical to amino acid sequences encoded by the nucleotide sequences shown in SEQ ID NOS:1-85. More preferably, the molecules are at least 98% or 99% identical. Percent sequence identity between a wild-type protein or polypeptide and a variant is determined by aligning the wild-type protein or polypeptide with the variant to obtain the greatest number of amino acid matches, as is known in the art, counting the number of amino acid matches between the wild-type and the variant, and dividing the total number of matches by the total number of amino acid residues of the wild-type sequence.  
       [0025] Preferably, amino acid changes in metastatic marker protein or polypeptide variants are conservative amino acid changes. i.e., substitutions of similarly charged or uncharged amino acids A conservative amino acid change involves substitution of one of a family of amino acids which are related in their side chains. Naturally occurring amino acids are generally divided into four families: acidic (aspartate, glutamate). basic (Iysine, argintine, histamine), non-polar (adenine, v7alkine. leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), and uncharged polar (glycine, asparagine, glutamine, cystine, serine, threonine, tyrosine) amino acids. Phenylalanine, tryptophan, and tyrosine are sometimes classified jointly as aromatic amino acids.  
       [0026] It is reasonable to expect that an isolated replacement of a leucine with an isoleucine or valine, an aspartate with a glutamate, a threonine with a serine, or a similar replacement of an amino acid Keith a structurally related amino acid will not have a major effect on the biological properties of the resulting metastatic marker protein or polypeptide variant. Properties and functions of metastatic marker protein or polypeptide variants are of the same type as a metastatic marker protein or polypeptide comprising amino acid sequences encoded by the nucleotide sequences shown in SEQ ID NOS:.1-85, although the properties and functions of variants can differ in degree. Whether an amino acid change results in a metastatic marker protein or polypeptide variant with the appropriate differential expression pattern can readily be determined. For example, nucleotide probes can be selected from the marker gene sequences disclosed herein and used to detect marker gene mRNA in Northern blots or in tissue sections, as is known in the art. Alternatively, antibodies which specifically bind to protein products of metastatic marker genes can be used to detect expression of metastatic marker proteins.  
       [0027] Metastatic marker variants include glycosylated forms aggregative conjugates with other molecules, and covalent conjugates with unrelated chemical moieties. Metastatic marker variants also include allergic variants, species variants, and muteins. Truncations or deletions of regions which do not affect the differential expression of metastatic marker genes are also metastatic marker variants. Covalent variants can be prepared by linking functionalities to groups which are found in the amino acid chain or at the N- or C-terminal residue, as is known in the art.  
       [0028] Full-length metastatic marker proteins can be extracted, using standard biochemical methods, from metastatic marker protein-producing human cells, such as metastatic breast or colon cancer cells. An isolated and purified metastatic marker protein or polypeptide is separated from other compounds which normally associate with a metastatic marker protein or polypeptide in a cell, such as certain proteins, carbohydrates, lipids, or subcellular organelles. A preparation of isolated and purified metastatic marker proteins or polypeptides is at least 80% pure; preferably, the preparations are 90%, 95%, or 99% pure.  
       [0029] Metastatic marker proteins and polypeptides can also be produced by recombinant DNA methods or by synthetic chemical methods. For production of recombinant metastatic marker proteins or polypeptides, coding sequences selected from the nucleotide sequences shown in SEQ ID NOS:1-85. or variants of those sequences which encode metastatic marker proteins, can be expressed in known procaryotic or eukaryotic expression systems (see below). Bacterial, yeast, insect, or mammalian expression systems can be used, as is known in the art.  
       [0030] Alternatively, synthetic chemical methods, such as solid phase peptide synthesis, can be used to synthesize a metastatic marker protein or polypeptide. General means for the production of peptides, analogs or derivatives are outlined in CHEMISTRY AND BIOCHEMISTRY OF AMINO ACIDS. PEPTIDES, AND PROTEINS—A SURVEY OF RECENT DEVELOPMENTS, Weinstein. B. ed., Mylarcell Dekker, Inc., publ., New York (1983). Moreover, substitution of D-amino acids for the normal L-stereoisomer can be carried out to increase the half-life of the molecule. Metastatic marker variants can be similarly produced.  
       [0031] Non-naturally occurring fusion proteins comprising at least 6, 8, 10, 12, 15, 18, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 140, 160, 180, or 200 or more contiguous metastatic marker amino acids can also he constructed. Human metastatic marker fusion proteins are useful for generating antibodies against metastatic marker amino acid sequences and for use in various assay systems. For example, metastatic marker fusion proteins can be used to identify proteins which interact with metastatic marker proteins and influence their functions Physical methods, such as protein affinity chromatography, or library-based assays for protein-protein interactions, such as the yeast two-hybrid or phage display systems, can also be used for this purpose. Such methods are well known in the art and can also be used as drug screens.  
       [0032] A metastatic marker fusion protein comprises two protein segments fused together by means of a peptide bond. The first protein segment comprises at least 6, 8, 10, 12, 15, 18, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 120, 140, 160, 180, or 200 or more contiguous amino acids of a metastatic marker protein. The amino acids can be selected from the amino acid sequences encoded by the nucleotide sequences shown in SEQ ID NOS: 1-85 or from variants of those sequences, such as those described above. The first protein segment can also comprise a full-length metastatic marker protein.  
       [0033] The second protein segment can be a full-length protein or a protein fragment or polypeptide. The fusion protein can be labeled with a detectable marker, as is known in the art, such as a radioactive, fluorescent, chemiluminescent, or biotinylated marker. The second protein segment can be an enzyme which will generate a detectable product, such as p-galactosidase. The first protein segment can be N-terminal or C-terminal, as is convenient.  
       [0034] Techniques for making fusion proteins, either recombinantly or by covalently linking two protein segments, are also well known. Recombinant DNA methods can be used to prepare metastatic marker fusion proteins, for example, by making a DNA construct which comprises coding sequences selected from SEQ ID NOS:1-85 in proper reading frame with nucleotides encoding the second protein segment and expressing the DNA construct in a host cell, as described below.  
       [0035] Isolated and purified metastatic marker proteins, polypeptides, variants, or fusion proteins can be used as immunogens, to obtain preparations of antibodies which specifically bind to a metastatic marker protein. The antibodies can be used, inter alia, to detect wild-tape metastatic marker proteins in human tissue and fractions thereof. The antibodies can also be used to detect the presence of mutations in metastatic marker genes which result in under- or over-expression of a metastatic marker protein or in expression of a metastatic marker protein with altered size or electrophoretic mobility.  
       [0036] Preparations of polyclonal or monoclonal antibodies can be made using standard methods. Single-chain antibodies can also be prepared. Single-chain antibodies which specifically bind to metastatic marker proteins, polypeptides, variants, or fusion proteins can be isolated, for example, from single-chain immunoglobulin display libraries, as is known in the art. The library is “panned” against metastatic marker protein amino acid sequences, and a number of single chain antibodies which bind with high-affinity to different epitomes of metastatic marker proteins can be isolated. Hayashi et al., 1995,  Gene  160:129-30. Single-chain antibodies can also be constructed using a DNA amplification method, such as the polymerase chain reaction (PCR), using hybridoma cDNA as a template. Thirion et al., 1996,  Eur. J. Cancer Prev.  5:507-11.  
       [0037] Single-chain antibodies can be mono- or bispecific, and can be bivalent or tetravalent. Construction of tetravalent, bispecific single-chain antibodies is taught in Coloma and Morrison, 1997,  Nat. Biotechnol.  15:159-63. Construction of bivalent, bispecific single-chain antibodies is taught in Mallender and Voss, 1994,  J Biol. Chem.  269:199-206.  
       [0038] A nucleotide sequence encoding the single-chain antibody can be constructed using manual or automated nucleotide synthesis, cloned into DNA expression constructs using standard recombinant DNA methods, and introduced into cells which express the coding sequence, as described below. Alternatively, single-chain antibodies can be produced directly using, for example, filamentous phage technology. Verhaar et al., 1995,  Int. J. Cancer  61:497-501; Nicholls et all. 1993.  J Immunol. Meth  165:81-91.  
       [0039] Metastatic marker-specific antibodies specifically bind to epitomes present in a full-length metastatic marker protein having an amino acid sequence encoded by a nucleotide sequence shown in SEQ ID NOS: 1-85, to metastatic marker polypeptides, or to metastatic marker variants, either alone or as part of a fusion protein. Preferably, metastatic marker epitopes are not present in other human proteins. Typically, at least 6, 8, 10, or 12 contiguous amino acids are required to form an epitope. However, epitopes which involve non-contiguous amino acids may require more, e.g. at least 15, 25, or 50 amino acids.  
       [0040] Antibodies which specifically bind to metastatic marker proteins, polypeptides, fusion proteins, or variants provide a detection signal at least 5-, 10-, or 20-fold higher than a detection signal provided with other proteins when used in Western blots or other immunochemical assays. Preferably, antibodies which specifically bind to metastatic marker epitopes do not detect other proteins in immunochemical assays and can immunoprecipitate a metastatic marker protein, polypeptide, fusion protein, or variant from solution.  
       [0041] Antibodies can be purified by methods well known in the art. Preferably, the antibodies are affinity purified, by passing the antibodies over a column to which a metastatic marker protein, polypeptide, variant, or fusion protein is bound. The bound antibodies can then be eluted from the column, for example, using a buffer with a high salt concentration.  
       [0042] Subgenomic polynucleotides contain less than a whole chromosome. Preferable, the polynucleotides are intron-free. In a preferred embodiment, the polynucleotide molecules comprise a contiguous sequence of 10, 11, 12, 15, 20, 25, 30, 32, 35, 40, 45, 50, 60, 70, 74, 80, 90, 100, 125, 150, 154, 175, 182, 200, 243, or 268 nucleotides selected from SEQ ID NOS:1-85 or the complements thereof. The complement of a nucleotide sequence shown in SEQ ID NOS:1-85 is a continuous nucleotide sequence which forms Watson-Crick base pairs with a contiguous nucleotide sequence shown in SEQ ID NOS:1-85 The complement of a nucleotide sequence shown in SEQ ID NIOS:1-85 (the antisense strand) is also a subgenomic polynucleotide, and can be used provide marker protein antisense oligonucleotides. Double-stranded polynucleotides which comprise one of the nucleotide sequences shown in SEQ ID NOS:1-85 are also subgenomic polynucleotides. Metastatic marker protein subgenomic polynucleotides also include polynucleotides which encode metastatic marker protein-specific single-chain antibodies and ribozymes, or fusion proteins comprising metastatic marker protein amino acid sequences.  
       [0043] Degenerate nucleotide sequences encoding amino acid sequences of metastatic marker protein and or variants, as well as homologous nucleotide sequences which are at least 85%, 90%, 95%, 98%, or 99% identical to the nucleotide sequences shown in SEQ ID NOS:1-85, are also metastatic marker subgenomic polynucleotides. Typically, homologous metastatic marker subgenomic polynucleotide sequences can be confirmed by hybridization under stringent conditions, as is known in the art. Percent sequence identity between wild-type and homologous variant sequences is determined by aligning the wild-type polynucleotide with the variant to obtain the greatest number of nucleotide matches, as is known in the art, counting the number of nucleotide matches between the wild-tape and the variant, and dividing the total number of matches by the total number of nucleotides of the wild-type sequence. A preferred algorithm for calculating percent identity is the Smith-Waterman homology search algorithm as implemented in MPSRCH program (Oxford Molecular) using an affine gap search with the following search parameters: gap open penalty of 10, and gap extension penalty of 1.  
       [0044] Metastatic marker subgenomic polynucleotides can be isolated and purified free from other nucleotide sequences using standard nucleic acid purification techniques For example, restriction enzymes and probes can be used to isolate polynucleotide fragments which comprise nucleotide sequences encoding a metastatic marker protein. Isolated and purified subgenomic polynucleotides are in preparations which are free or at least 90% free of other molecules.  
       [0045] Complementary DNA molecules which encode metastatic marker proteins can be made using reverse transcriptase, with metastatic marker mRNA as a template. The polymerase chain reaction (PCR) or other amplification techniques can be used to obtain metastatic marker subgenomic polynucleotides, using either human genomic DNA or cDNA as a template, as is known in the art. Alternatively, synthetic chemistry techniques can be used to synthesize metastatic marker subgenomic polynucleotides which comprise coding sequences to regions of metastatic marker proteins, single-chain antibodies, or ribozymes, or which comprise antisense oligonucleotides. The degeneracy of the genetic code allows alternate nucleotide sequences to be synthesized which will encode a metastatic marker protein comprising amino acid sequences encoded by the nucleotide sequences shown in SEQ ID NOS:1-85.  
       [0046] Purified and isolated metastatic marker subgenomic polynucleotides can be used as primers to obtain additional copies of the polynucleotides or as probes for identifying wild-type and mutant metastatic marker protein coding sequences. Metastatic marker subgenomic polynucleotides can be used to express metastatic marker mRNA, protein, polypeptides, or fission proteins and to generate metastatic marker antisense oligonucleotides and ribozymes.  
       [0047] A metastatic marker subgenomic polynucleotide comprising metastatic marker protein coding sequences can be used in an expression construct Preferably, the metastatic marker subgenomic polynucleotide is inserted into an expression plasmid (for example, the Ecdyson system, pIND. In Vitro Gene). Metastatic marker subgenomic polynucleotides can be propagated in vectors and cell lines using techniques well known in the art. Metastatic marker subgenomic polynucleotides can be on linear or circular molecules. They can be on autonomously replicating molecules or on molecules without replication sequences. They can be regulated by their own or by other regulatory sequences, as are known in the art.  
       [0048] A host cell comprising a metastatic marker expression construct can then be used to express all or a portion of a metastatic marker protein. Host cells comprising metastatic marker expression constructs can be procaryotic or eukarvotic. A variety of host cells are available for use in bacterial, yeast, insect, and human expression systems and can be used to express or to propagate metastatic marker expression constructs (see below). Expression constructs can be introduced into host cells using any technique known in the art. These techniques include transferer-polycation-mediated DNA transfer, transfection with naked or encapsulated nucleic acids, liposome-mediated cellular fusion, intracellular transportation of DNA-coated latex beads, protoplast fusion, viral infection, electroporation, and calcium phosphate-mediated transfection.  
       [0049] A metastatic marker expression construct comprises a promoter which is functional in a chosen host cell The skilled artisan can readily select an appropriate promoter from the large number of cell type-specific promoters known and used in the art. The expression construct can also contain a transcription terminator which is functional in the host cell. The expression construct comprises a polynucleotide segment which encodes all or a portion of the metastatic marker protein, variant, fusion protein, antibody, or ribozyme. The polynucleotide segment is located downstream from the promoter. Transcription of the polynucleotide segment initiates at the promoter. The expression construct can be linear or circular and can contain sequences, if desired, for autonomous replication.  
       [0050] Bacterial systems for expressing metastatic marker expression constructs include those described in Chang et al.,  Nature  (1978) 275:615, Goeddel et al.,  Nature  (1979) 281:544. Goeddel et al.,  Nucleic Acids Res.  (1980) 8:4057, EP 36,776, U.S. Pat. No. 4,551,433, deBoer et al.,  Proc. Nat&#39;l Acad Sci.  USA (1983) 80:21-25, and Siebenlist et al.  Cell  (1980) 20:269.  
       [0051] Expression systems in yeast include those described in Hinnen et al.,  Proc. Nat&#39;l Acad. Sci.  USA (1978) 7:1929; Ito et al,  J. Bacteriol.  (1983) 153:163; Kurtz et al.  Mol. Cell Biol  (1986) 6:142; Kunze et al.,  J. Basic Microbiol  (1985) 25:141; Gleeson et al.,  J Gen. Microbiol.  (1986) 132:3459, Roggenkamp et al.,  Mol. Gen. Genet.  (1986) 202:302) Das et al.  J. Bacteriol.  (1984) 158:1165; De Louvencourt et al.,  J Bacteriol.  (1983) 154:737. Van den Berg et al,  Bio/Technology  (1990) 8:135; Kunze et al.,  J. Basic Microbiol  (1985) 25:141: Creg et al.,  Mol Cell Biol.  (1985) 5:3376. U.S. Pat. Nos. 4,837,148, 4,929,555; Beach and Nurse,  Nature  (1981) 300:706; Davidow et al.  Curr Genet  (1985) 10-380. Gaillardin et al.  Curr. Genet.  (1985) 10:49. Ballance et al.  Biochem Biophys Res Commun.  (1983) 112:284-289; Tilburn et al.  Gene ( 1983) 26:205-221. Yelton et al.,  Proc. Nat&#39;l Acad. Sci  USA (1984) 81:1470-1474. Kelly and Hynes.  EMBO J.  (1985) 4:475479: EP 244.234, and WO 91/00357.  
       [0052] Expression of metastatic marker expression constructs in insects can be carried out as described in U.S. Pat. No. 4,745,051, Friesen et al (1986) “The Regulation of Baculovirus Gene Expression” in: THE MOLECULAR BIOLOGY OF BACULOVIRUS (W. Doerfler, ed.), EP 127.839. EP 155.476. and Vlak et al.  J Gen. Virol  (1988) 69:765-776, Miller et al.,  Ann. Rev. Microbiol.  (1988) 42:177. Carbonell et al,  Gene  (1988) 73:409, Maeda et al.,  Nature  (1985) 315:592-594, Lebacq-Verheyden et at.,  Mol. Cell. Biol  (1988) 8:3129; Smith et al.  Proc Nat&#39;; Acad Sci.  USA (1985) 82:8404. Miyajima et al.  Gene  (1987) 58:273, and Martin et at.,  DNA  (1988) 7.99. Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts are described in Luckow et al,  Bio/Technology  (1988) 6 47-55, Miller et al., in GENETIC ENGINEERING (Setlow. J. K. et al. eds.), Vol. 8 (Plenum Publishing. 1986), pp. 277-279, and Maeda et al.,  Nature  (1985) 315:592-594.  
       [0053] Mammalian expression of metastatic marker expression constructs can be achieved as described in Dijkema et al.,  EMBO J.  (1985) 4:761, Gorman et al.,  Proc. Nat&#39;l Acad. Sci.  USA (1982b) 79:6777. Boshart et al.  Cell  (1985) 41:521 and U.S. Pat. No. 4,399,216. Other features of mammalian expression of metastatic marker 5 expression constructs can be facilitated as described in Ham and Wallace,  Meth. Enz.  (1979) 58:44, Barnes and Sato,  Anal Biochem.  (1980) 102:255, U.S. Pat. Nos. 4,767,704, 4,657,866, 4,927,762, 4,560,655. WO 90/103430. WO 87/00195, and U.S. RE No. 30,985.  
       [0054] Subgenomic polynucleotides of the invention can also be used in gene delivery vehicles, for the purpose of delivering a metastatic marker mRNA or oligonucleotide (either with the sequence of native metastatic marker mRNA or its complement), full-length metastatic marker protein, metastatic marker fusion protein, metastatic marker polypeptide, or metastatic marker-specific ribozyme or single-chain antibody, into a cell preferably a eukaryotic cell. According to the present invention, a gene delivery vehicle can be, for example, naked plasmid DNA, a viral expression vector comprising a metastatic market subgenomic polynucleotide, or a metastatic marker subgenomic polynucleotide in conjunction with a liposome or a condensing agent.  
       [0055] In one embodiment of the invention, the gene delivery vehicle comprises a promoter and a metastatic marker subgenomic polynucleotide. Preferred promoters are tissue-specific promoters and promoters which are activated by cellular proliferation, such as the thiamine kinase and thymidylate syntheses promoters. Other preferred promoters include promoters which are activatable by infection with a virus, such as the α- and β-interferon promoters, and promoters which are activatable by a hormone, such as estrogen. Other promoters which can be used include the Moloney virus LTR, the CMV promoter, and the mouse albumin promoter.  
       [0056] A metastatic marker gene delivery vehicle can comprise viral sequences such as a viral origin of replication or packaging signal. These viral sequences can be selected from viruses such as astrovirus, coronavirus, orthomyxovirus, papovavirus. paramyxovirus, parvovirus, picomavirus, poxvirus, retrovirus, togavirus or adenovirus. In a preferred embodiment, the metastatic marker gene delivery vehicle is a recombinant retroviral vector. Recombinant retroviruses and various uses thereof have been described in numerous references including, for example, Mann et al.,  Cell  33:153, 1983, Cane and Mulligan.  Proc. Nat&#39;l Acad. Sci.  USA 81:6349, 1984. Miller et al,  Human Gene Therapy  1:5-14. 1990, U.S. Pat. Nos. 4,405,712. 4,861,719, and 4,980,289, and PCT Application Nos. WO 89/02,468, WO 89/05,349. and WO 90/02,806. Numerous retroviral gene delivery) vehicles can be utilized in the present invention, including for example those described in EP 0.415.731; WO 90/07936: WO 94/03622; WO 93/25698; WO 93/25234; U.S. Pat. No. 5,219,740: WO 9311230: WO 9310218; Vile and Hart,  Cancer Res  53:3860-3864. 1993; Vile and Hart,  Cancer Res  53:962-967. 1993: Ram et al.  Cancer Res.  53:83-88, 1993: Takamiya et al,  J Neurosci. Res.  33:493-503. 1992: Baba et al,  J Neurosurg  79:729-735. 1993 (U.S. Pat. No. 4,777,127, GB 2,200,651, EP 0,345,242 and WO91/02805).  
       [0057] Particularly preferred retroviruses are derived from retroviruses Aldrich include avian leukosis virus (ATCC Nos. VR-535 and VR-247) bovine leukemia virus (VR-1315), murine leukemia virus (MLV). mink-cell tocus-inducino virus (Koch et al.  J Vir  49:828. 1984: and Oliff et al.  J Vir  48.:542. 1983), murine sarcoma virus (ATCC Nos. VR-844. 45010 and 45016), reticuloendotheliosis virus (ATCC Nos VR-994, VR-770 and 45011). Rous sarcoma virus, Mason-Pfizer monkey virus, baboon endogenous virus, endogenous feline retrovinis (e.g., RD114). and mouse or rat gL30 sequences used as a retroviral vector. Particularly preferred strains of MLV from which recombinant retroviruses can be generated include 4070A and 1504A (Hartley and Rowe.  J. Vir  19:19, 1976). Abelson (ATCC No. VR-999). Friend (ATCC No. VR-245), Graffi (Ru et al.,  J Vir  62:4722. 1993; and Yantchev  Neoplasma  26:397, 1979). Gross (ATCC No. VR-590). Kirsten (Albino et al.  J Exp Med  164:1710. 1986), Harvey sarcoma virus (Manly et al.,  J Vir  62:3540. 1988; and Albino et al.,  J. Exp Med.  164:1710. 1986) and Rauscher (ATCC No. VR-998). and Moloney MLV (ATCC No. VR-190). A particularly preferred non-mouse retrovirus is Rous sarcoma virus. Preferred Rous sarcoma viruses include Bratislava (Manly et al.,  J. Vir  62:3540. 1988: and Albino et al.  J Exp Med  164:1710 1986). Bryan high titer (e.g., ATCC Nos. VR-334, VR-657, VR-726. VR-659. and VR-728), Bryan standard (ATCC No. VR-140), Carr-Zilber (Adgighitov et al.,  Neoplasma  27:159, 1980). Engelbreth-Holm (Laurent et al.,  Biochem Biophys Acta  908:241, 1987), Harris, Prague (e g. ATCC Nos. VR-772, and 45033), and Schmidt-Ruppin (e.g., ATCC Nos. VR-724, VR-725. VR-354) viruses.  
       [0058] Any of the above retroviruses can be readily utilized in order to assemble or construct retroviral metastatic marker gene delivery vehicles given the disclosure provided herein and standard recombinant techniques (e.g., Sambrook et al.,  Molecular Cloning. A Laboratory Manual,  2d ed. Cold Spring Harbor Laboratory Press, 1989. and Kunkle, PNAS 82:488, 1985) known in the art. Portions of retroviral  Metastatic marker  expression vectors can be derived from different retroviruses. For example, retrovector LTRs can be derived from a murine sarcoma virus, a tRNA binding site from a Rous sarcoma virus, a packaging signal from a murine leukemia virus, and an origin of second strand synthesis from an avian leukosis virus. These recombinant retroviral vectors can be used to generate transduction competent retroviral vector Is particles by introducing them into appropriate packaging cell lines (.see Ser. No. 07/800.921. filed Nov. 29, 1991). Recombinant retroviruses can be produced which direct the site-specific integration of the recombinant retroviral genome into specific regions of the host cell DNA. Such site-specific integration can be mediated by a chimeric integrase incorporated into the retroviral particle (see Ser. No 08/445.466 filed May 22, 1995). It is preferable that the recombinant viral gene delivery vehicle is a replication-defective recombinant virus.  
       [0059] Packaging cell lines suitable for use with the above-described retroviral gene delivery vehicles can be readily prepared (see Ser. No. 08/240,030, filed May 9, 1994; see also WO 92/05266) and used to create producer cell lines (also termed vector cell lines or “VCLs”) for production of recombinant viral particles. In particularly preferred embodiments of the present invention, packaging cell lines are made from human (e.g., HT1080 cells) or mink parent cell lines, thereby allowing production of recombinant retroviral gene delivery vehicles which are capable of surviving inactivation in human serum. The construction of recombinant retroviral gene delivery vehicles is described in detail in WO 91/02805. These recombinant retroviral gene delivery vehicles can be used to generate transduction competent retroviral particles by introducing them into appropriate packaging cell lines (see Ser. No. 07/800.921). Similarly, adenovirus gene delivery vehicles can also be readily prepared and utilized given the disclosure provided herein (see also Berkner.  Biotechniques  6:616-627, 1988. and Rosenfeld et al.,  Science  252:431-434, 1991, WO 93/07283, WO 93/06223, and WO 9,/07282).  
       [0060] A metastatic marker gene delivery vehicle can also be a recombinant adenoviral gene delivery vehicle Such vehicles can be readily prepared and utilized given the disclosure provided herein (see Berkner.  Biotechniques  6:616. 1988. and l0 Rosenfeld et al.,  Science  252:431, 1991, WO 93/07283. WO 93/06223, and WO 93/07282). Adeno-associated viral metastatic marker gene delivery vehicles can also be constructed and used to deliver metastatic marker amino acids or nucleotides. The use of adeno-associated viral gene delivery vehicles In vitro is described in Chatterjee et al.  Science  258:1485-1488 (1991) Walsh et al.  Proc Nat&#39;l Acad. Sci  89:7257-7261 is (1992). Walsh et al.  J Clin Invest  94:1440-1448 (1994). Flotte et al.  J Biol Chem  268:3781-3790 (1993). Ponnazhagan et al.  J. Exp. Med.  1. 9:733-738 (1994). Miller et al.  Proc. Nat&#39;l Acad Sci  91. 10183-10187 1994). Einerhand et al.  Gene Ther  2:336-343 (1995). Luo et al,  Exp. Hematol  23:1261-1267 (1995), and Zhou et al.,  Gene Therapy  3. 223-279 (1996). In vito use of these vehicles is described in Flotte et al.  Proc. Nat&#39;l Acad Sci  90:10613-10617 (1993), and Kaplitt et al.  Nature Genet. ( 8:148-153 (1994).  
       [0061] In another embodiment of the invention, a metastatic marker gene delivery vehicle is derived from a togavirus. Preferred togaviruses include alphaviruses, in particular those described in U.S. Ser. No. 08/405,627. filed Mar. 15. 1995, WO 95/07994. Alpha viruses, including Sindbis and ELVS viruses can be gene deliver% vehicles for metastatic marker polynucleotides. Alpha viruses are described in WO 94/21792. WO 92/10578 and WO95/07994. Several different alphavirus gene delivery vehicle systems can be constructed and used to deliver metastatic marker subgenomic polynucleotides to a cell according to the present invention. Representative examples of such systems include those described in U.S. Pat. Nos. 5,091,309 and 5,217,879. Particularly preferred alphavirus gene delivery vehicles for use in the present invention include those which are described in WO 95/07994, and U.S. Ser. No. 08/405,627.  
       [0062] Preferably, the recombinant viral vehicle is a recombinant alphavirus viral vehicle based on a Sindbis virus. Sindbis constructs, as well as numerous similar constructs, can be readily prepared essentially as described in U.S. Ser. No. 08/198,450. Sindbis viral gene delivery vehicles typically comprise a 5′ sequence capable of initiating Sindbis virus transcription, a nucleotide sequence encoding Sindbis non-structural proteins, a viral junction region inactivated so as to prevent subgenomic fragment transcription, and a Sindbis RNA polymerase recognition sequence. Optionally, the viral junction region can be modified so that subgenomic polynucleotide transcription is reduced, increased, or maintained. As will be appreciated by those in the art, corresponding regions from other alphaviruses can be used in place of those described above.  
       [0063] The viral junction region of an alphavirus-derived gene delivery vehicle can comprise a first viral junction region which has been inactivated in order to prevent transcription of the subgenomic polynucleotide and a second viral junction region which has been modified such that subgenomic polynucleotide transcription is reduced. An alphavirus-derived vehicle can also include a 5′ promoter capable of initiating synthesis of viral RNA from cDNA and a 3′ sequence which controls transcription termination.  
       [0064] Other recombinant togaviral gene delivery vehicles which can be utilized in the present invention include those derived from Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Middleberg virus (ATCC VR-370). Ross River virus (ATCC VR-373: ATCC VR-1246), Venezuelan equine encephalitis virus (ATCC VR923: ATCC VR-1250; ATCC VR-1249. ATCC VR-539. ). and those described in U.S. Pat. Nos. 5,091,309 and 5,217,879 and in WO 92/10578. The Sindbis vehicles described above, as well as numerous similar constructs, can be readily prepared essentially as described in U.S. Ser. No. 08/198,450.  
       [0065] Other viral gene delivery vehicles suitable for use in the present invention include, for example, those derived from poliovirus (Evans et al.,  Nature  339:385, 1989, and Sabin et al.  J. Biol. Standardization  1:115, 1973) (ATCC VR-58): rhinovirus (Arnold et al.,  J. Cell. Biochem.  L401, 1990) (ATCC VR-110); pox viruses. such as canary pox virus or vaccinia virus (Fisher-Hoch et al.,  PNAS  86:317, 1989; Flexner et al.,  Ann. N Y. Acad. Sci.  569:86. 1989: Flexner et al,  Vaccine  8:17.1990; U.S. Pat. Nos. 4,603,119 and 4.769,330; WO 89/01973) (ATCC VR-111; ATCC VR-2010); SV40 (Mulligan et al.,  Nature  277 108. 1979) (ATCC VR-305), (Madzak et al.,  J. Gen. Vir.  73:1533, 1992); influenza virus (Luytjes et al.  Cell  59:1107, 1989: McMicheal et al.,  The New England Journal of Medicine  309:13, 1983; and Yap et al.,  Nature  273:238, 1978) (ATCC VR-797); parvovirus such as adeno-associated virus (Samulski et al.,  J. Vir.  63:38292 1989, and Mendelson et al.  Virology  166:154, 1988) (ATCC VR-645); herpes simplex virus (Kit et al.,  Adv Exp Med. Biol  215:219. 1989) (ATCC VR-977, ATCC VR-260):  Nature  27l: 108. 1979), human immunodeficiency virus (EPO 386.882, Buchschacher et al.  J Vir  66:2731. 1992): measles virus (EPO 440.219) (ATCC VR-24) A (ATCC VR-67: ATCC VR-1247). Aura (ATCC VR368) is Bebaru virus (ATCC VR-600: ATCC VR-1240). Cabassou (ATCC VR-922). Chikungunya virus (ATCC VR-64ATCC VR-141), Fort, Llorgaan (AFCC VR-924). Getah virus (ATCC VR-369: ATCC VR-143). Kyzlagacl (ATCC VR-927). Mayaro (ATCC VR-66). Mucambo virus (ATCC VR-580: ATCC VR-1244), Ndumu (ATCC VR-371), Pixuna virus (ATCC VR-372: ATCC VR-1245), Tonate (ATCC VR-925). Triniti (ATCC VR-469), Una (ATCC VR-374), Whataroa (ATCC VR-926). Y-62-33 (ATCC VR-375). O&#39;Nyong virus, Eastern encephalitis virus (ATCC VR-65; ATCC VR-1242), Western encephalitis virus (ATCC VR-70: ATCC VR-1251: ATCC VR-622: ATCC VR-1252), and coronavirus (Hamre et al.  Proc Soc Exp Biol Med  121:190. 1966) (ATCC VR-740).  
       [0066] A subgenomic metastatic marker polynucleotide of the invention can also be combined with a condensing agent to form a gene delivery vehicle. In a preferred embodiment, the condensing agent is a polycation, such as polylysine, polyarginine, polyomithine, protamine, spermine, spermidine, and putrescine. Many suitable methods for making such linkages are known in the art (see, for example, Ser. No. 08/366,787. filed Dec. 30. 1994).  
       [0067] In an alternative embodiment, a metastatic marker subgenomic olynucleotide is associated with a liposome to form a gene delivery vehicle. Liposomes are small, lipid vesicles comprised of an aqueous compartment enclosed by a lipid bilayer, typically spherical or slightly elongated structures several hundred Angstroms in diameter. Under appropriate conditions, a liposome can fuse with the plasma membrane of a cell or with the membrane of an endocytic vesicle within a cell which has internalized the liposome, thereby releasing its contents into the cytoplasm. Prior to interaction with the surface of a cell, however, the liposome membrane acts as a relatively impermeable barrier which sequesters and protects its contents, for example, from degradative enzymes. Additionally, because a liposome is a synthetic structure, specially designed liposomes can be produced which incorporate desirable features. See Stryer.  Biochemistry. pp.  236-240, 1975 (W. H. Freeman. San Francisco. Calif.): Szoka et al.,  Biochim. Biophys. Acta  600:1. 1980; Bayer et al,  Biochim Biophys. Acta  550:464. 1979: Rivnay et al.  Meth. Enzymol  149:119, 1987; Wang et. al.  PNAS  84:7851, 1987. Plant et al.  Anal. Biochem  176:420. 1989. and U.S. Pat. No. 4,762,915 Liposomes can encapsulate a variety of nucleic acid molecules including DNA. RNA, plasmids and expression constructs comprising metastatic marker subgenomic polynucleotides such those disclosed in the present invention.  
       [0068] Liposomal preparations for use in the present invention include cationic (positively charged), anionic (negatively charged) and neutral preparations. Cationic liposomes have been shown to mediate intracellular delivery of plasmid DNA (Felgner et al.,  Proc Nat&#39;l Acad. Sci.  USA 84:7413-7416, 1987), mRNA (Malone et al.  Proc Nat&#39;l Acad. Sci.  USA 86:6077-6081. 1989), and purified transcription factors (Debs et al.,  J. Biol. Chen.  265:10189-10192. 1990), in functional form. Cationic liposomes are readily available. For example, N[-1-2,3-dioleyloxy)propyl]-N,N.N-triethylammonium (DOTMA) liposomes are available under the trademark Lipofectin, from GIBCO BRL, Grand Island. N.Y. See also Fetgner et al.  Proc Nat&#39;l Acad. Sci.  USA 91 5148-5152.87. 1994. Other commercially available liposomes include Transfectace (DDABIDOPE) and DOTAP,IDOPE (Boerhinger). Other cationic liposomes can be prepared from readily available materials using techniques well known in the art. See, e.g, Szoka et al,.  Proc. Nat&#39;l Acad. Sci  USA 75:4194-4198. 1978; and WO 90/11092 for descriptions of the synthesis of DOTAP (1,2-bis(oleoyloxy)-3-(trimethylammonio)propane) liposomes.  
       [0069] Similarly, anionic and neutral liposomes are readily available, such as from Avanti Polar Lipids (Birmingham, Ala.). or can be easily prepared using readily available materials. Such materials include phosphatidyl choline, cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline (DOPC). dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidyl ethanolamine (DOPE), among others. These materials can also be mixed with the DOTMA and DOTAP starting materials in appropriate ratios. Methods for making liposomes using these materials are well known in the art.  
       [0070] The liposomes can comprise multilammelar vesicles (MLVs), small unilamellar vesicles (SUVs), or large unilamellar vesicles (LUVs). The various liposome-nucleic acid complexes are prepared using methods known in the art. See, e.g. Straubinger et al. METHODS OF IMMUNOLOGY (1983). Vol. 101, pp. 512-527, Szoka et al.  Proc. Nat&#39;l Acad Sci  USA 3410-3414. 1090: Papahadjopoulos et al.  Biochim. Biophys Acta  394:483. 1975: Wilson et al.  Cell  17:77, 1979; Deamer and Bangham.  Biochim Biophys Acta  443:629. 1976;Ostro et al.  Biochem Biophys. Res Commun.  76:836. 1977: Fraley et al.  Proc. Nat&#39;l Acad Sci.  USA 76:3348, 1979; Enoch and Strittmatter.  Proc. Nat&#39;l Acad. Sci  USA 76:145, 1979: Fraley et al.  J Biol Chem.  255:10431. 1980; Szoka and Papahadjopoulos.  Proc Nat&#39;l Acad. Sci.  USA 75:145. 1979: and Schaefer-Ridder et al,  Science  215:166, 1982.  
       [0071] In addition, lipoproteins can be included with a metastatic marker subgenomic polynucleotide for deliver, to a cell. Examples of such lipoproteins include chylomicrons, HDL, IDL, LDL, and VLDL. Mutants, fragments, or fusions of these proteins can also be used. Modifications of naturally occurring lipoproteins can also be used, such as activated LDL. These lipoproteins can target the delivery of polynucleotides to cells expressing lipoprotein receptors. Preferably, if lipoproteins are included with a polynucleotide, no other targeting ligand is included in the composition.  
       [0072] In another embodiment, naked metastatic marker subgenomic polynucleotide molecules are used as gene delivery vehicles, as described in WO 90/11092 and U.S. Pat. 5,580,859. Such gene delivery vehicles can be either metastatic marker DNA or RNA and, in certain embodiments, are linked to killed adenovirus. Curiel et al.,  Hum. Gene. Ther.  3:147-154. 1992. Other suitable vehicles include DNA-ligand (Wu et al.,  J. Biol. Chem.  264:16985-16987, 1989), lipid-DNA combinations (Feigner et al.,  Proc. Nat&#39;l Acad. Sci.  USA 84:7413 7417, 1989), liposomes (Wang et al.,  Proc. Nat&#39;l Acad. Sci.  84:7851-7855. 1987) and microprojectiles (Williams et al.,  Proc. Nat&#39;l Acad. Sci.  88:2726-2730, 1991).  
       [0073] One can increase the efficiency of naked metastatic marker subgenomic polynucleotide uptake into cells by coating the polynucleotides onto biodegradable latex beads. This approach takes advantage of the observation that latex beads, when incubated with cells in culture, are efficiently transported and concentrated in the perinuclear region of the cells. The beads will then be transported into cells when injected into muscle. Metastatic marker subgenomic polynucleotide-coated latex beads will be efficiently, transported into cells after endocytosis is initiated by the latex beads and thus increase gene transfer and expression efficiency. This method can be improved further by treating the beads to increase their hydrophobicity, thereby faicilitating the disruption of the endosome and release of metastatic marker subgenomic polynucleotides into the cytoplasm.  
       [0074] The invention provides a method of detecting metastatic marker gene expression in a biological sample. Detection of metastatic marker gene expression is useful, for example, for identifying metastases or for determining metastatic potential in a tissue sample, preferably a tumor. Appropriate treatment regimens can then be designed for patients who are at risk for developing metastatic cancers in other organs of the body.  
       [0075] The body sample can be, for example, a solid tissue or a fluid sample. Protein or nucleic acid expression products can be detected in the body sample. In one embodiment, the body sample is assayed for the presence of a metastatic marker protein. A metastatic marker protein comprises a sequence encoded by a nucleotide sequence shown in SEQ ID NOS:1-85 or its complement and can be detected using the marker protein-specific antibodies of the present invention. The antibodies can be labeled, for example, with a radioactive, fluorescent, biotinylated, or enzymatic tag and detected directly, or can be detected using indirect immunochemical methods, using a labeled secondary antibody. The presence of the metastatic marker proteins can be assayed, for example, in tissue sections by immunocytochemistry, or in lysates, using Western blotting, as is known in the art.  
       [0076] In another embodiment, the body sample is assayed for the presence of marker protein mRNA. A sample can be contacted with a nucleic acid hybridization probe capable of hybridizing with the mRNA corresponding the selected polypeptide. Still further, the sample can be subjected to a Northern blotting technique to detect mRNA, indicating expression of the polypeptide. For those techniques in which mRNA is detected, the sample can be subjected to a nucleic acid amplification process whereby the mRNA molecule or a selected part thereof is amplified using appropriate nucleotide primers. Other RNA detection techniques can also be used, including, but not limited to, in sito hybridization.  
       [0077] Marker protein-specific probes can be generated using the cDNA sequences disclosed in SEQ ID NOSS: 1-85. The probes are preferably at least 15 to 50 nucleotides in length, although they can be at least 8, 10, 11, 12, 20, 25, 30, 35, 40, 45, 60, 75, or 100 or more nucleotides in length. The probes can be synthesized chemically or can be generated from longer polynucleotides using restriction enzymes. The probes can be labeled, for example, with a radioactive, biotinylated, or fluorescent tag.  
       [0078] Optionally, the level of a particular metastatic marker expression product in a body sample can be quantitated. Quantitation can be accomplished, for example, by comparing the level of expression product detected in the body sample with the amounts of product present in a standard curve. A comparison can be made visually or using a technique such as densitometrn, with or without computerized assistance. For use as controls, body samples can be isolated from other humans, other non-cancerous organs of the patient being tested, or non-metastatic breast or colon cancer from the patient being tested.  
       [0079] Polynucleotides encoding metastatic marker-specific reagents of the invention, such as antibodies and nucleotide probes, can be supplied in a kit for detecting marker gene expression products in a biological sample. The kit can also contain buffers or labeling components, as well as instructions for using the reagents to detect the marker expression products in the biological sample.  
       [0080] If expression of a metastatic marker gene having a nucleotide sequence shown in SEQ ID NOS:2, 4, 9, 13, 14, 19, 26, 29, 39-41, 48, 55, 57, 60, 63, 64, 72, 73, 82, or 83 is detected, the biological sample contains cancer cells which will likely metastasize to the lung. If expression of a metastatic marker gene having a nucleotide sequence shown in SEQ ID NOS:1, 5, 11, 18, 20, 22, 24, 30, 33, 35, 36, 38, 45, 52, 58, 65, 66, 70, 74, 76, or 80 is detected, the biological sample contains cancer cells which will likely metastasize to the bone and/or lung. On the other hand, if expression of a metastatic marker gene having a nucleotide sequence shown in SEQ ID NOS:3, 7, 8, 10, 19, 15-17, 21, 23, 25, 28, 31, 34. 37, 42-44, 46, 47, 49-51, 53, 59, 61, 62, 67, 68, 75, 77-79, 81, 84, or 85 is detected, the biological sample contains cancer cells which will likely not metastasize. Detection of expression of a tastatic maiker gene comprising the nucleotide sequence shown in SEQ ID NO:56 also indicates that the biological sample contains cancer cells which will likely metastasize. This information can be used, for example, to design treatment regimens. Treatment regiments can include altering expression of one or more metastatic marker genes, as desired. Metastatic marker gene expression can be altered for therapeutic purposes, as described below, or can be used to identify therapeutic agents.  
       [0081] In one embodiment of the invention, expression of a metastatic marker gene whose expression is up-regulated in metastatic cancer is decreased using a ribozyme, an RNA molecule with catalytic activity. See. e.g. Cech. 1987,  Science  236:1532-1539; Cech, 1990,  Ann Rev. Biochem.  59:543-568; Cech, 1992,  Curr Opin Struct Biol.  2:605-609: Couture and Stinchcomb, 1996.  Trends Genet.  12:510-515. Ribozymes can be used to inhibit gene function by cleaving an RNA sequence, as is known in the art (e g. Haseloff et al., U.S. Pat. No. 5,641,673).  
       [0082] Coding sequences of metastatic marker genes can be used to generate ribozymes which will specifically bind to mRNA transcribed from a metastatic marker gene. Methods of designing and constructing ribozymes which can cleave other RNA molecules in trans in a highly sequence specific manner have been developed and described in the art (see Haseloff. J. et al. (1988),  Nature  334:585-591). For example, the cleavage activity of ribozymes can be targeted to specific RNAs by engineering a discrete “hybridization” region into the ribozyme. The hybridization region contains a sequence complementary to the taroet RNA and thus specifically hybridizes with the target (see, for example, Gerlach. W. L. et al., EP 321.201). Longer complementary sequences can be used to increase the affinity of the hybridization sequence for the target. The hybridizing and cleavage regions of the ribozyme can be integrally related; thus, upon hybridizing to the target RNA through the complementary regions, the catalytic region of the ribozyme can cleave the target.  
       [0083] Ribozymes can be introduced into cells as part of a DNA construct, as is known in the art. The DNA construct can also include transcriptional regulatory elements, such as a promoter element, an enhancer or IJAS element, and a transcriptional terminator signal, for controlling the transcription of the ribozyme in the cells.  
       [0084] Mechanical methods, such as microinjection, liposome-mediated transfection, electroporation, or calcium phosphate precipitation, can be used to introduce a ribozyme-containing DNA construct into cells whose division it is desired to decrease, as described above. Alternatively, if it is desired that a DNA construct be stably retained by the cells, the DNA construct can be supplied on a plasmid and maintained as a separate element or integrated into the genome of the cells, as is known in the art.  
       [0085] As taught in Haseloff et al., U.S. Pat. No. 5,641,673. ribozymes can be engineered so that their expression sill occur in response to factors which induce expression of metastatic marker genes. Ribozymes can also be engineered to provide an additional level of regulation, so that destruction of mRNA occurs only when both a ribozyme and a metastatic marker gene are expressed in the cells.  
       [0086] Expression of a metastatic marker gene can also be altered using an antisense oligonucleotide sequence The antisense sequence is complementary to at least a portion of the coding sequence of a metastatic marker gene having a nucleotide sequence shown in SEQ ID NOS: 1-85. The complement of a nucleotide sequence shown in SEQ ID NOS: 1-85 is a contiguous sequence of nucleotides which form Watson-Crick basepairs with a contiguous nucleotide sequence shown in SEQ ID NOS: 1-85.  
       [0087] Preferably, the antisense oligonucleotide sequence is at least six nucleotides in length, but can be at least about 8, 12, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides long. Longer sequences can also be used. Antisense oligonucleotide molecules can be provided in a DNA construct and introduced into cells whose division is to be decreased, as described above.  
       [0088] Antisense oligonucleotides can comprise deoxyriboiiucleotides, ribonucleotides, or a combination of both oligonucleotides can be synthesized manually or by an automated synthesizer, covalently linking the 5′ end of one nucleotide with the 3′ end of another nucleotide with non-phosphodiester internucleotide linkages such alkylphosphonates, phosphorothioates, phosphorodithioates, alkylphosphonothioates, alkylphosphonates, phosphoramidates. phosphate esters, carbamates, acetamidate, carboxymethvl esters, carbonates, and phosphate triesters. See Brown, 1994.  Meth Mo. Biol.  20:1-8; Sonveaux, 1994,  Meth. Mol. Biol.  26:1-72; Uhlmann et al., 1990.  Chem. Rev.  90:543-583.  
       [0089] Although precise complementary is not required for successful duplex formation between an antisense molecule and the complementary coding sequence of a metastatic marker gene, antisense molecules with no more than one mismatch are preferred. One skilled in the art can easily use the calculated melting point of a metastatic marker gene antisense-sense pair to determine the degree of mismatching which will be tolerated between a particular antisense oligonucleotide and a particular coding sequence of the selected gene.  
       [0090] Antisense oligonucleotides can be modified without affecting their ability to hybridize to a metastatic marker protein coding sequence. These modifications can be internal or at one or both ends of the antisense molecule. For example, intemucleoside phosphate linkages can be modified by adding cholesteryl or diamine moieties with varying numbers of carbon residues between the amino groups and terminal ribose. Modified bases and/or sugars, such as arabinose instead of ribose, or a 3′,5′-substituted oligonucleotide in which the 3′ hydroxyl group or the 5′ phosphate group are substituted, can also be employed in a modified antisense oligonucleotide. These modified oligonucleotides can be prepared by methods well known in the art. Agrawal et al., 1992. Trends Biotechnol. 10:152-158; Uhlmann et al., 1990,  Chem. Rev.  90:543-584; Uhlmann et al., 1987.  Tetrahedron. Lett.  215:3539-3542.  
       [0091] Antibodies of the invention which specifically bind to a metastatic marker protein can also be used to alter metastatic marker gene expression. By antibodies is meant antibodies and parts or derivatives thereof, such as single chain antibodies, that retain specific binding for the protein. Specific antibodies bind to metastatic marker proteins and prevent the proteins from functioning in the cell. Polynucleotides encoding specific antibodies of the invention can be introduced into cells, as described above.  
       [0092] Marker proteins of the present invention can be used to screen for drugs which have a therapeutic anti-metastatic effect. The effect of a test compound on metastatic marker protein synthesis can also be used to identify test compounds which modulate metastasis. Test compounds which can be screened include any substances, whether natural products or synthetic, which can be administered to the subject. Libraries or mixtures of compounds can be tested. The compounds or substances can be those for which a pharmaceutical effect is previously known or unknown.  
       [0093] A cell is contacted with a test compound. The cell can be any cell, such as a colon cancer cell, which ordinarily synthesizes the metastatic marker protein being measured. For example, Tables 1 and 2 provide appropriate cell types which can be used for screening assays.  
       [0094] Synthesis of metastatic marker proteins can be measured by any means for measuring protein synthesis known in the art, such as incorporation of labeled amino acids into proteins and detection of labeled metastatic marker proteins in a polyacrylamide gel. The amount of metastatic marker proteins can be detected, for example, using metastatic marker protein-specific antibodies of the invention in Western blots. The amount of the metastatic marker proteins synthesized in the presence or absence of a test compound can be determined by any means known in the art, such as comparison of the amount of metastatic marker protein synthesized with the amount of the metastatic marker proteins present in a standard curve.  
       [0095] The effect of a test compound on metastatic marker protein synthesis can also be measured by Northern blot analysis, by measuring the amount of metastatic marker protein mRNA expression in response to the test compound using metastatic marker protein specific nucleotide probes of the invention, as is known in the art.  
       [0096] Typically, biological sample is contacted with a range of concentrations of the test compound, such as 1.0 nM, 5.0 nM, 10 nM, 50 nM, 100 nM, 500 nM, 1 mM, 10 mM, 50 mM, and 100 mM. Preferably, the test compound increases or decreases expression of a metastatic marker protein by 60%, 75%, or 80%. More preferably, an increase or decrease of 85%, 90%, 95%, or 98% is achieved.  
       [0097] The invention provides compositions for increasing or decreasing expression of metastatic marker protein. Therapeutic compositions for increasing metastatic marker gene expression are desirable for markers which are down-regulated in metastatic cells These compositions comprise polynucleotides encoding all or at least a portion of a metastatic marker protein gene expression product. Preferably, the therapeutic composition contains an expression construct comprising a promoter and a polynucleotide segment encoding at least a portion of the metastatic marker protein which is effective to increase or decrease metastatic potential. Portions of metastatic marker genes or proteins which are effective to decrease metastatic potential of a cell can be determined, for example, by introducing various portions of metastatic marker genes or polypeptides into metastatic cell lines, such as MDA-MB-231, MDA-MB-435, Km12C, or Km12L4. and assaying the division rate of the cells or the ability of the cells to form metastases when implanted in vivo, as is known in the art. Non-metastatic cell lines, such as MCF-7, can be used to assay the ability of a portion of a metastatic marker protein to increase expression of a metastatic marker gene.  
       [0098] Within the expression construct, the polynucleotide segment is located downstream from the promoter, and transcription of the polynucleotide segment initiates at the promoter. A more complete description of gene transfer vectors, especially retroviral vectors is contained in U.S. Ser. No. 08/869,309. which is incorporated herein by reference.  
       [0099] Decreased metastatic marker gene expression is desired in conditions in which the marker gene is up-regulated in metastatic cancer. Therapeutic compositions for treating these disorders comprise a polynucleotide encoding a reagent which specifically binds to a metastatic marker protein expression product, as disclosed herein.  
       [0100] Metastatic marker therapeutic compositions of the invention can comprise a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known to those in the art. Such carriers include, but are not limited to, large, slowly metabolized macromolecules, such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactie virus particles. Pharmaceutically acceptable salts can also be used in the composition, for example, mineral salts such as hydrochlorides, hydrobromides, phosphates, or sulfates, as well as the salts of organic acids such as acetates, proprionates, malonates, or benzoates.  
       [0101] Therapeutic compositions can also contain liquids, such as water, saline, glycerol, and ethanol, as well as substances such as wetting agents, emulsifying agents, or pH buffering agents. Liposomes, such as those described in U.S. Pat. No. 5,422,120, WO 95/13796, WO 91/14445. or EP 524,968 B1, can also be used as a carrier for the therapeutic composition.  
       [0102] Typically, a therapeutic metastatic marker composition is prepared as an injectable, either as a liquid solution or suspension; however, solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection can also be prepared. A metastatic marker composition can also be formulated into an enteric coated tablet or gel capsule according to known methods in the art, such as those described in U.S. Pat. No. 4,853,230, EP 225,189, AU 9,224,296, and AU 9,230,801,  
       [0103] Administration of the metastatic marker therapeutic agents of the invention can include local or systemic administration, including injection, oral administration, particle gun, or catheterized administration, and topical administration. Various methods can be used to administer a therapeutic metastatic marker composition directly to a specific site in the body.  
       [0104] For treatment of tumors, including metastatic lesions, for example, a therapeutic metastatic marker composition can be injected several times in several different locations within the body of tumor. Alternatively, arteries which serve a tumor can be identified, and a therapeutic composition injected into such an artery, in order to deliver the composition directly into the tumor.  
       [0105] A tumor which has a necrotic center can be aspirated and the composition injected directly into the now empty center of the tumor. A therapeutic metastatic marker composition can be directly administered to the surface of a tumor, for example, by topical application of the composition, X-ray imaging can be used to assist in certain of the above delivers; methods. Combination therapeutic agents, including a metastatic marker proteins or polypeptide or a metastatic nature subgenomic polynucleotide and other therapeutic agents, can be administered simultaneously or sequentially.  
       [0106] Receptor-mediated targeted delivery can be used to deliver therapeutic compositions containing metastatic marker subgenomic polynucleotides, proteins, or reagents such as antibodies, ribozymes, or antisense oligonucleotides to specific tissues. Receptor-mediated delivery techniques are described in, for example, Findeis et al. (1993),  Trends in Biotechnol  11, 902-05; Chiou et al. (1994). GENE THERAPEUTICS METHODS AND APPLICATIONS OF DIRECT GENE TRANSFER (J. A. Wolff, ed.); Wu &amp; Wu (1988),  J. Biol. Chem.  263, 691-24; Wu et al. (1994),  J. Biol. Chem.  269, 542-46; Zenke et al. (1990).  Proc. Nat&#39;l Acad. Sci  USA. 87, 3655-59; Wu et al. (1991).  J. Biol Chem.  266, 338-42.  
       [0107] Alternatively, a metastatic marker therapeutic composition can be introduced into human cells ex vivo, and the cells then replaced into the human. Cells can be removed from a variety of locations including, for example, from a selected tumor or from an affected organ. In addition, a therapeutic composition can be inserted into non-affected, for example, dermal fibroblasts or peripheral blood leukocytes. If desired, particular fractions of cells such as a T cell subset or stem cells can also be specifically removed from the blood (see, for example, PCT WO 91/16116). The removed cells can then be contacted with a metastatic marker therapeutic composition utilizing any of the above-described techniques, followed by the return of the cells to the human, preferably to or within the vicinity of a tumor or other site to be treated. The methods described above can additionally comprise the steps of depleting fibroblasts or other non-contaminating tumor cells subsequent to removing tumor cells from a human, and/or the step of inactivating the cells, for example, by irradiation.  
       [0108] Both the dose of a metastatic marker composition and the means of administration can be determined based on the specific qualities of the therapeutic composition, the condition, age, and weight of The patient, the progression of the disease, and other relevant factors. Preferably, a therapeutic composition of the invention increases or decreases expression of the metastatic marker genes by 50%, 60%, 70%, or 80%. Most preferably, expression of the metastatic matier genes is increased or decreased by 90%, 95%, 99%, or 100%. The eftectiveness of the mechanism chosen to alter expression of the metastatic marker genes can be assessed using methods well known in the art, such as hybridization of nucleotide probes to mRNA of the metastatic marker genes, quantitative RT-PCR, or detection of an the metastatic marker proteins using specific antibodies of the invention.  
       [0109] If the composition contains the metastatic marker proteins, polypeptide. Or antibody, effective dosages of the composition are in the range of about 5 μg to about 50 μg/kg of patient body weight, about 50 μg to about 5 mg/kg, about 100 μg to about 500 μg/kg of patient body weight, and about 200 to about 250 μg/kg.  
       [0110] Therapeutic compositions containing metastatic marker subgenomic polynucleotides can be administered in a range of about 100 ng to about 200 mg of DNA for local administration. Concentration ranges of about 500 ng to about 50 mg, about 1 μg to about 2 mg, about 5 μg to about 500 μg, and about 20 μg to about 100 μg of DNA can also be used during a gene therapy protocol. Factors such as method of action and efficacy of transformation and expression are considerations that will affect the dosage required for ultimate efficacy of the metastatic marker subgenomic polynucleotides. Where greater expression is desired over a larger area of tissue, larger amounts of metastatic marker subgenomic polynucleotides or the same amounts readministered in a successive protocol of administrations, or several administrations to different adjacent or close tissue portions of, for example, a tumor site, can be required to effect a positive therapeutic outcome. In all cases, routine experimentation in clinical trials will determine specific ranges for optimal therapeutic effect.  
       [0111] Expression of an endogenous metastatic marker gene in a cell can also be altered by introducing in frame with the endogenous metastatic marker gene a DNA construct comprising a metastatic marker protein targeting sequence, a regulatory sequence, an exon, and an unpaired splice donor site by homologous recombination, such that a homologously recombinant cell comprising the DNA construct is formed The new transcription unit can be used to turn the inetastatic marker gene on off as desired. This method of affecting endogenous gene expression is taught in U.S. Pat. No. 5,641,670, which is incorporated herein by reference.  
       [0112] The targeting sequence is a segment of at least 10, 12, 15, 20, or 50 contiguous nucleotides selected from the nucleotide sequences shown in SEQ ID NOS:1-85 or the complements thereof. The transcription unit is located upstream of a coding sequence of the endogenous metastatic marker protein gene. The exogenous regulatory sequence directs transcription of the coding sequence of the metastatic marker genes.  
       [0113] A metastatic marker subgenomic polynucleotide can also be delivered to subjects for the purpose of screening test compounds for those which are useful for enhancing transfer of metastatic marker subgenomic polynucleotides to the cell or for enhancing subsequent biological effects of metastatic marker subgenomic polynucleotides within the cell. Such biological effects include hybridization to complementary metastatic marker mRNA and inhibition of its translation, expression of a metastatic marker subgenomic polynucleotide to form metastatic marker mRNA and/or metastatic marker protein, and replication and integration of a metastatic marker subgenomic polynucleotide. The subject can be a cell culture or an animal, preferably a mammal, more preferably a human.  
       [0114] Test compounds which can be screened include any substances, whether natural products or synthetic, which can be administered to the subject. Libraries or mixtures of compounds can be tested. The compounds or substances can be those for which a pharmaceutical effect is previously known or unknown. The compounds or substances can be delivered before after, or concomitantly with a metastatic marker subgenomic polynucleotide. They can be administered separately or in admixture with a metastatic marker subgenomic polynucleotide.  
       [0115] Integration of a delivered metastatic marker subgenomic polynucleotide can be monitored by any means known in the art. For example, Southern blotting of the delivered metastatic marker subgenomic polynucteotide can be performed. A change in the size of the fragments of a delivered polynucleotide indicates integration. Replication of a delivered polynucleotide can be monitored inter alia by detecting incorporation of labeled nucleotides combined with hybridization to a metastatic marker probe. Expression of metastatic marker suboenomic polynucleotide can be monitored by detecting production of metastatic marker mRNA which hybridizes to the delivered polynucleotide or by detecting metastatic marker protein. Metastatic marker protein can be detected immunologically. Thus, the delivery of metastatic marker subgenomic polynucleotides according to the present invention provides an excellent system for screening test compounds for their ability to enhance transfer of metastatic marker subgenomic polynucleotides to a cell, by enhancing delivery, integration, hybridization, expression, replication or integration in a cell in vitro or in an animal, preferably a mammal, more preferably a human.  
       [0116] The above disclosure generally describes the present invention. A more complete understanding can be obtained by reference to the following specific examples which are provided herein for purposes of illustration only, and are not intended to limit the scope of the invention.  
     
    
    
     EXAMPLE 1  
     [0117] DIFFERENTIALLY EXPRESSED GENES  
     [0118] This example demonstrates polynucleotides that are differentially expressed in human breast or colon cancer cell lines.  
     [0119] Human cell lines used to identify differentially expressed polynucleotides are the human breast cancer cell lines MCF-7 (non-metastatic), MDA-MB-23 1 (metastatic to bone and/or lung), and MDA-MB-435 (metastatic to lung) (Brinkley and Cailleau, 1980,  Cancer Res.  40:3118), and the colon cancer cell lines Km12C (low metastatic) and Km12L4A (highly metastatic) (Morikawa et al., 1988.  Cancer Res.  48:1943-1948).  
     [0120] RNA was prepared from each cell line and reverse transcribed to form cDNA. The cDNA was amplified using random primers. Amplification products were visualized on a sequencing gel, and cDNA corresponding to mRNA which was differentially expressed in the cell lines was identified.  
     [0121] Expression patterns and sequence identification numbers of novel metastatic marker polynucleotides are shown in Table 1.  
     [0122] Expression patterns and sequence identification numbers of metastatic marker polynucleotides which correspond to known genes are shown in Table 2. and the corresponding proteins are described below.  
     [0123] Osteopontin (SEQ ID NO:64) (OPN or Spp1 for secreted phosphoprotein 1) is a secreted extracellular matrix protein, often expressed during wound healing, involved in osteoclastic differentiation and activation, as described in Heymann et al., 1998,  Cytokine  10:155-168. Osteopontin is found in bone and other epithelial cells, and has been shown to stimulate proliferation of a quiescent subpopulation of prostate epithelial cells (see Elgavish et al., 1998.  Prostate  35:83-94).  
     [0124] Osteopontin is implicated during the development of diabetic nephropathy (Fischer et al., 1998,  Diabetes  47:1512-1518); the process of cartilage-to-bone transition during rigid bone healing after bone fracture (Nakase et al., 1998,  Acta Histochem  100:287-295); wound healing by an interaction with the receptor integrin alpha(v)beta 3 after focal stroke (Ellison et al., 1998,  Stroke  29:1698-1706); integrin receptor binding and signaling during cell attachment and mechanical stimulation of osteoblasts (Carvalho et al., 1998.  J. Cell Biochem  70:376-390): kidney morphogenesis (Denda et al., 1998.  Mol. Biol. Cell  9:1425-1435); and as an interstitial chemoattractant in renal inflammation (Rovin and Phan. 1998.  Am. J. Kidney Dis.  31:1065-1084). Mice lacking the osteopontin gene showed modulation in osteoclast differentiation from wild type mice (see Rittling et al., 1998  J. Bone Miner Res.  13:1 1011-1111).  
     [0125] Osteopontin is synthesized by monocytes and macrophages within injury sites, and can promote leukocyte adhesion through the alpha 4betal integrin, as described in Bayless et al., 1998.  J. Cell Sci.  111:1165-1174. Osteopontin is transcriptionally regulated by retinoic acid (see Manji et al., 1998,  J. Cell Physiol.  176:1-9); preferentially expressed in high orade metastatic brain tumors compared to low grade brain tumors, and inducible by tissue plasminogen activator (tPA) in glioma cell lines (see Tucker et al., 1998,  Anticancer Res.  18:807-812). Osieopontin is expressed in about 73% of primary gastric carcinoma tissues and corrected with the progression of human gastric carcinoma and lymphogenous metastasis (see Uc et al., 1998.  Int. J. Cancer  79:127-132).  
     [0126] Nip (SEQ ID NO:65) is described in Boyd et al., 1994.  Cell  79:341-351 Adenovirus E1B 19 kDa protein protects against cell death induced by viral infection and external stimuli, and can be functionally substituted with the Bc1-2 protoncogene. E1B 19 kDa interacting proteins (Nip1, Nip2, and Nip3) were discovered in yeast two-hybrid studies conducted to discern proteins that interact with 19 kDa protein, as described by Boyd et al., slipra. Nip 1, 2, and 3 interact with discrete domains of E1B 19 kDa, and similarly also interact with Bc1-2. in both cases promoting cell survival.  
     [0127] Ca −2 -dependent protease (SEQ ID NO:66) is Ca −2 -dependent protease (also called calpain), activity of which is present in every vertebrate cell that has been examined. Ca −2 -dependent protease activity is associated with cleavages that alter regulation of various enzyme activities, with remodeling or disassembly of the cell cytoskeleton, and with cleavages of hormone receptors (see Goll et al., 1992,  Bioessays  14(8):549-556). Ca −2 -dependent protease activity is regulated by binding of Ca −2  to specific sites on the calpain molecule, with binding to each site generating a specific response correlated with a specific activity (e.g., proteolytic activity, calpastatin binding, etc.), as described in Goll et al. Excessive activation of the Ca −2 -dependent protease calpain may play a role in the pathology of disorders including cerebral ischemia, cataract, myocardial ischemia, muscular dystrophy, and platelet aggregation. Therapeutic applications include selective Ca −2 -dependent protease inhibition, as described in Wang and Yuen. 1994,  Trends Pharmacol. Sci.  15(11):412-419.  
     [0128] IGF-R (insulin-like growth factor receptor) (SEQ ID NO:67) is a transmembrane tyrosine kinase linked to the ras-raf-MAPK(mitogen-activated protein kinase) cascade and required for the cell to progress through the cell cycle (Werner and Roith. 1997,  Crit. Rev. Oncog  8(1):71-92). IGF-R mediates mitogenesis, growth hormone action, cell survival and transformation to and maintenance of the malignant phenotype. IGF-R is a member of the growth factor receptor tyrosine kinase superfamily, exists as covalent cross-linked dimers where each monomer is composed of two subunits, and is bound by licand in the extracellular domain (McInnes and Sykes, 1997.  Biopolymers  43(5 ):339-366).  
     [0129] The domains of the IGF-R are described in Sepp-Loienzino. 1998,  Breast Cancer Res Treat  47(3):235-253, including domains responsible for mitogenesis, transformation, and protection from apoptosis. IGF-R expression is increased in breast cancer cells derived from tumor tissue and cell lines, as described in Surmacz et al., 1998,  Breast Cancer Res Treat  47(3):255-267, and increased IGF-R may increase tumor mass and/or aid tumor recurrence by promoting proliferation, cell survival, and cell-cell interactions. Human pancreatic cancers overexpress IGF-R and its ligand (Korc, 1998.  Surg Oncol Clin N Am  7(1): 25-41), and expression of IGF-I and IGF-R is determined to be a prognostic factor (reflecting the interaction between the neoplastic cells and their microenvironment) for lymphocytic infiltration in thryoid carcinomas (Fonseca et al., 1997,  Verh Dtsch Ges Pathol  81:82-96).  
     [0130] ILGF-BP5 (SEQ ID NO:68) is insulin-like growth factor binding protein 5, described in Allander et al., 1994.  J. Biol. Chem.  269:10891-10898. The gene and promoter for IGF-BP5 are characterized by Allander et al., 1994.  J. Biol Chem.  269 : 10891   -10898. and some general actions of IGF-BPs are described in Chan and Spencer. 1997,  Endocrine  7:95-97. Potential impact of IGF-BPs on cancer cell growth is described in Oh, 1997,  Endocrine  7:111-113, and Oh. 1998,  Breast Cancer Res Treat  47:283-293. IGF-BP5 is expressed during brain development: IGF-BP5 and IGF-I mRNAs are synchronously coexpressed in principal neurons of sensory relay systems, including the olfactory bulb, medial and dorsal lateral geniculate bodies, and ventral tier, cochlear, lemniscal, and vestibular nuclei, and are transiently coexpressed in principal neurons of the anterodorsal nucleus, as described in Bondy and Lee, 1993,  J. Neurosci  13(12):5092-5104. IGF-BP5 is expressed by luminal or cumulus granulosa to cells in virtually all follicles, and is highly abundant in stromal interstitial cells of the mature ovary (see Zhou and Bondy. 1993,  Biol Reprod  48:467-482). IGF-BP5 induction is strongly stimulated during differentiation of skeletal myoblasts and is correlated with IGF-R activation as described in Rousse et al., 1998.  Enclocrinolog,  139:1487-1493. IGF-BP5 and other components of the IGF system are critical in postnatal brain development (see Lee et al., 1996.  J. Cereb Blood Flow Metab  16:227-236).  
     [0131] IGF-BP5 stimulates bone cell proliferation by an ICF-independent mechanism involving IGF-BP5-specific cell surface binding sites, as described in Mohan et al., 1995,  J. Biol Chem  270:20424-20431. In connective tissue cell types. IGF-BP5 has a lowered binding affinity to the extracellular matrix which allows IGF-I to better equilibrate with the receptors which in turn potentiates IGF-I action on fibroblasts and smooth muscle cells (Clemmons,  Mol Cell Endocrinology  140:19-24).  
     [0132] Lactate dehydrogenase (SEQ ID NO:69) is a member of the LDH group of tetrameric enzymes with five isoforms composed of combinations of two subunits. LDH-A and LDH-B. Shim et al., 1997,  Proc. Nat&#39;l Acad. Sci.  94:6658-6663. described the relationship between LDH-A and neoplasia. In particular, overexpression on LDH-A may contribute to altered metabolism that confers neoplastic growth advantage. The expression pattern of LDH in the present invention is consistent, in that LDH expression is higher in two metastatic breast cancer cell lines than in a non-metastatic breast cancer cell line (Table 2). High or increasing lactate dehydrogenase (LDH) levels in tumor tissue and cells is associated with poor survival rate in small cell lung carcinoma (SCLC), as described in Ray et al., 1998.  Cancer Detect Prev  22:293-304, making it a useful prognostic indicator for SCLC as discussed in Stokkel et al., 1998,  J. Cancer Res Clin Oncol  124:215-219.  
     [0133] Ufo TKR (SEQ ID NO: 70) is described in Schulz et al., 1993,  Oncogene  8:509-513. This protein has been reported as a marker in tumors, but has not previously been reported in breast cancer. According to the present invention, expression is found in the MDA-MB-231 breast cancer cell line, but not in the MSF-7 or MDA-MB-435 cell lines. This gene and protein provide new markers for distinguishing breast cancer tissue of different types of metastatic potential.  
     [0134] Initially isolated from primary human myeloid leukemia cells, the Ufo oncogene (also called Ax1 or Ark) is a receptor tyrosine kinase (RTK). Its genomic structure is described in Schulz et al, supra, and its differential expression is described in Challier et al., 1996.  Leitkemia  10:781-787. The ufo protein is a member of a class of RTKs having two libronectin type III domains and two imniunoglobulin-like domains present in the extracellular portion, and is preferentially expressed in monocytes, stromal cells, and some CD34-positive progenitor cells (Neubauer et a/l 1997,  Leuk Limphorci  25:91-96). Ufo has an extracellular structure similar to neural cell adhesion molecules, and has direct or indirect binding sites for program GRB2. c-src. and lck (Braunger et al., 1997.  Oncocene  14:2619-2631).  
     [0135] eIF-2 (SEQ ID NO:71) is a translation initiation factor, and functions as a heterotrimeric GTP-binding protein involved in the recruitment of methionyl-tiNA to the 40 S ribosomal subunit (Gasper et al., 1994.  J. Biol. Chem.  269:3415-3422) According to the present invention, higher expression is found in two metastatic breast cancer cell lines and not in cell line MCF-7.  
     [0136] eIF-2 is involved in introducing the initiator tRNA into the translation mechanism and performing the first step in the peptide chain elongation cycle. eIF-2 is associated with a 5 subunit molecule having GTP recycling function called eIF-2B (Kyrpides and Woese. 1998,  Proc. Nat&#39;l Acad. Sci.  USA 95:3726-3730, and Kimball et al., 1998,  J. Biol. Chem.  273:12841-12845).  
     [0137] eIF-2 has subunits alpha and beta. eIF-2alpha is phosphorylated at Ser 51 and then modulates the interaction of eIF-2 and eIF-2B, as described in Kimball et al., 1998.  Protein Expr. Purif.  12:415-419, Kimball et al., 1998,  J. Biol. Chem.  273:3039-3044, and Pavitt 1998.  Genes Dev.  12:514-526. It is reported that by regulating translation initiation, control of cell growth and division in eukaryotic cells is achieved: for example, clotrimazole, a potent anti-proliferative agent in vitro and in vivo, depletes intracellular Ca −2  stores, which activates PKR, resulting in the phosphorylation of eIF-2alpha, and the ultimate inhibition of protein synthesis and blockage of the cell cycle in GI phase (Aktas et al., 1998,  Proc. Nat&#39;l Acad. Sci.  USA 95:8280-8285). Additionally, Kim et al., 1998.  Mol. Med.  4:179-190. show that nitric oxide (NO) suppresses protein synthesis in cell types including human ovarian tumor cells by stimulating phosphorylation of eIF-2alpha.  
     [0138] Glutaminyl cyclase (SEQ ID NO:72) is described by Song et al., 1994.  J. Mol. Endocrinol.  13:77-86. and is expressed most highly in the most metastatic cell line MDA.-MB-435. as compared to less metastatic line MDA-MB-231 and non-metastatic line MCF-7. Glutaminy 1 cyclase (also called lutamine cyclotransferase) converts zlutaminyl-peptides (such as gonadotropin-releasing hormone and thyrotropin-releasing hormone) into pyroglutamyl-Deptides, as described in Busby et al. 1987.  J. Biol. Chem.  262:8532-8536. Fischer and Spiess, 1987.  Proc. Nat&#39;l Acad. Sci.  USA 84:3628-3632, and Pohl et al. 1991.  Proc. Nat&#39;l Acad. Sci.  88:10059-10063. Cloning and sequence analysis of glutaminyl cyclase derived from a human pituitary cDNA library is described in Song et al., 1994.  J. Mol. Encocrinol.  13:77-86Studies on the catalytic pathway of glutaminyl cyclase and its substrate specificity are described in Gololobov et al., 1996.  Biol. Chem. Hoppe Seyler  371:395-398. Assays for the presence of glutaminyl base activity are described in Koger et al., 1989,  Method Enzymol.  168:358-365 and Houseknecht et al., 1998.  Biotechniquiies  24:346.  
     [0139] gp130 (SEQ ID NO:73) is transmembrane protein glycoprotein 130. gp130 is a signal transducing shared component of the receptor complexes for the interleukin-6 (IL-6)-type cytokines (Hirano et al., 1997,  Cytokine Growth Factor Rev.  8:241-252). including IL-6, IL-11. leukemia inhibitor factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor and cardiotrophin-1. The N-terminal of gp130 is an extracellular immunoglobulin-like portion of the protein (Hammacher et al., 1998.  J. Biol. Chem.  273:22701-22707). Signal transduction including gp130 occurs through the gp130/Jak/STAT pathway 1 (Heinrich 1998,  Biochem. J.  334:297-314). The cytokines acting through the pathway that includes gp130 (also called gp130 cytokines) exhibit pleitropic biological activities including immune, hematopoietic, and neural effects (Nakashima and Taga, 1998,  Semin Hematol.  35:210-221. Thompson et al., 1998.  Neuroscience  84:1247-1255, Hirano, 1998.  Int. Rev. Immunol.  16:249-284, Marz et al., 1997. Eur. J. Neurosci. 9:2765-2773, and Betz and Muller, 1998,  Int Immunol  10:1175-1184).  
     [0140] gp130 cytokines are reported to control survival and proliferation of myeloma cell lines and primary myeloma cells (Klein. 1998.  Curr. Opin. Hematol.  5:186-191). gp130 is expressed in the majority of renal cell carcinomas and has an important role in the proliferation of some renal cell carcinoma cell lines (Costes et al., 1997,  J. Clin. Pathol.  50:835-840).  
     [0141] E-cadherin (SEQ ID NO:75) is a member of a family of glycoproteins responsible for calcium-dependent cell-cell adhesion and is implicated in maintaining cytoskeletal integrity. Epithelial cadherin (E-cadheriti) mediated cell adhesion system in cancer cells is inactivated by multiple mechanisms corresponding to the pathological features of the particular tumor type (Hirohashi. 1998,  Am J Pathol  153:333-339). In cgeneral the cadherin system mediates Ca −2 -dependent homophilic cell-cell adhesion. Transcriptional inactivation of E-cadherin expression occurs frequently in tumor progression, and thus inactivation or downregulation of E-cadherin plays a significant role in multistage carcinogenesis (Hirohashi. 1998.  Am J Pathol  153:333-339).  
     [0142] E-cadherin is characterized as a tumor suppressor of the metastatic phenotype, as described in Macrogan and Bookstein, 1997.  Semin Cancer Biol  8:11-19, and cadherins are important determinants of tissue morphology including invasive carcinoma as described in van der Linden. 1996,  Early Pregnancy  2:5-14. and Yap. 1998,  Cancer Invest.  16:252-261.  
     [0143] Mechanisms of action of cadherins are discussed in Daniel and Reynolds, 1997.  Bioessays  19:883-891. The structure and function of cell adhesion molecules including E-cadherin are described in Joseph-Silverstein and Silverstein. 1998,  Cancer Invest.  16:176-182, Yap et al., 1997,  Annu. Rev. Cell Dev. Biol.  13:119-146, and Uemura, 1998.  Cell  93:1095-1098. Cell adhesion molecules including E-cadherin are potential targets for anti-cancer drugs and therapeutics to treat acute or chronic inflammatory disease as described in Buckley and Simmons. 1997.  Mol Med Today  3:449-456, Moll and Moll. 1998.  Virchows Arch  432:487-504.  
     [0144] According to the present invention. E-cadherin is expressed in non-metastatic breast cancer cell line MCF-7, and not in MDA-MB-231 and MDA-MB-435. The expression products are diagnostic markers indicating the metastatic potential of breast cancer tissue samples.  
     [0145] Serpin (SEQ ID NO:76). serine protease inhibitors, are a family of protease inhibitors that inhibit chymotrypsin-like serine proceases (Whisstock et al, 1998.  Trends Biochem. Sci.  23:63-67) and that have the unique ability to regulate their activity by changing the confirmation of their reactive-center loop: studies, of serpin variants provide definition for the functional domains of serpins that control the folding and link serpins mutations to disease (see Stein and Carrell. 1995.  Nat. Struct. Biol.  2:96-113) Serine protease cleavage of proteins is essential to a wide variety of 20 biological processes, and the cleavage is primarily regulated by the cleavage inhibitors, as described in Wright, 1996.  Bioessays  18:453-464. Members of the serpin family include alpha I-antitrypsin (AAT) (Carrell et al., 1996,  Chest  110:243S-247S), alpha2-anti-plasmin (PAI-1 and PAI-2) (Andreasen et al., 1997.  Int. J. Cancer  79:1-22), thrombin, urokinase plasminogen activator, and kallikrein (Turgeon and Houenou. 1997.  Brain Res Brain Res Rev  25:85-95). Some serpins also have other activities including neuronal differentiating and survival activities (Becerra, 1997.  Adv. Exp. Med. Biol.  425:332-237) and tumor suppression (Sager et al., 1997.  Adv. Exp. Med. Biol.  425:77-88). PAI-1 and PAI-2 are linked to cancer metastasis, as described in Andreasen et al., 1997.  Int. J. Cancer  72-22.  
     [0146] pS2 (SEQ ID NO:77) was isolated from MCF7 human breast cancer cells, as described in Takahashi et al., 1990,  FEBS Letters  261:283-286. pS2 is estrogen-regulated. Speiser et al., 1997,  Anticancer Research  17:679-684, reported that the pS2 status declined from well to poorly differentiated ovarian cancer. pS2 expression also is associated with a good prognosis in breast cancer patients. According to the present invention, pS2 is expressed in MCF-7 cells, but not in two metastatic breast cancer cell lines  
     [0147] pS2 (presenilin-2 or trefoil factor 1 (TFF 1)) is a trefoil polypeptide normally expressed in the mucosa of the gastrointestinal tract, and found ectopicallv in gastrointestinal inflammatory disorders and various carcinomas (May and Westlev. 1997,  J. Pathol.  183:4-7. pS2 is expressed in breast cancers (Poulsom et al., 1997.  J. Pathol.  183:30-38), pS2 is a pleitropic factor involved in mucin polymerization, cell motility (Modlin and Poulsom. 1997,  J. Clin. Gastroenterol  25(j):S94-S100). cell proliferation and/or differentiation, and possibly in the nervous system (see Ribierms et al., 1998,  Biochim. Biophys. Acta. ! 378:F61-F77).  
     [0148] LIV-1 (SEQ ID NO:78) is an estrogen-regulated protein reported in the MCF-7 cell line (Green et al., GeneBank submission Accession No. U41060) According to the present invention, LIV-1 is expressed in MCF-7 cells, but not in to metastatic breast cancer cell lines.  
     [0149] Leucine-isoleucine-valine-1 (LIV-1) and other members of the LIV family (LIV-2, 3, and 4) are binding proteins that represent a transport system for branched chain amino acids in  E coli  as described in Yamamoto et al., 1979.  J. Bacterial.  138:24-32. and Yamamoto and Anraku. 1980.  J. Baicteriol.  144:36-44. A human homologue to LIV-1 is both estrogen and growth factor inducible in MCF-7 human breast cancer cell line (El-Tanani and Green, 1997.  J. Steroid. Biochem. Mol. Biol  60:269-276; El-Tanani and Green. 1996.  Mol Cell Endocrinol  124:71-77: and El-Tanani and Green, 1996,  Mol Cell Endocrinol  121:29-35).  
     [0150] GTP-binding protein (SEQ ID NO:79) is a member of the family of guanine nucleotide-binding regulatory proteins. G proteins. The protein is expressed in MCF-7 cells, but not in two metastatic breast cancer cell lines.  
     [0151] G proteins provide signaling mechanisms for the serpentine family of receptors as described in Dhanasekaran and Prasad, 1998,  Biol. Signals Recept  7:109-117. Studies indicate that the alpha as well as the beta gamma subunits of the GTP-binding proteins are involved in the regulation of several cellular responses, some of which responses are critical to the regulation of cell growth and differentiation (Dhanasekaran and Prasad. 1998,  Biol Signals Recept  7:109-117). G protein coupled receptors regulate the mitogen activated protein kinase pathway as described in Russell and Hoeffler, 1996,  J. Invest. Dermatol Symp Proc  1:119-122, and thus play a role in controlling cell growth. GTP binding proteins are also implicated in the regulation of intracellular transport as described in Ktistakis, 1998,  Bioessays  20:495-504.  
     [0152] Chemokines induce various intracellular signaling pathways in natural killer cells by activating members of GTP binding proteins as described in Maghazachi and Al-Auokaty. 1998.  FASEB J.  12:913-924 Heterotrimeric GTP binding proteins regulate distinct signaling pathways, some of which in turn regulate the activity of Na+/H+ exchanger proteins as described in Voyno-Yasenetskaya, 1998,  Biol Signals Recept  7:1 18-124.  
     [0153] Desmoplakin (SEQ ID NO:84) is a member of a family of proteins that serve as cell surface attachment sites for cytophasmic intermediate filaments.  
     [0154] Vimentin (SEQ ID NO: 80) is a member of the intermediate filament gene family (Evans. 1998.  Bioessays  20:79-86. Intermediate filaments are a major component of the cytoskeleton of higher eukaryotes. Vimentin gene knockout mice indicate degeneration of the cerebellar Purkinje cells (Galou et al., 1997.  Biol Cell  89:85-97). Vimentin is positive in immunohistochemical reactions of sarcomas and related lesions (Gaudin et al., 1998,  Am J Surg Pathol  22:148-162), and of desmoplastic small round-cell tumors and their variants (Gerald et al., 1998,  J. Clin. Oncol.  16:3028-3036). Vimentin is also expressed in neoplasms showing follicular dendritic cell differentiation as described in Perez-Ordonez and Rosai. 1998.  Semin. Diagn. Pathol.  15:144-154. and in biphasic carcinomatous-sarcomatous malignant mixed mullerian tumors as described in Guarino et al., 1998.  Tumori  84:391-397.  
     [0155] Cytochrome C Oxidase (CcO) (SEQ ID NO: 81) is the terminal enzyme of the respiratory chain of mitochondria and aerobic bacteria: it catalyzes electron transfer from cytochrome C to molecular oxygen, reducing the oxygen to water (Michel et al., 1998,  Annu Rev Biophys Biomol Struct  27:329-356). Cytochrome C oxidase is a member of the superfamily of quinol and cytochrome C oxidase complexes that are related by a homologous subunit containing six positionally conserved histidines that ligate a low-spin heme and a heme -copper dioxygen activating and reduction center as described in Musser and Chan, 1998.  J. Mol. Evol.  46.508-520. Cytochrome C and ubiquinol oxidases are membrane-bound redox-driven proton pumps which couple an electron current to a proton current across the membrane (see Karpefors et al., 1998,  Biochim Biophys Acta  1365:159-169). Analysis of mutant forms of cytochrome C oxidase is described in Mills and Ferguson-Miller. 1998.  Biochim Biophys Acta  365:46-52. Nitric oxide inhibits respiration at cytochrome C oxidase, as described in Torres et al., 1998.  J. Bioenerg Biomembr  30:63-69.  
     [0156] Heat shock protein 90 (hsp90) (SEQ ID NO: 82) acts as a chaperone molecule in association with the glucocorticoid and progesterone nuclear receptors, and has A, B, and Z regions for facilitating these interactions (Dao-Phan et al., 1997.  Mol Eniocrinol  11:962-972). Levels of hsp90 are reported elevated in active systemic lupus erythematosus (Stephanou et al., 1997.  Biochem J  321-103-106). Increased hsp90 expression is implicated in regulation of forms of cell injury that lead to programmed cell death as described in Galea-Lauri et al., 1996.  J. Immunol.  157:4109-4118. Hsp90 is upregulated in regenerating fibers and diseased fibers of Duchenne muscular dystrophy (Bonnman et al., 1996.  Muscle Verve  19.574-580), and is a candidate substrate for proteolysis during ionizing radiation-induced apoptosis of some breast cancer cells (Prasad et al., 1998,  Int. J. Oncol  13:757-764). Hsp90 is involved in dislocation of the mutant insulin receptors from the endoplasmic reticulum to the cytosol as described in Imamura et al., 1998.  J. Biol. Chem.  273:11183-11188. and associates with and activates endothelial nitric oxide syntheses as described in Garcia-Cardena et al., 1998,  Nature  392:821-824.  
     [0157] Inteorin alpha 6 (SEQ ID NO: 83) is in the family of integrins, heterodimeric, cation dependent cell membrane adhesion molecules that mediate cell-cell and cell-matrix interactions. Integrin alpha 6 is a component of the hemidesmosome complex (Jones et al., 1998,  Bioessays  20:488-494). Integrins maintain tissue integrity and regulate cell proliferation, growth, differentiation, and migration. (See Thomas et al., 1997,  Oral Oncol  33:381-388). In oral squamous cell carcinomas there is a variable loss or reduced expression of integrin alpha 6. as described in Thomas et al., 1997,  Oral Oncol.  33:381-388. Alpha 6 integrin also plays an active role in invasion of intestinal and diffuse-type cells of representative gastric carcinoma cell lines as described in Koike et al., 1997.  J. Cancer. Res. Clin. Oncol.  123L:310-316.  
     [0158] Osteozenic crotein-1 (OP-I) (also called BMP-7) (SEQ ID NO: 85) is a morphogenetic factor (and a member of the bone morphogenetic protein (BlvIP) family of growth factors) and is highly expressed in kidney and involved in tissue repair and development (see Almanzar et al., 1998.  J. Am Soc. Nephrol.  9:1456-1463). OP-1 is also expressed in the developing nervous system and can induce dendritic Irovth in sympathetic neurons as described in Gut et al. 1998.  Neurosci. Lett  245:131-134. OP-1 stimulates cartilage formation as described in Klein-Nulend et al. 1998.  J. Biomed. Matter. Res.  40:614-620.  
     [0159] OP-1 induces down-regulation of insulin-like growth factor binding proteins (particularly IGFBP-5) thus affecting IGF-1 in the context of bone cell differentiation and mineralized bone nodule formation as described in Yeh et al., 1997,  Endocrinology  138:4181-4190. OP-1 can be used as a bone graft substitute to promote spinal fusion and to aid in the incorporation of metal implants (Cook and Rueger. 1996.  Clin. Orthop.  324:29-38). The three dimensional structure of OP-1 is reported in Griffith et al., 1996.  Proc Nat&#39;l Acad Sci  93:878-883.  
     [0160] The protein encoded by SEQ ID NO:56 is a putative secreted protein and is highly expressed in fat tissue.  
               TABLE 1                          Novel Differentially Expressed Metastatic Marker Polynucleotides                                                     breast                               cancer   breast                   metastatic   cancer   low               non-   to bone   meta-   meta-               meta-   and/or   static   static   high       TRAN-   SEQ   static   lung   to lung   from   metastatic       SCRIPT   ID   breast   MDA-   MDA-   colon   from colon       NUMBER   NO:   MCF-7   MB-231   MB-435   KM12C   KM12L4A                                                 901   1   −   +   −               907   2   −   −   +       9102b   3   +   −   −       9114   4   −   −   +       9121a   5   −   +   −       9129   6   +   −   +       9139a   7   +   −   −       9143b   8   +   −       9157b   9   −   −   +       9166   10   +   −   −       9170b   11   −   +   −       9190a   12   +   −   −       9191   13   −   −   +       9216   14   −   −   +       9224c   15   +   −   −       9230b   16   +   −   −       924   17   +   −   −       9242a   18   −   +   −       9259a   19   −   −   +       9261   20   −   +   −       9272   21   +   −   −       9293b   22   −   +   −       9304b   23   +   −   −       9307a   24   −   +   −       931   25   +   −   −       9313   26   −   −   +       9316   27   +   +   −       9318b   28   +   −   −       9320a   29   −   −   +       9330b   30   −   +   −       9335   31   +   −   +       9337   32   +   −   +       9342b   33   −   +   −       9343c   34   +   −   −       9350e   35   −   +   −       9351b   36   −   +   −       9361   37   +   −   −       9368   38   −   +   −       9373b   39   −   −   +       9385a   40   −   −   +       9386c   41   −   −   +       9388d   42   +   −   −       9390   43   +   −   −       9393   44   +   −   −       9396   45   −   +   −       944b   46   +   −   −       951   47   +   −   −       953   48   −   −   +       954a   49   +   −   −       968   50   +   −   −       971   51   +   −   −       983c   52   −   +   −       985   53   +   −   −       990   54   +   −   +       998   55   −   −   +       316   56   +   −   −   +   −       126c   57   −   −   +       207-4   58   −   +   −       265-3   59   +   −   −       29B   60   −   −   +       305B-25   61   +   −   −       326B-39   62   +   −       34B-11   63   −   −   +                                  
 
     [0161] For transcript number 316, reverse transcription PCR (RT-PCR) was used to detect expression in the breast cancer cell lines.  
               TABLE 2                          Differentially Expressed Metastatic Marker Polynucleotides                                                     breast                           cancer   breast                       metastatic   cancer                   non-   to bone   metastatic       TRAN-       SEQ   metastatic   and/or lung   to lung       SCRIPT       ID   breast   MDA-MB-   MDA-       NUMBER   protein   NO:   MCF-7   231   MB-435               902   osteopontin   64   −   −   +       9112   nip   65   −   +   −       9132   Ca-dependent   66   −   +   −           protease       9158   IGF-R   67   +   −   −       9174   ILGF-BPS   68   +   −   −       9177   lactate   69   −   +   +           dehydrogenase       9202   ufo TKR   70   −   +   −       9210   eIF2   71   −   +       9212   glutaminyl   72   −   −   +           cyclase       9213   gp130   73   −   −   +       9222   TGFb-II   74   −   +   −       9232   E-cadherin   75   +   −   −       9239   serpin   76   −   +   −       9250   secreted pS2   77   +   −   −       9260   LIV-I   78   +   −   −       9315   GTP-binding   79   +   −   −           protein       9317   vimentin   80   −   +   −       938   cytochrome C   81   +   −   −           oxidase       9382   Hsp 90   82   −   −   +       9394   integrin a6   83   −   −   +       956   desmoplakin   84   +   −   −       970   osteogenic   85   +   −   −           protein                                  
 
     [0162] Within the scope of the invention are variants of the proteins described above. A variant is a protein encoded by a polynucleotide wherein the global sequence identity of the DNA, as compared to the corresponding SEQ ID NO: herein, is at least 65% as determined by the Smith-Waterman homology search algorithm as implemented in MPSRCH program (Oxford Molecular) using an affine gap search with the following search parameters: gap open penalty of 12, and gap extension penalty of 1. The protein encoded by the DNA having the sequence identity described above will exhibit the percent activity described in the preceding paragraph.  
     [0163] Also within the scope of the invention are fusion proteins comprising the proteins and variants disclosed herein. Proteins preferably used in fusion protein construction include beta-galactosidase, beta-glucuronidase, green fluorescent protein (GFP), autofluorescent proteins including blue fluorescent protein (BFP), glutathione-S-transferase (GST), luciferase, horse radish peroxidase (HRP) and chloramphenicol acetyltransferase (CAT) Additionally, epitope tags are used in fusion protein constructions, including Histamine (His) tags. FLAG tags, influenza hemagglutinin (HA) tags, Myc tags, VSV-G tags, and thioredoxin (Trx) tags, other fusion constructions can include maltose binding protein (MBP), S-tag, Lex A DNA binding domain (DBD) fusions, GAIL4 DNA binding domain fusions, and Herpes simplex virus (HSV) BP16 protein fusions.  
     [0164] These fusions can be made by standard procedures in the art of molecular biology, and many are available as kits from, for example. Promega Corporation (Madison. Wis.); Stratagene (La Jolla, Calif.); Clontech (Mountainview, Calif.); Santa Cruz Biotechnology (Santa Cruz. Calif.); MBL International Corporation (MIC, Watertown, Mass.); and Quantum Biotechnologies (Montreal, Canada).  
     [0165] The proteins of the invention, and variants as described herein, can also be used to detect protein interactions in vivo, using the yeast two-hybrid system, for example as described in U.S. Pat. No. 5,674,739.  
     [0166] In addition to the ribozyme and antisense constructs described above, the polynucleotides of the invention can be used for inhibiting transcription via triple helix formation as disclosed in U.S. Pat. No. 5,674,739.  
     [0167] Those skilled in the art will recognize, or be able to ascertain, using not more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such specific embodiments and equivalents are intended to be encompassed by the following claims.  
     [0168] All patents, published patent applications, and publications cited herein are incorporated by reference as if set forth fully herein.  
    
     
       
         1 
         
           
             85  
           
           
             1  
             142  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(142)  
               n = A,T,C or G  
             
           
            1 

cacccaagaa ctaagaaaca aagggagaat gtacttttgt agcttagata agcaatgaat     60 

cagtaaagga ctgatctact tgctccacca cccctccctt aataataaca tttactgtnn    120 

atttcctggg cctaagactc ta                                             142 

 
           
             2  
             331  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(331)  
               n = A,T,C or G  
             
           
            2 

cgcgagcaga caacataatt tatttccaga aaacaacaga aatgaacatc atcatgaata     60 

catgaaatcg gctgtgatgt gtgaactgct aagggccaaa tgaacgtttg cagagcagtg    120 

ggcacaatgt ttacaatgta tgtgtatgtc actttcggta cctgtgaatg catggggacg    180 

tgctgaaccc gaaaaaaagt gcctttccat aaggactgca atanagaggg caatttaccc    240 

tggtggtaca cggaacctan attcactcct gccatgcctt gccaatagta anctgcaggg    300 

tggaacaaga aatcacttgc tctgggggga a                                   331 

 
           
             3  
             1112  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(1112)  
               n = A,T,C or G  
             
           
            3 

ccnnnnnnnn ntncntnnnn ncnnnnccnn ngnnnnnctn gcccnnncng ctnnnccccn     60 

nntnctnntn gntnangnnc ngaanccgcn nnnngnnnnn acnatnntnn gncgnnnnnt    120 

tcgttnnnnc ntgnntccnc nnnnnctngt ncnnnnnggn ggngcgcncc nccnancctn    180 

cctcnntgnn ncnnnctnnt nnctnngctg ngtctcncng cncngngcnn nnnggggtct    240 

nccgtnctnc nnnnncnnng ttttangncn gnaanacgcc gcgncgagct tttagccatg    300 

ggggataacc gaaccaaacn tnacactctc agaggatcca cctntgggtg caagcgaaac    360 

tngancnatc tatactctcg anggtncaag gacattgntg agagaaatgg anncacagcc    420 

cacgttcatt gggtangaga ctccnattaa natttctgtc tccccngatg ggccctagac    480 

ccatgaatcc ctattangat cccntcagcg gccanacncn gtggctccnc ctgtaatccc    540 

ccacntcggg aggctgatga gggcgaatcc aaggtcagga aatntatata gacncctggc    600 

taaccggnga accccccctc taaaancaaa aaaaaanncc nncnngtntt tanagggngt    660 

tntttttcnt cgccncgccc gncncgnccg cttnctngct ccncctgnnc nnncntccct    720 

ncnncnntgn tcancccngc gnnncgcnnc ntnccttnnt gngtctggtc ncncttcnnc    780 

ctctcttncn ccnntgtccn tngctctcag ccnctgcccc nccctnnccn tnngtgnnnc    840 

cnccntnatg nccncncnan aggngcangc nntggcncgc tgnccnntgt ntgtcnctcn    900 

acgganantg nactcncnac tnngnnacgc natnnnanct ctgctctcag atgacagcan    960 

cggnntnnnc ngcctctanc nncngnnncn nagccnncga nnnaggnanc cgcgntcant   1020 

cnnntttcnc tctncnntng catntctgat ngccgtgnct ncctcnnttn ctcnagcncn   1080 

tnnccacctc tcgtttagnc nctnnncnna nn                                 1112 

 
           
             4  
             183  
             DNA  
             Homo sapien  
           
            4 

aaaactatga attccatact tgaggtttcc cagccaattg ctcccttctg ctttagaagt     60 

gactaggtac tgagagtaca aacactccca ctttataatg aaggcgtcat gtcacccctt    120 

cctttacagg tcctggggtc caggagaccc agaatgaagg tgtcagttgg gcatgaagtg    180 

tta                                                                  183 

 
           
             5  
             1092  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(1092)  
               n = A,T,C or G  
             
           
            5 

ttncagacca agaagacttg atnagctgaa acccattgcn ctacttggaa ngtgatcngc     60 

aaaagctgcc tcagtcanac accggggata aatctggatt tgggttccgg cgtcaaggtg    120 

aanatnatac ctantaanga acnctgtaca ntgccncaag cangtganga ccncccacga    180 

gtttacatna atacaatnct gaaacnacnc aggctggttt tatatctaca tatttgactt    240 

accactatcn cantaaagtt tngcaccttt cnccgaacga aaanaacccc ccntnntgnn    300 

ttcttttnaa aanaccntng nnccncnttn ccgtcncncc ccnnatantn nncnnatccc    360 

cccctctncc nntccntnnn cgtaannggc gtngcttntg cngtntntgt cccgttttcc    420 

tccgcttngt cntttntcta tatnggctnn tnttatnccn ngcccttcgt cncctnnngn    480 

ttcgtctgtn cntagtcctc ntnctngagc cccanttgnt acttcnngct tcnnctccgc    540 

attccntctc cgcncnnanc ncnnntctca nannatgnnc nntnnctncn nccnatncnc    600 

cctnanagnt tcgnctagac cntcnacntt gtntcccgnn ctcttagngn tctgctncta    660 

gtgtntnnct catctcctct ncttctctct cctttgacnc ngnncnctcc atcntnntct    720 

gnctttctca tcncnnnnng cccctnctcn cnnagtntgn gtgcncnnnc ttnnnntcna    780 

nctngtcgcc tccgttttcn actnnnnccn nngcngnncg nnngctcttt ctntcnntta    840 

gactnacctt ntctgnnnnn tcannctagc nctgtccntc tctnntctgc atcnttanac    900 

atcttnntcn cccnctcgca ncntnctntt nacnctcnca tacgttnccn nnctcagtcc    960 

gcagnnnngt tncntncngt cntctcgcgn ctcnnntcct ctctnnnacn cncctggtct   1020 

ncgnctcgct ccnncccatn cntncctcgt tgntcnnnnt cnnatacgtn tncangccnc   1080 

ntctctccnc tn                                                       1092 

 
           
             6  
             504  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(504)  
               n = A,T,C or G  
             
           
            6 

ctggagcggg atcatttana atactttaca gatatntgca ccaggtacat ntatntgcgt     60 

ccattggtag cacagctgag acctgtgtct cacatcagcc taggtgaagc ctactacaaa    120 

taatgccaag ggagaanagc cagtacacta tatggtttat actctttatc cctttattca    180 

tagcatgttt tttaaaaatg ttatattatg caacagatgt gaggcagcan ctaagctata    240 

cttaagaatt ttctctcacc ttccaaacca aagtgtcctg aataagccag gagacttatt    300 

cttttgtgca ccctggtgca catctgactg ttgtcctanc canaaactct ctgaggccac    360 

tgaaagaaca gtggccctat cgatttcatt cctaggtctc aaaaatacna tgtngccttg    420 

taacataatt agggacagca cctctatttc acaattataa tctaaggtag gataagacga    480 

cacagcagca ataaacttac aagt                                           504 

 
           
             7  
             1132  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(1132)  
               n = A,T,C or G  
             
           
            7 

gcgngccccc tngtngnncn ttntncncng ttttctgctn tntttatnng aggnctnggt     60 

nnttnntctt agggnnntng tncggtcnng ttnntgttnc gagcagaaag tgnatatttc    120 

atgcngccaa gcttntttat tgaaaantcc taattntatt gnccgtntag taacatgttt    180 

gttcnacaan gctaatttct nataaancaa aacacannnt tttcttataa gtngtataaa    240 

ttatttnatt tacagaaact tgtttcaaaa canatgnact anntatttct nctcttttaa    300 

atanccanac taattttcta tccctngaca tctgttcatg ttctatncag cagccaacac    360 

aaagtccanc tgagagctct tgattaangt gtncgnatta tctagctact tccnacgttt    420 

tnggngcnng aaatgncttt taanancctg gcctcaaaaa anaaaaanan ccccccgnnn    480 

aggggnnttc cntntanaaa aanggntcnc tcnnccngtn ngagactgtc tccctgnntn    540 

ngnnnntcgc tntnatcang ngccncnang ctcnccntcn ctnnngcatt ngatnnntan    600 

cnnnctgaga tgngnntang ctgntncntn ngtgtcntan gtctcgacgt tgnntggntn    660 

tangnancgn cnntntnnnc nnattgncga gngnntaagt gtgctcttct cntnacntct    720 

ntcnnnancn tctnngatgt tnatacggcc gtgcttnctt atcnntgana ncgntctnan    780 

nanntncgna tgagnntnta ctgcncncnt gtgtcatctt tctctctant gtgtnctnna    840 

nncnngtnat tncgcnnnac tgntantnag tggtatnnag anntcgnncg cnngngccnn    900 

tttnnctgtn gnnatnagnt ntcanganat tnatncnntc tncgtgatag anagntnagt    960 

gnnggntctg actgatncgt gtcctagtnn cngtgacatc gnncgttann gtcngcactc   1020 

tagtanannt nagtnngang ntgtanatnn ntctcntgtt tcagtnnagn cccncgagcg   1080 

cntcanntnt nantgtctcn tctnngtcgt anncntgtcg agtngtnana nn           1132 

 
           
             8  
             736  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(736)  
               n = A,T,C or G  
             
           
            8 

ntgggcccga cgtcgcatgc tcccggncgn catggnnnnc tggtttggtc anatgtgaat     60 

aacgnagaan tgagaccacn ganaagaacc acantgtnan ggnncttgca cntgntanga    120 

antnagnaat gcctttttnc tgagggcntt nggnnntcat nnangggngt gnggnggntt    180 

ncacctgtaa taccaccact ttncnatgcc actgccngtg natcaccngn ngtaaggact    240 

tcaanaccag ccttatnaac ntgggnaaac cntntntcta ctaaaaatnc tnnaantatc    300 

tgngcnnngt ngngcgttct tntannnccn gctgnacnng angncngngn angntantcg    360 

cntgaacntg ncntgttana gtngcantga gcctaaatca cantgatgta ttnncatctg    420 

ggacgacacg ancngacgac tcncgtactn aaaaaaaaaa ncccnttnng ggggggtttt    480 

tnnnggtatt anntatantt ggagaanttt gggtcannng aatattntta catgaaaaat    540 

naggaataac tntatntgtg tacattgggt tnnaaanang acantantgg nnctaaactn    600 

ttnggggngg aggggnnatt agggnnttaa ttnggnnnct tnnaaanncn nntnnngtat    660 

nanaanantn tttnnanaag ngnantngnt ttaaancctn aangnttnnn tnctnttann    720 

ttnnaannnn anannn                                                    736 

 
           
             9  
             690  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(690)  
               n = A,T,C or G  
             
           
            9 

tnnncctggn tggtcactcc cttctgtcct gttagctcat ggtgtaagat gatgtcttgt     60 

cagtattact gttttgctaa gccgcttcat tcatgcctac acaatttttt tttaaaaggg    120 

aactttagtt aattaagtga taagggactt aaatatgaat tanaatggtg cagaaagaga    180 

taccttttct ggatatttta aagtttaaag gtcantttct cttaatctga ttatgtgcac    240 

atatgaaaat ggcacatcat atacatgtaa aatcaggcag tatncattta ttaattactg    300 

tatttgacaa aggaaactct taaattataa tgtgaaacct ggttttatga aaccaatgac    360 

tagtgcanca tttcagcata tgcaaaaaaa aaanncctnt tggngngctg tttacaaagg    420 

aaattgttgg atttcacgat ggtttcagga naanaaggtt ttcntcatcn agggtaaacn    480 

tcccggataa ggcntngntt taatntnntt annccnnccn atngntaann gtggaaatta    540 

ancctctgaa naaaanancc cacntnnttn gccttgggct tnantctntt tggcngnanc    600 

naaaggnnct tnccaggtnt cntgnngggc cngnngaann ataannaann nggggnnctt    660 

nggaaacctt ncnnnaanan tncccncccc                                     690 

 
           
             10  
             395  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(395)  
               n = A,T,C or G  
             
           
            10 

tggtatctga cnnaataaga atgcacccat ttgtgagggg taatatttat ctcangattt     60 

actgtaaata tgtatacaca catacaaaaa cccaggcatt gttaagagaa aatnatggcc    120 

cagaggttna aattatcaga cagaaccttt aanaataatt atgattaatg tgttaaaatt    180 

ctagtggaaa agataaataa catgctcagg anattttagc anagagatag aaactatntn    240 

ngaagctcaa atgaaaatgc taggaaatga aaagcagtat tggaggtgaa agattccttt    300 

ggcaatttat caacanactg gagatggcan aggcataatc agtantattg aaggcagatt    360 

actatntatt atncaancaa aaaaaaaaac cccct                               395 

 
           
             11  
             331  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(331)  
               n = A,T,C or G  
             
           
            11 

aacgaggccn ngaggccaat gaggccaaca agacgatgcc ggagacccca actggggact     60 

cagacccgca acctgctcct aaaaaaatga aaacatctga gtcctcgacn atactagtgg    120 

ntcgctacag gagggaacgt gaaaagaaca tctccagagg aactggtgaa tgaccacgcc    180 

cgagagaaca gaatcaaccc cgaccaaatg gaggaggagg aattcataga aataacgact    240 

gaaagaccta aaaagtagca agaagctaca tccctcaaac ttcggcaatg aaaataaagt    300 

ttgagaagct caaaaaaaaa aanccctttt g                                   331 

 
           
             12  
             693  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(693)  
               n = A,T,C or G  
             
           
            12 

tncaacgcgt tgggagctnt nccaaggtgg nctagcnnca ttaatgccct accgtgggaa     60 

tatggntgaa gatcttgact aggggactta tgaacccatg cagccgtgcc caaatcctac    120 

caaactgacc ttactttctt gaagacggaa ttgtagtatg gtcgagctca tgctttttgt    180 

agtaggccat ncaaattcga ttgactggct aaaaaagatt gttagtggag gctggaagaa    240 

acattttggc tgatgataga tgaatagagc ttggaacaat caaaaggaaa agcagaaagt    300 

ctatacctat tcataagaaa aagttagtat gtttaccgaa cattatnaaa gaattatgac    360 

attttcaaag ttttaaaatt ttattttgta gggacggggt ctcattgtgt agcccacnct    420 

ggtctgtttc ttgaggattt actatanact gggctgtatt caaagcattg gggatacagg    480 

catgaatgag cccccattgc ctgaacttac cattcaatct gggcagtgaa agaanaggga    540 

tgntgggaga nccttacaaa gatgaaatgt cgctaactgg agaaatccct actttcagtc    600 

agactgaann ggaacaggta gtnactgtgg gtagccctct ttgggnangg gtngattttc    660 

cacatgtgcc cagttaaggg ccnagaacat taa                                 693 

 
           
             13  
             305  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(305)  
               n = A,T,C or G  
             
           
            13 

ttggtatcng gggatgggng aggggagata gncccgaagc atcccnnatt ctcagtaaac     60 

tccttggnat canannatat cntggccnaa gaaccncnca ccntctntgg gttagaaata    120 

ccgctntatn gngtatgagg ggatngggcn tacgnnataa tttnctatng ganggtattn    180 

ccgcactant gacnagttct ttctnnggtc catttnnaac nacantnttg acattgntga    240 

tctgcaannc tgtaaaatag tcttncagtg ggcaatnnnt gcacaactgg gttnggtntc    300 

anaca                                                                305 

 
           
             14  
             308  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(308)  
               n = A,T,C or G  
             
           
            14 

agcagacaac ntaatccaag ccatttacca aataantata tgcgatgcac attgaatcct     60 

ggcgctctag atatantgcc ccaaaggaaa gagnacaaag tnttccnccc ntagttctac    120 

natgnctatc cnctatcacc tnctgnttcn naagntttnt aaaaataaat tctcttgtat    180 

ancatccnat atcncaccgg tccaaagcgc aacaatctgc aattcanaan ttccaacaat    240 

cnatntatgn actttcntag gtccggtgtt ctaanatnta atattctaac acttactctc    300 

agatctta                                                             308 

 
           
             15  
             304  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(304)  
               n = A,T,C or G  
             
           
            15 

ngtnaaggga tatttattcc tgttttaaaa ggatacaacc aaggtaggga aggcttcgtt     60 

attggtgatt attcagaaga cctattttct ttacatatgc tatggaaaca atactgtttt    120 

ccgctacaga atacagttta tgattatact tttgtaaatt gcctgctttt cccctgtcat    180 

ctgctaattc caatttgata ctgttctgtg ttcaaaaata cagcatgagc aagctgtaat    240 

ggtgcctgtc gagagtccca gctgcttggg gggctaaggt gggaggatca tttgagccca    300 

ggag                                                                 304 

 
           
             16  
             703  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(703)  
               n = A,T,C or G  
             
           
            16 

ccggtngnct aaaaaggacc agcctaatgt agaaggtggg tatttggacc agaggcttta     60 

gattattatt ttagatccta catatacttt tatcagtaga atgatttcat tnagatgtat    120 

aatgaaaaag ggtaatgcaa aaattatgta atagatacca aattagggaa gtttggcaat    180 

ttcaatggca tatttttagt caaggnacac agatggcagt gccataagca agtctataaa    240 

tatcggctgc agccatcccc ctcattttaa atgttgccct aataatcaat gcagttaaca    300 

agtatattgg ctgtgtgtca tgaaatagtt catgttcaga tggaaatgtt aggttactgt    360 

atggtttatg gagattaatg aaaatgaatg cccaaaaaaa aaannccntt tngnggnggg    420 

tttnnnangn acngggctgg attcaaanca ttggggatnc angnttnaat gngnccccat    480 

ttgnctnaac ttaccttnna nnntgggcnn tnnatngaan angggatnnt gggannaacc    540 

tttnnangnt nnaantgtnn ncttactggn gnaaannncc ntaanntttn nnnntnnnnn    600 

ngnaangggg naannnnnnn ntnancttnt gggggagncn nttntggggn anggggggnt    660 

nnttnnnncn tnnnggccnn nnnnggggcn nnaaantttt tgn                      703 

 
           
             17  
             171  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(171)  
               n = A,T,C or G  
             
           
            17 

tccgcntcta agtaattcat caataacgca tgtccactta atgtgaaaat tggtaccatc     60 

taatanaatc ttcaacatgg cnatccacnc tattccaata atgaaatgca aatttccctg    120 

ccttctttac tanggtcatt tntagattct tgaggaatga gttctactct t             171 

 
           
             18  
             689  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(689)  
               n = A,T,C or G  
             
           
            18 

antnngcttn ggtactaagc agaatcactt ncttgggaac tccatgtaac tngtggcttt     60 

tgtgattgaa atagcatcag taaangtctg accctgtggt aaagacacat atgngcgtgg    120 

accnggctat gtctgacttt gtgctgctca ggacactctc tgtnaccaaa agngagagan    180 

cctggannac ctcanggggt canatgtttg aaggagctgc tgagtatcct ggcaggcanc    240 

anagccttac catcagtttg ctgcatggaa ggctgtgtgc ctctatttcc ctgctatttg    300 

ttgaactccc ttgagctccg gtccttccta agtgagagag atgatcccaa tagcnccaac    360 

ctgagagggc tggggagatg ttngaaggaa agcttggctg gggagctgaa tctggcctgt    420 

ggtacatgct tggtaactgg tggccaggan acccgggngt gtgtnctggg actgtcncac    480 

tctgctgacc agggtattga aagtccccnc tcaaanacac agaatntntc tgaccaaggg    540 

tangtatgan atgacntgtg gagcactttg nataaactgg ttctcatngg nggtcccctt    600 

gaanaggtgc tnnatctgtt caaaaatacg tggctgagct ntanacccng natcctctgt    660 

cagagacatg ggcaggggga ctcaatgct                                      689 

 
           
             19  
             721  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(721)  
               n = A,T,C or G  
             
           
            19 

tatanatact nngctatgct ttctaccctg tgtgcctgga gacctactat ggaaaaanga     60 

tcagccacct taccttctac tgggtacctg ctgtgagtct gcctatgcca caacgattaa    120 

tgangggagg gtacccaagn gacaaanccn acatgccgct tacagccccc gttggatngn    180 

tgctcattca acagtcttgc attcagtagg tgtttgacat cacctactat gtgncaggct    240 

ctatgctang nactggggat acaggagaga ntnaagcgta aagtctttgg tctcaaggaa    300 

tttgcattct agaaagtcta agatgtaata aatgtactgt gggacatgtt aaataagtgc    360 

tataacgaaa tataaagggt ttgggagcaa aaaanaaacc cnnttgtggg gntctntncc    420 

nctctgatga agcttactta cttttaacct tnccttctcc tttaaaggtg tttcctggtt    480 

cccctttcct ttacagattg gttattggtc ttgctgagga gtaggactac aattnccagc    540 

attctnctgg aagccaaagc tgtgctacaa ttgnnccaaa gaagatngta atcttaagcg    600 

cccntaatgg taaaatngta ttaaaangtg gacctttgac aaataaattg nttcgatttc    660 

ngaattccgg gttngnagct tngngntncc aaaaaccctt nggggntccc ttttgggcac    720 

c                                                                    721 

 
           
             20  
             248  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(248)  
               n = A,T,C or G  
             
           
            20 

cttaaacacc ccncccatct ncnccccaga atgagntaan catactcntc nntactgnat     60 

ctccgtatcc gtccctacnc nggnttgtga ggtgtcatta gcngatatta ctcctcatcn    120 

ncatcntgan cannatcccc catcnnccat atgntgatna nnacaaacca tnctattncg    180 

ccgnngaagc cnntcnnttc attggattcn tagaccgcan angtcctnat tcngacacng    240 

aatcggta                                                             248 

 
           
             21  
             298  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(298)  
               n = A,T,C or G  
             
           
            21 

ggtctaaggg atgtgatgng agcatagaat ttanctntat ggncatanta gggacatntg     60 

ctgatntacn tggnctgcgg tcnntgaaag gtggngnatg atgactgatg tcatnagtag    120 

tacnanggac tncgnnanct gggatcnggg nttacnttgt tcatngtnag agtgnnancn    180 

aagtanatgn taggnataaa gatgttncgg gagatgggtc tacaaantct tttnaagatg    240 

ntcatcttga anannatcaa gtgtgnttgg tataatgact atcattatac aatgtcaa      298 

 
           
             22  
             591  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(591)  
               n = A,T,C or G  
             
           
            22 

tcgctagant actattcggc cgcaacgggg agcctgatga ggacgcttat gatatgagga     60 

aagcactttc cagggatact gagaagaaat ccatcatacc attacctcat cctgtgaggc    120 

ctgaagacat tgaataaccc tgggcagtgg ttcttaggca gatactctag atgctttatg    180 

gacaatatta ttttcattgg atgattctgg agctctatta ggagaaaagt aatcatttta    240 

ggtcttaaag acttcaagaa aatacaggtt atcaatttat tttaaatctc attgtttcca    300 

gttagcaata tcatacctat taaagctgtt cattgtaaca aaattcaatc aaaaaggcag    360 

ctaggtcaga aggaaacata ccactctcat ggttcatagt attcactgta tgtatgctag    420 

ggaaaagact tgctccagtc tcctcctcag ttctgtgcct gagaaccact gctgcatata    480 

tttgttttta aattttgtat tgaactgtta attgaagctt taaaagcata tatgaaatgt    540 

ataaatctaa gatgtataat acattattga ctccaaaaaa aaaaacccct t             591 

 
           
             23  
             755  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(755)  
               n = A,T,C or G  
             
           
            23 

gnnnnnngtt nnnnagcngg ttnggtncng actcccnttt atnatgaggg acactgaggc     60 

ttcaagagat taggagactt gttcaaagac acacagctgg taagtgatgg aggcaggatt    120 

taaacctggg tttcactgca tttcccatca ctggctttta gccatgatgc tctactgtgt    180 

aaccctctta attcttgacc tgtggctata aagtatgtat tgagagacag gccctccctg    240 

agataacttt ccagccttga caaaggcaca cccttggttc attccttgga gtgtaggacc    300 

tagattgtga caagcccaga tgagtgtgtc tggcagaggg gagcagatct gaggccacca    360 

tatgtgttca cctagcccta aggagtgcca gcttcgctgg tatttgtaca gcttccatca    420 

ggactgctca ttggccacgt tctttcctct ccctgccacg ttgattaata ctcacataaa    480 

ttaatgctca cattagtgtt caagtatgca aatgagtgct taaaatcatc actcacacaa    540 

tgaccagact gaggatataa cacacaagag cccctctcct ggtaacccca caatcatgca    600 

gatgtgttga cttctctgca ttaccagtct ggtaggcagg gggatatgac agttagaaac    660 

agtctttcan acagcagttc tcaacaccag gtcccttgct gcacaatcga atcacctggg    720 

ggtttaaaaa aatatcatgc cagtcagcca cnntt                               755 

 
           
             24  
             513  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(513)  
               n = A,T,C or G  
             
           
            24 

ctttctaccc aacaagcata gaatatacat tgtatacatc agaaacacgg gacattctcc     60 

aaaatagacc atatgatagg gcacaaaaca agtctcagta aatttaagaa aatcagaatt    120 

atatcaagta ctctctcaga ccacagtgga ataaaattgg aaattaattc cgaaaggaac    180 

actcaaaagc atgcaaatac atggtaatta aataacctac tcctgaatga ttgttgggtc    240 

nacaatgata tcaagaggga aatttaaaaa ttctttgaac tgaacgataa tagtgacaca    300 

gcctatcaaa aactctggga tacagcaaaa gtggaggtaa gaagaaaatt catagcatta    360 

aatgcctata tcaaaaatct gaaagagcac aaataaacaa tctaaggtca ccctcncaga    420 

attggagaaa ctagaacagt ccaaatccaa acccngcaga agaaaagaaa taaccaaatc    480 

cgaacaaaac taaatgaatt gaaaaaaatc ccc                                 513 

 
           
             25  
             574  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(574)  
               n = A,T,C or G  
             
           
            25 

cgatccaaga gattagaanc ccntggagtg gagcatgctt cnctanaatn ccacctgatn     60 

cttggctnaa nacantnngc tctantttgc tttgtgcccg tccacacaan ctaaaaacaa    120 

gggatggggg gaccncnagt gtctaatatn cntaatatcc ntccncnggc aaatgaatac    180 

tttttacaca cttgtanntt ntggagggan ggggtnatna tgaggggaan gggaaaggat    240 

gaggagaaat ccaggatnan angtctcttc gtcctctcna gactncctca cactctntgt    300 

ggtnaccngg gttcgttntg tccaatggca gacattatac tccatantct acccnggctt    360 

nntcgggttg ggacgccann actcccccna gtngtnnccc ccnancagcn atacacaagt    420 

ntgaacgggt tttgtggcca ntcatcgcaa tgaccttntc ctcnactcna agaaaantaa    480 

accccttccc ccngattggt ttctaaatct ttcaccccat ctaaaataga aagcnctnag    540 

tgggangggt tnatcccccc nttaccntta aaac                                574 

 
           
             26  
             185  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(185)  
               n = A,T,C or G  
             
           
            26 

gnacnattgg caatgacnga aagaatttga angatgnaca agtnaaagnn acagtggcaa     60 

agaatcttcn gggcgcgtca aaacaattgg gtgnattaag gacaanctcg gtcancagta    120 

taanctctct ttcncgngga ttantngnca taatcatnat tctgacnngt aggacattnc    180 

caacc                                                                185 

 
           
             27  
             270  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(270)  
               n = A,T,C or G  
             
           
            27 

ttctggggct ctatacaggc tcctatttng atccangcgt gctgatgagt gcacagcacg     60 

atcacatctg gaaaccacca ntaccaccac cactacgcac ntcaccaaaa ctgtganagg    120 

gggcatttca gagacaanaa ttgaaaancg aatagtcntc acgggggnat gcanacattg    180 

accatgacca ggcgctggct caggcagnta aagaggccan agatcaacac cctgacatgt    240 

cngtgaccag agtggtggtc cttacanaga                                     270 

 
           
             28  
             758  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(758)  
               n = A,T,C or G  
             
           
            28 

tgctaggtan aaagttacct ctaagggaag ctctgcagaa gaaatcagtg aaatactctg     60 

aaagccgcaa ttacaatcaa gaggaaccta cttccctcct ggcaaagaaa cccaaggaag    120 

gcgagcggaa gatttacttg gcaattgaaa gtgccaatga actggctgtg cagaaagcaa    180 

aggcagaaat caccaggctc ataaaagaag agctgatccg gctgcaaaat tcataccaac    240 

caacaaataa aggaagatac aaagtcttat agacatccgg aaaaaagatt tttacctgtg    300 

ctggtctatg atgtatgtgg cagttgctgt ctgcagttta caatgtattg tnaatgaaga    360 

ttttttaaat tctatcttgc tgattttttt taaatataan aaactggtac ttggtaaaga    420 

aatctgtccg taattncccc ccaatcagtc caactatatt taaagccacc tgttttcnaa    480 

ttttgatntc ctttaatgtt nactccaata tccatatttt aaatgtcccg gataatatcc    540 

caaaggttta aaaaatggaa atntttgaac ttcnnttgaa nanaataaat tcccatcctt    600 

tangggntnt ccccttnccc gttcttccaa gaaatgtgac cttccccaaa aaagntnatc    660 

cctanctttt tgnttccccc ctgantttct gancccggac antnacgggt ttaaaanttt    720 

ttaaattttc caanncaaaa aaccntntnn ttttttna                            758 

 
           
             29  
             577  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(577)  
               n = A,T,C or G  
             
           
            29 

ctgctaggta ntaanattat ggatccacat tgtnctgagg anacgaanat acttgctgct     60 

gatngaggtg aaaacgatat tgatccntct ggggttttac ggtgtgcact gggtgctgca    120 

cnnacttgtc aaggtttgnt acgtcctctg ggcatctgca aaaggccctg ctctctggag    180 

tgttgtatgt agtgtaccaa aanagtattt atacatccca ccaatcaaaa cacagctttn    240 

ttacctcatg cgaactcatn caaaccaata gaatntcaac atgttctgta ccttanagtg    300 

ctcacttact acctctgaac natactcacg ctgtnntttg tctcttnctt atctttttgc    360 

ntcttgtaat taactctttg tttcccttca tcaaatgtaa tgtanatcgt gatctattaa    420 

aanaaaaatc anggttgcac ttgctacttt naanaaaccg antgtggaaa cattgggtct    480 

naattcacac aggatcngta naactgttgt ggatactgag aaacntttga atgttcctcc    540 

ccttattacc atcccgcaaa aaaacccctn tnntttt                             577 

 
           
             30  
             449  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(449)  
               n = A,T,C or G  
             
           
            30 

tttacccaat aanntatagg cgatagaatt gatacctggc gcaatagata tagtaccgca     60 

aggganagat gaaaaattat aacnaagcat aatatagcaa ggactaaccc ctatnccttn    120 

tgcataatga attaactaga aataactttg caaggagagc caaagctaan accnccgaaa    180 

ccagacgagc tacctangaa cagctaaaag agcacacccg tctatgtagc anaatagtgg    240 

gaagatttat aggtagaggc gacaaaccta ccgagcctgg tgatagctgg ttgtccaaga    300 

tagaatctta gttcaacttt aaatttgccc acanaaccct ataaatcccc ttgtaaattt    360 

aactgttagt ccaaagagga acagctcttt ggacactagg aaaaaacctt gtagagagag    420 

tcataaaaaa aanccctntn gggnnnngn                                      449 

 
           
             31  
             500  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(500)  
               n = A,T,C or G  
             
           
            31 

tcntggaccc nggtccccnn gngancaaan aagaagggcn ngnttncatn gaaaancctg     60 

tgattntcgc cccggtncag gtgttnannt atggcccncn cncatctggt atacgccnaa    120 

acaatntant tttacaatnn gtnccccanc aaacaangtt cgtngnnttn actaggtagt    180 

taatcccncc ccatgttcaa ataaagggcc cgcgntncna ataaggaanc cnccccgant    240 

ggggtccccg aggccctctc cttcataaaa nncattcaac ttccctcccn ctannaaagn    300 

aattnttcna atttttnaaa cactccctgt ccanggggac tttnccccca ntanctgaaa    360 

aaatngcntg acgttcccct tcggcctaag ggcncaactt anttnncccc caanacccgn    420 

gggnnaggnn naaactcccc tngaagggaa cnactcgcnt aaaaanggaa taatcncccc    480 

cnaattattc cctncccggg                                                500 

 
           
             32  
             426  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(426)  
               n = A,T,C or G  
             
           
            32 

gtctatgatc acatctgacg ctattcctat ccccttcctc cccgggacct tttccccttc     60 

ctccctggga ccttttcccc ttcctgttta anaanccagg gctgcctgga ggaagctttg    120 

tcagatctag tggaatgtga cctccctgga atatgtgccc aggggtttgt ctaagcagtt    180 

tcaggctatg gcctttactc catctggtcc ccatccctct tatctctctc atgtgtggct    240 

gcacctggac gcttggacca tagctgtcac agccccctgg ggaggaaccc actccttggc    300 

catntcagcc tgtgcaatgc aaggctcttg tttgatctgt gtgctgacan aaagcccagc    360 

ttccttaaga acttttcatg tggaacactt tggttttgan aagaaaataa atcanaaacc    420 

attaaa                                                               426 

 
           
             33  
             375  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(375)  
               n = A,T,C or G  
             
           
            33 

ngttgcacct attggccngc tggtctcgac tcctgacctc gttatctgcc tgcctcggcc     60 

tcctaaagtg ctgggattac aggagtgagc cacagtgcct ggcctgtcaa gacttctctt    120 

aagttaactt cctgagaagt gatgtctaaa agtatctttg ctggtgtgag aactccagtt    180 

tccaacacat attatttccc tcaactattt ggaatatttt agaattttaa ttccaaagga    240 

ttagtttgaa tacaagtatg ccacataact cagttttcgc catcttncat ttcttaacag    300 

tgtaaattaa aagctaataa tcataataat aaagtgcatt taattatctt cgaaaaaaaa    360 

aaancccttt tgggg                                                     375 

 
           
             34  
             809  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(809)  
               n = A,T,C or G  
             
           
            34 

ttgcacatgc tggccaggat ggtctcgatc tcctgacctc gtgatctgcc cgcctcggcc     60 

tcccaaagtg ctggaactac aggtgtgagc caccacgcct ggcagctttg tgttcttttc    120 

tttctgtgat cttgccttag atcacacaga taaaacatga caggacctgg accttaacac    180 

agtttggctc tcaatcctgt tctcataacc acnactgcct tcatttatct gtgtcatcct    240 

cagacctgac acatagtagg tgctcagtca gtgttcacta agtaaatgat gaccaagaac    300 

tctttgactg ggtccaaggt gcttatccca atacttcgcc atggctacct ccctcattcc    360 

tcagctgact tgctctctct agcctggctg ctcctatttt atttcctaaa catggaccca    420 

tggcaataag tttaaancta acangttgat acggtaccca tccataattt aatnaattnt    480 

ggggctcatg caaccncaaa aaccagaacc caaaactacc tgtncncaaa caacaatcat    540 

tttnggtngg gatcccntnc tngcttggnc ccttttttta aaatgtccat tccccccgga    600 

ctttaagaaa ttgaaggaat ncccggaaan tattgttanc gggccccctt nagngaaaaa    660 

ggtggcnctc cnnncggggg ccctccctgt ccctgaaatt tnaaaacccc cctcccnntt    720 

taaanccctt aatcccggnt aacancnaaa naaaattcta gggcccaaac ccannggttt    780 

ggttttaaaa aaccntntat ttttttnat                                      809 

 
           
             35  
             192  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(192)  
               n = A,T,C or G  
             
           
            35 

caccttattg ggatacagca gtgaattaag ctattaaaat aagataatga ttgcttttat     60 

accttcagta gagaaaagtc tttgcatata aagtaatgtt taaaaaacat gtattgaaca    120 

cgacattgta tgaagcacaa taaagattct gaagccaaaa aaaaaaaccc caanggggnt    180 

nnttttnaaa aa                                                        192 

 
           
             36  
             368  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(368)  
               n = A,T,C or G  
             
           
            36 

ctgctagtac caantattat ttaagantac ttttcactac tcctaaataa tgacacagat     60 

acgtttgtct tacacatttc actttattgt caagttatta gtatgtttat tttcaaaagt    120 

tattttttgc aatttctttt tattattccg tactttttaa atttacttca ttatcacgtc    180 

ttcctttatt ctttttaaat agtttttgct tttgttattt tgttttccct tttttactct    240 

tggtttgtaa tacctctttc cttatttgct cctttctcat ttgatctcaa tgttaatcca    300 

actgttttcc acatctgatt cactaaaatt ttagcccaaa aaaaaaancc cntttngggg    360 

gngntttt                                                             368 

 
           
             37  
             219  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(219)  
               n = A,T,C or G  
             
           
            37 

ggccccattt cactctccat antggcnctt nctngaacag gcgtnctgga tnagtgcaca     60 

tacnatccca tcnacntgca cctatancnc ttccactacg cacatcacca aanctgtgaa    120 

agggggcntn tcnttagaca cacaattgca gaatngacnn cncancccgg gggannctcn    180 

angttcaccn tgnagcaggn gctggctcan gctnttata                           219 

 
           
             38  
             198  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(198)  
               n = A,T,C or G  
             
           
            38 

tcgatacagg gncagatctg ggagccaggg cgttgctgat gagttgcaca gacgatcaca     60 

tctgaaacca ccagtaccac caccactacg cacatcacca aagcgctggc tcnggcaatt    120 

aangaggcca aagagcanca ccctgacatg tcngtgaccn ttgtantggt ccntaangac    180 

acngacatcg cctccaca                                                  198 

 
           
             39  
             560  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(560)  
               n = A,T,C or G  
             
           
            39 

tttnnatcng nacagctagt cctntaaant aatgacttca tagaaatggc attataattt     60 

ttaagttgat actctacagg tagctattga tataattagt tttaataaaa catgctgcaa    120 

ccatggtata caacaaaaat acatttcttt ggtgattgaa attaaggccg tatttacaat    180 

gacttaatat aagactgact tttatcctgc ttcataactt gtatggagaa ctcaccaaga    240 

aagaattcaa tactgtgaaa tatgcagcaa gaagattggt ctttacctag gctgtgtttc    300 

ctaagctctg agttttcagc accagtagat ttgtattaaa agaaaaaaaa atggggcctt    360 

agcttctggc ttttaatttt gccagctaag gacataaaac aaaantaanc aancaaaanc    420 

aaatagccat ntgctatcag catcattatg taaaagaaaa tntattttag cccctaaaat    480 

taggaagaat gtaatctcag aataaaggtt gtcatttaag ttgaataaat atntagcttt    540 

cgaaaaaaaa aanccccttt                                                560 

 
           
             40  
             421  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(421)  
               n = A,T,C or G  
             
           
            40 

atacagggca gcgtgttagg tgaccacacc aggagcctca gcctcggtcc ttctcagccg     60 

tcgggataag atccaggcat gncttttaaa tctcagaggt agcagtaaac ttttcantnt    120 

tgcngttagc aagtgtgtgt ttgccaataa anccccatta tactaatgtg cctanttaat    180 

gttcagggaa natctgcttc cactgtgtnc cnaggggtgn catgaactnt gtgagnagcc    240 

ccncnnctgg agggatgaat gctgngttaa ctacngctat cacggatngt gtgntgtgaa    300 

naatacatcn acatnaatnt tanntgctct gnaanttccc ttnttatntg tcaagtaact    360 

ntttgtaaaa ntnntnctcc caanttatta cngtgattac taatnnattn gtnccatgtt    420 

t                                                                    421 

 
           
             41  
             411  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(411)  
               n = A,T,C or G  
             
           
            41 

aggtagaggt tgtgcatgtt gtccttttta tctgatctgt gattaaagca gtaatatttt     60 

aagatggact gggaaaaaca tcaactcctg aagttagaaa taagaatggt ttgtaaaatc    120 

cacagctata tcctgatgct ggatggtatt aatcttgtgt agtcttcaac tggttagtgt    180 

gaaatagttc tgccacctct gacgcaccac tgccaatgct gtacgtactg catttgcccc    240 

ttgagccagg tggatgttta ccgtgtgtta tataacttcc tggctccttc actgaacatg    300 

cctantccaa cattttttcc cagtggagtc ncatcctggg atccagtgta taaatcccaa    360 

ttatcatgtc ttgtgcataa attcttccca aaagggatct ntaatttttt g             411 

 
           
             42  
             408  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(408)  
               n = A,T,C or G  
             
           
            42 

ggctcccctc cctaactctc taagtacttc ccttacccac tcagtgtggt gatggcacct     60 

ccctgaatct cctgacaaat gcgaacagga actcctattc atcaggagcc aacttgataa    120 

ctganaagat tcctctctca tttatcagcc tttgattatc tttttgtgtc tcttactatt    180 

tgcgcttagc gagaaaaata aagaggtttg aacaattaag aagtaacaaa gagctcatag    240 

ttcacaaaga gcaantcaaa ggatgtctgg aatatttgaa catacaactg cctttggcat    300 

gaggtggcct acatacattc tcaggggcag gataggctgg nanagctgat caagctgccg    360 

ggaaagctga agcaaaggca gggttggntg gaaatcaaaa tntctctt                 408 

 
           
             43  
             275  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(275)  
               n = A,T,C or G  
             
           
            43 

tccctaactc tctaagtact tcccttaccc actcagtgtg gtgatggcac ctccctgaat     60 

ctcctgacaa atgcgaacag gaactcctat tcatcagagc caacttgata actgagaaga    120 

ttcctctctc atttatcagc ctttgattat ctttttgtgt ctcttactat ttgcgcttag    180 

caagaaaaat aaagaggttt gaacaantaa gaagtancnn ggagctcnta gttcanaagn    240 

agcaagtcaa aggatgtctg gangatttga agggt                               275 

 
           
             44  
             246  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(246)  
               n = A,T,C or G  
             
           
            44 

tttggtccca agcacatttc acaaangaga atttacacct agcacagctg gtgccangan     60 

atntcctang gacatggcca cctgggtcca ctccagcgac agacccctga caagagcagg    120 

tctctggagg ctnantngca tggggcctan tntcntcaat cnaatgagcc ccnantgcta    180 

ctgcgccccg ggggctccca cggcctgggc nnctttcntg caactgnaaa aggatagngg    240 

tatttc                                                               246 

 
           
             45  
             345  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(345)  
               n = A,T,C or G  
             
           
            45 

tttggctccg tgggacgttg tantgtgcnc agacatttcc aagggaaatt ctaaacagtc     60 

accctnccct tttgcattcc cccaaatctt aagtgtatac ataaaaccct gggtacatat    120 

tgtngtggta atagaaggga attggnnaaa cngtacactt gttatatgga antnactgtg    180 

gccacctaca aaagacaagt taacaaactg tcntggaggc tgtngntgcc canccagggc    240 

cgctgcnttt tgacaacatt cccaccctgg ccactcagca canttcatgg caggtcatgt    300 

ctntncactg anacntttnt ganacttttt catatagcan aatcc                    345 

 
           
             46  
             969  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(969)  
               n = A,T,C or G  
             
           
            46 

aattgcagtt ctttcttgcc tttaacaaca ttagggcctt tagaatgagt acctggtgct     60 

gtccttccaa ctctgtgatt ctctgattcc atcctcattt ttcaccatca ctggtgtact    120 

ggcaagaacc antatgagat ttgaggaaaa atacttggat tactcttttt taaaaaaaat    180 

tatttagata taattcccat accatacaat taaccttttt atgtgtataa ttcagtattt    240 

ntagtatatc cacaaagttg tgctaccatc accactatcc gattccagag cttgtcatca    300 

tacaaaaaaa aaaaccccan agtnanttcc tttcaaaacn ctttnngttn ttcnttntnc    360 

ccntgtngcn tctagnncng ngggntnnct tttgtcnntn tcnccctncn ctcatcntnn    420 

cnggtctctg ctcngngnnn cgntntgnct tnnantcgct gctnntcntg tattccccgc    480 

nctngtnnng tctgcnncgt agccagtggn cctcctgntn ccnncngntt ctntntncgg    540 

cacanntcca nccanctgcc atnagtnana nnatctctnt tcnncanctg ntnncagnnt    600 

tgtcntcntc tccgtnccnc cngcngctnn ctcnttncgc nctggnngnc antcgtacct    660 

ggcttttatc cccctntccn nctnttctng atggnntctc ntctcnacac ctgncgttac    720 

gnntctcntn tnncnnnann cgttnctntn tnncttnccg ncngccatct nagctcannc    780 

tggngcgant cncgctctgn gtatcagtca tntanagann ngngnntgtt nccnncgcgn    840 

nntgagannc ccncccnctt cgcatnacgt angtgncttt ntnnatctgc tcgtcgtctc    900 

nctcatatcc nccatgctgn catganactc cntantctnn cgcnnttctn ncgttccctc    960 

tgcccttnn                                                            969 

 
           
             47  
             361  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(361)  
               n = A,T,C or G  
             
           
            47 

ggccactaag caggtcttac cnaatttaag aanattgaan tcctatcaag tatctcttct     60 

gaccacaatg gtatgaaact agaaatcagt aacaggagga aaattggaag attcacaaat    120 

ntgtggaant taatcaacnc atgagcaact antgagtcna agancanatc aaaagggann    180 

tcaaaaactc tcttgaggtg gatgagaatg ganatacaac ataccngaac tcatgggatg    240 

tatcacaagc ngtgctaagg gggaagttta agtnctagat gtctanatta ngaaagggaa    300 

agatctcana tanacnaccc agcnttncnc ctcgaanaac tagaaaaact aagaaaaaac    360 

t                                                                    361 

 
           
             48  
             364  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(364)  
               n = A,T,C or G  
             
           
            48 

atgatgacca catntagatg gcacatngat gaggacttta atctttcctt aaanacaata     60 

atgtgttctt ttttctttta ntcacatgat ttctaagtan attttncatg caggacactt    120 

tttcaacctt gatgtacant gactgtgtaa aatttntctt tcagtggcaa cctctataat    180 

ctttannata tggtgagcat ctngtctgtt tagaanggga tatgacaata aatctatcag    240 

atggaaaatc ctgttacaaa gtataaaagc tttagtaatt tactcagtgt ggtggtttta    300 

tcctttttgc tttttctccc ttggtctata atgaaattgt tacagcagtg caaaataaaa    360 

tcct                                                                 364 

 
           
             49  
             703  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(703)  
               n = A,T,C or G  
             
           
            49 

atggggaatc aaacaatgtt aaaaggctan taatacttat aggttttatg attcaattta     60 

ctatgtgttt aaaattgttt tttgaaaaaa ttgagttatg tcnctaaaac tgagtctnta    120 

cagctcaaaa atgaagaaat acntatctcc gataagcata ttatgtgaat ttcaacatcn    180 

ctattgagaa aaggaatata aatttgaatg aaaatgaaac tctatctttc tatatcacat    240 

tgcataggtg taggctagtg agtactttga tgtaaattgc tgtatctttt gaggcntcna    300 

tttggcnata tagatcagaa ttttaaatcn gcatactttg tttgccagaa atctatcagg    360 

accacttgta ntnattttgt tnaaaggaat atcnaacnct tggatgttca ncncagtatt    420 

gattgtttta naagaaggaa anggagaaag ggaggagaat ggaaganana aanggaggga    480 

ggaanattgg aaccnttgac atntgtgata gcatnggatt tgctnaacac nctatantat    540 

acccctngca tggganaagc atgcacnctn aaacaaggac nngttngatg gntctacnnt    600 

ttgacntcag atnnaantaa atnaaaaaaa aaancccccn cctctttgnn ttcctntcnn    660 

cgnnnnannc ntctccccnc nncgnccnnc ncccgccacc ntn                      703 

 
           
             50  
             413  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(413)  
               n = A,T,C or G  
             
           
            50 

tcttggctgg ttgagtattc aanaatcagg cacggagaag tggggtggat gcaaaccaac     60 

tgaccactgt ggcaccacca gcagtttcag ttttcatctt gantgtcnag aggaaatatc    120 

taatcttaca actcnttagg ggcctggctc agtggctcat accttgtntt cccancactt    180 

tgggangccg angcnggcnt atcacccgca ngtcaggatt ttgagaccac cctggccaac    240 

ntggtgaaac cccatctcta ctantcaata caaancttag ctangcgtga tggcatgcac    300 

ctctaatccc acttacttgg gangctgagg cagcganaat cacttgtaac ccggaaggca    360 

nacgttgcat ntgagccaag atcgtgccac tgcactccat cctgggcttt cta           413 

 
           
             51  
             252  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(252)  
               n = A,T,C or G  
             
           
            51 

gttacagaca aggnttntag aatatcttat gttttatgct ctgtaagttc aaagaagnta     60 

gcagaaaaca taagcatact gaaaagagaa acagaagcta ttttttaaat acctatgtga    120 

aatctctcta tntgaaacaa aaaatacact ggatggatta gacactgcag aaggaaaatt    180 

tggtgaactt gagatcttat aaataaaaat tatccaaaat gaagtgtaga gtgaaaaaaa    240 

aaaancccct at                                                        252 

 
           
             52  
             875  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(875)  
               n = A,T,C or G  
             
           
            52 

agaaacgaga atgganattc aaatacgtcn gccgggcttg gtggattaga cctgtaaccc     60 

naacactttg ggaggnctag gtgggcggat caccngaggt cnngagtacg ggaacancct    120 

ggcaaaaacc ccntctttan tctgngaaaa cncaactcta ctaaaanaac tactcttaga    180 

tnggcgtngn tgcgcctgcc tgttntccca gatacnnttt naggctgang tggggataan    240 

tnctttaaca tgggaagtgg aagttgcact gatccaatgt ctccacactg cantccagcc    300 

tgggttangg aatgagaccc cncncacgga aaggacaata aaaanccccn nnggnnttnn    360 

tttttaangg cctcttgntc nttttcttnt antgcncgcc tncgcnnncn ttgntntgtc    420 

gantcnnntg cnnttntttc ttcnncctcn ancctgcttc tnntcnnttc gccnntnnac    480 

ngcttccccc ntnctctagc acttnnnttc tntcgntccn nnatctccnn cttntctnnn    540 

ccgctcgcgt nnnccntnan ctcgnntcnt nccctttctt cncngcnncn ntttcgncna    600 

gatcgtncgn ctctatctac ttctntccnn gntntanata tngatnttac attntgctcn    660 

atnacccatn annncntcta tgtttatann ngtnnnnccn ttcaacnnnn cnttatgagn    720 

tcttnactca gctctncgtt gntnttccna ctanngttgn ncntncatgt nctgtcncgt    780 

ancnctctnc tcntcncngt cntgagacna atctctatnt atngnttatn cctgcntnct    840 

ganctncacc gngatctcgg cnntntcttc tcaag                               875 

 
           
             53  
             182  
             DNA  
             Homo sapien  
           
            53 

ccagaagaag ggctacatat ggactcatgt tgggcctact cctgcaataa caattaagga     60 

atcagttgcc aaccatttgt agttcacaaa ttaaaactgg gtttccaggc ctggtgtggt    120 

ggctcacgcc tgtagcccca gctattgcac cactgctctc caagctgggc aatggagtca    180 

ga                                                                   182 

 
           
             54  
             329  
             DNA  
             Homo sapien  
           
            54 

catgatgcga gactggacat ctctcctacc ccatgtacac ttcagctgag caggcagaat     60 

tagagagtca ggactagaag ttcagtctag ggatcaaata ataatagtag ctaatgttta    120 

aaggtaccta agatccgcca ggagacatac tcagtatagt tccgtggttt gccacatttc    180 

atcttatcca gtagcacagg tgaaatttgt cttatgtgta tactgaggaa aaacaagtcc    240 

ctctgatacc agcagccaat aaatgacaaa gctgggatag aaacttactt cattctaacc    300 

cgagagtccc tgttcttgca tggggcaca                                      329 

 
           
             55  
             312  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(312)  
               n = A,T,C or G  
             
           
            55 

actcaactcg tttgagctat aggaatnggc cattcgnngt ggctcanacc tgtaatccca     60 

gnatttnggg anacctcact aggatcacnt gaggtcagga gttcaagacc agcctgtcca    120 

acatggngaa accccatctc tantanaaaa tacagaaatt atccaggtgt ggtggctggc    180 

acctgtaatc ccagctactt gggaggccaa ggcatggaaa attgtctgaa cctgggaagt    240 

ggaggttgcg gtnanctgan atcatgccat tgctctccag cctcggccac anatcaagac    300 

cctatctcaa aa                                                        312 

 
           
             56  
             565  
             DNA  
             Homo sapien  
           
            56 

acaatttcac acaggaaaca gctatgacat gattacgaat ttaatacgac tcactatagg     60 

gaatttggcc ctcgaggcca agaattcggc acgaggggat ccaacgtcgc tccagctgct    120 

cttgacgact ccacagatac cccgaagcca tggcaagcaa gggcttgcag gacctgaagc    180 

aacaggtgga ggggaccgcc caggaagccg tgtcagcggc cggagcggca gctcagcaag    240 

tggtggacca ggccacagag gcggggcaga aagccatgga ccagctggcc aagaccaccc    300 

aggaaaccat cgacaagact gctaaccagg cctctgacac cttctctggg attgggaaaa    360 

aattcggcct cctgaaatga cagcagggag acttgggtcg gcctcctgaa atgacagcag    420 

ggagacttgg gtgacccccc ttccaggcgc catttagcac agcctggccc tgatctccgg    480 

gcagccacca cctcctcggt ctgccccctc attaaaattc acgttcccaa aaaaaaaaaa    540 

aaaaaaaaag atgcggccgc aagct                                          565 

 
           
             57  
             798  
             DNA  
             Homo sapien  
           
            57 

ggaacaagta gaagggaaga gggaaatgga gagcatcctt atgactttac aaagggtgga     60 

aatgaggatg gagggataca gaagtctgca cagctgtaaa ggttttatag atgtctttgc    120 

cttcccttct gaggaaggga agaagtaatg aaagcacatg tgaataaccc cttccatccc    180 

attcacagca tcgcactccc agtccttaag gcaaagggag gcagtgctga agcattggtg    240 

gtgcagtgta aagagacaag acctgatcat ctgatcacac ttgtgccaac ttgattcata    300 

ttgggcatta ctaacaaccc ctggtcaagg taaataggtt gaacaatcaa taacattatc    360 

cctgcctgca tacatgtgaa caaaagctat agaggacatg caaattctac agtcattcct    420 

catatgcttt agacagagtg cagctactgg aatcttccag atttcagtgt tttaaaatca    480 

gagctctgaa tacacaaaag gaaagagaaa tggagcagct gacatatttt aagctcacag    540 

tgatactcag tgacaggagc acagagctct aatgtccaca ggatgttgta gggtagggtc    600 

tctcagtaaa tcaagtccct tacctatgtt ctgacactga ggctcttgga gctatgggtt    660 

agaaatccag gaggcaatat gtctttattc taatgaagtc ctcatcttgc actcagaggc    720 

ccactagttt gcccttctat atattaagta aaaccaagag aaattaaaaa aaaaaaagcc    780 

ctatagtgag tcgtatta                                                  798 

 
           
             58  
             729  
             DNA  
             Homo sapien  
           
            58 

aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa     60 

agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac    120 

gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga    180 

accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa    240 

aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc    300 

tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc    360 

attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca    420 

gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca    480 

gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt    540 

gggccctcta gatgcatgct cgagcggccg ccagtgtgat ggatatctgc agaattcggc    600 

ttgtaatacg actcactata gggctttttt ttttttcggt ttgaggggga atgctggaga    660 

ttgtaatggg tatggagaca tatcatataa gtaatgctag tcttatcctg tgtgaaattg    720 

ttatccgct                                                            729 

 
           
             59  
             730  
             DNA  
             Homo sapien  
           
            59 

aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa     60 

agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac    120 

gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga    180 

accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa    240 

aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc    300 

tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc    360 

attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca    420 

gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca    480 

gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt    540 

gggccctcta gatgcatgct cgagcggccg ccagtgtgat ggatatctgc agaattcggc    600 

ttgtaatacg actcactata gggctttttt ttttttcggt ttgaggggga atgctggaga    660 

ttgtaatggg tatggagaca tatcatataa gtaatgctag tcttatcctg tgtgaaattg    720 

ttatccgcta                                                           730 

 
           
             60  
             623  
             DNA  
             Homo sapien  
           
            60 

gactccaaga gaagactagg aagtagccct cgttctccag ggcacccaaa ataccagcct     60 

ttattgtctg catgatttta ggggatatgg ggagggaaca agtagaaggg aagagggaaa    120 

tggagagcat ccttatgact ttacaaaggg tggaaatgag gatggaggga tacagaagtc    180 

tgcacagctg taaaggtttt atagatgtct ttgccttccc ttctgaggaa gggaagaagt    240 

aatgaaagca catgtgaata accccttcca tcccattcac agcatcgcac tcccagtcct    300 

taaggcaaag ggaggcagtg ctgaagcatt ggtggtgcag tgtaaagaga caagacctga    360 

tcatctgatc acacttgtgc caacttgatt catattgggc attactaaca acccctgggc    420 

aaggtaaata ggttgaacaa tcaataacat tatccctgcc tgcatacatg tgaacaaaag    480 

ctatagagga catgcaaatt ctacagtcat tcctcatatg ctttagacag agtgcagcta    540 

ctggaatctt ccagatttca gtgctttaaa atcagagctc tgaatacaca aaaaaaaaaa    600 

gccctatagt gagtcgtatt aca                                            623 

 
           
             61  
             376  
             DNA  
             Homo sapien  
           
            61 

gcatgctcga gcggccgcca gtgtgatgga tatctgcaga attcggctta gcggataaca     60 

atttcacaca ggatccatga ctcagctatt aaggctctgg ccttggatcc ctatgaggaa    120 

tattttacca caggttcagc agaaggtaac ataaaggttt ggagattgac aggccatggc    180 

ctaattcatt catttaaaag tgaacatgct aagcagtcca tatttcgaaa cattggggct    240 

ggagtcatgc agattgacat catccagggc aatcggctct tctcctgtgg tgcagatggc    300 

acgctgaaaa ccagggtttt gcccaatgct tttaacatcc ctaacagaat tcttgacatt    360 

ctataaagat tggggt                                                    376 

 
           
             62  
             539  
             DNA  
             Homo sapien  
           
            62 

atgactcatt gtttctctgc ctttccgtgt gttacaggtg ggctgatccc cctgcagcca     60 

gtttcccata agcaactgac ttccaactgg gaatgtctcg ggggataatg ggggtgggga    120 

tatggaagta tagagaaaac ataagaaaat actgggtgta tacacctttc tctctctgag    180 

tatgatgaca atgtgatagt cagtgtggca tctgcgactc cagcttgtgc ctggcatgta    240 

caccctagct ccagcttccc ctgggagact gtgcatctcc tggctccact aacaccacct    300 

tcttctgacc ttccagccta gagatgatga ctctgccagc ctagatgggc tctgggttgt    360 

ctccctattc ctgtttgctt tgtagatttc ccattatgct gtcaccaact ccccagccta    420 

agccctctct attttaaatt ctcaagtgga ttatgttcct gattagtccc tgactgatat    480 

accactctcc tcatgatctc tgattagttt tcctgttagg ttgttgcagt aaaaaaaaa     539 

 
           
             63  
             304  
             DNA  
             Homo sapien  
           
            63 

ggcttagcgg ataacaattt cacacaggac gactccaagc tgggaaggaa aattcccttt     60 

tccaacctgt atcaattttt acaacttttt tcctgaaagc agtttagtcc atactttgca    120 

ctgacatact ttttccttct gtgctaaggt aaggtatcca ccctcgatgc aatccacctt    180 

gtgttttctt agggtggaat gtgatgttca gcagcaaact tgcaacagac tggccttctg    240 

tttgttactt tcaaaaggcc cacatgatac aattagagaa ttcccaccgc acaaaaaaaa    300 

aaag                                                                 304 

 
           
             64  
             226  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(226)  
               n = A,T,C or G  
             
           
            64 

atgatgatga ccatgtggac agccaggact ccattgactc gaacgactct gatgatgtng     60 

atgacactga tgattctcac cagtctgatg agtctcacca ttctgatgaa tctgatgaac    120 

tggtcactga ttttcccncg gacctgccng caaccgaagt nttcactcca gttgtccccc    180 

cagtagacac ntntgatggc cgaggtgatg gtgtggttta tggact                   226 

 
           
             65  
             225  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(225)  
               n = A,T,C or G  
             
           
            65 

taccaacaga gcttctgaaa cagataccat agcattggag agaaaaacag ctcacagtct     60 

gaggaagatg atattganag aaggaaagaa ttgaaagcat cttgaagaaa aactcagatt    120 

ggatntggga ttggtcaagt cggccggata atattccccc caaggagttc ctctttaaac    180 

acccgaagcg cacggccacc ctcagcatga ggaacacgag cgtca                    225 

 
           
             66  
             240  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(240)  
               n = A,T,C or G  
             
           
            66 

ccagcatggt ggccgtnatg gatagcgacc cacangcaag ctgggctttg aggaattcaa     60 

gtacttgtgg aacaacatca aaaggtggca ggccatatac aaacagtacg acactgaccg    120 

atcagggacc atgtgcagta gtgaactccc angtgccttt gaggcagcan ggttccacct    180 

gaatgaacan ctctataaca tgatcatccg acnctactca gatgaaagtg ggaacatgga    240 

 
           
             67  
             504  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(504)  
               n = A,T,C or G  
             
           
            67 

cacgaggaga gatngcatct gctatatatt ccacngatac atgtgagtna ctgatagaaa     60 

aaatcgcnnc ggngaacact gncaccggtn ccggcccccg gtactacagg gatctcntca    120 

gacttcaccg tntactacaa ngtaagcncc ctttaagaat gtcacggagt atgatgggca    180 

ggatgcctgc ggctccaaca nctggaacnt ggtggacgtg gacctcccgc ccaacaagga    240 

cntggagccc ggcatcttac tacatgggct gaanccctgg actcagtacg ccgtttacnt    300 

caaggctgtg accctcacca tggtggagaa cgaccatatc cgtggggcca agagtgagat    360 

cttgtncatt cgcnccantg cttcngttcc ttccnttccc ttggacnttc tttcggcatc    420 

aaactcctct tctcagttaa tcgtgaagtg gaaccctccc tctctgccca acggcnacct    480 

gagttactac tttgtgcnct ggca                                           504 

 
           
             68  
             462  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(462)  
               n = A,T,C or G  
             
           
            68 

tggatggcag ggggagaaag gaaaagcaaa acactccagg acctctcccg gatctgtctc     60 

ctcctctagc cagcagtatg gacagctgga cccctgaact tcctctcctc ttacctgggc    120 

agagtgttgt ctctccccaa atttataaaa actaaaatgc atnccattcc tctgaaagca    180 

aaacaaattc ataattgagt gatattaaat anagaggttt tcggaagcag atctgtgaat    240 

atgaaataca tgtgcatatt tcattcccca ggcagacatt ttttagaaat caatacatgc    300 

cccaatattg gaaagacttg ttcttccacg gtgactacag tacatgctga agcgtgccgt    360 

ttcagccctc atttaattca atttgtaagt agcgcagcag cctctgtggg ggaggatagg    420 

ctgaaaaaaa aaaancccct tttttngtnt nttttaaaaa aa                       462 

 
           
             69  
             357  
             DNA  
             Homo sapien  
           
            69 

agaagtcttc ctgagccttc catgtatcct cggtgcccgg ggattaacca gcgttatcaa     60 

ccaaagctaa aggatgatga ggttgctcag ctcaagaaaa gtggagatac cctgtgggac    120 

atccagaagg acctaaaaga cctgtgacta gtgagctcta ggctgtagaa atttaaaaac    180 

tacaatgtat taactcgatc ctttagtttt catccatgta catggatcac agtttgcttt    240 

gatcttcttc aattgtgaat ttgggctcac agaatcaaag cctatgcttg gtttaatgct    300 

tgcaatctga gctcttgaac aaataaaatt aactattgta gtgtgaaaaa aaaaaaa       357 

 
           
             70  
             226  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(226)  
               n = A,T,C or G  
             
           
            70 

atgatgatga ccatgtggac agccaggact ccattgactc gaacgactct gatgatgtng     60 

atgacactga tgattctcac cagtctgatg agtctcacca ttctgatgaa tctgatgaac    120 

tggtcactga ttttcccncg gacctgccng caaccgaagt nttcactcca gttgtccccc    180 

cagtagacac ntntgatggc cgaggtgatg gtgtggttta tggact                   226 

 
           
             71  
             477  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(477)  
               n = A,T,C or G  
             
           
            71 

agcagacaag ccacaattaa catagggtac aattgggtca tgtagctcat gggaaatcca     60 

cagtcgtcaa agctatttct ggagttcata ctgtcaggtt caaaaatgaa ctagaaagaa    120 

atattacaat caagcttgga tatgctaatg ctaagattta taagcttgat gacccaagtt    180 

gccctcggcc agaatgttat agatcttgtg ggagcagtac acctgacgag tttcctacgg    240 

acattccagg gaccaaaggg aacttcagat tagtcagaca tgtttccttt gttgactgtc    300 

ctggccacna tattttgatg gctactatgc tgaacggtgc agcagtgatg gatgcagctc    360 

ttctgttgat agctggtaat gaatcttgcc ctcagcctca gacatcggaa acacctggct    420 

gctatagaag atcatgaaac tggaagccat attttgaatt ctacaaaata aaattga       477 

 
           
             72  
             374  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(374)  
               n = A,T,C or G  
             
           
            72 

ccaagccaga ttgtcactcc agctgatctt ctttgatggt gaagaggctt ttcttcactg     60 

gtctcctcaa gattctctct atgggtctcg acacttaact gcaaagatgg catcgacccc    120 

gcacccacct ggagcgagag gcaccagcca actgcatggc atggatttat tggtcttatt    180 

ggatttgatt ggagctccaa acccaacgtt tcccaatttt tttccanact cagccaggtg    240 

gttcgaanga cttcaagcan ttgaacatga acttcatgaa ttgggtttgc tcaangatca    300 

ctctttggag gggcggtatt tccanaatta cagttatgga ggtgtgattc aggatgaccn    360 

ttttccattt ccaa                                                      374 

 
           
             73  
             597  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(597)  
               n = A,T,C or G  
             
           
            73 

ccaagggatc tgtaaagaat atatacttga gtggtgtgtg ttatcagata aagcaccctg     60 

tatcacagac tggcaacaag aagatggtac cgtgcatcgc acctatttaa gagggaactt    120 

agcagagagc aaatgctatt tgataacagt tactccagta tatgctgatg gaccaggaag    180 

ccctgaatcc ataaaggcat accttaaaca agctccacct tccaaaggac ctactgttcg    240 

gacaaaaaaa gtagggaaaa acgaagctgt cttanagtgg gaccaacttc ctgttgatgt    300 

tcanaatgga tttatcagaa attatactat attttatana accatcattg gaaatgaaac    360 

tgctgtgaat gtggattctt cccacacaga aatntacatt gtcctctttg actagtgaca    420 

cattgtacat ggtacgaatg gcagcataca cagatgaagg tgggaaggat ggtccaaaat    480 

tcacttttac taccccaaan tttgctcaag gganaaattg aagccatant cgtgcctgtt    540 

tgcttancat tcctattgac aactcttctg ggaatgctgt tctgctttaa taagcga       597 

 
           
             74  
             257  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(257)  
               n = A,T,C or G  
             
           
            74 

tggtaaaggg taatagccag agnntagaac cttgangaga tgcggccaan gattctttat     60 

atctgaaccn agatgtnaaa naagaaaatg ctttgaggct ttctaagcga tcctcctgtc    120 

taatttncac ctttgtctgg atgcacactt ctgaccncgc tgccacaacc tgtggggtct    180 

gatgtgtccc ttgatgggtg cggccctcag ggactgcacc ctgacaagtg ttnaggcaan    240 

attcctttct tgtgccc                                                   257 

 
           
             75  
             330  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(330)  
               n = A,T,C or G  
             
           
            75 

tgttcataag gctggtgata naggggtctt gtcatggaaa ggtgctcttc caggaaacct     60 

ctgtgtatgg aggtcgnagc cacaatacgc ggacgangat gtgaacacct acaatgccgc    120 

catcncttac accatcctca gccaagatcc tgagctccct gacnaaaata tgttcnccat    180 

taacaggaac gcaggagtca tcggtgtggt cnccactggg ctggaccgaa agagtttccc    240 

tacgtgtacc ntggtggttc aagcngctga ccttcanggt gaggggttaa tcacnacagc    300 

ancngctgtg atcacagtca ctgntaccaa                                     330 

 
           
             76  
             387  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(387)  
               n = A,T,C or G  
             
           
            76 

gctcgcgcgc ctgcaggtcg acactagtgg atccaaagaa ttcggcacga gaacaacagt     60 

tatctccaag atgctattcg ttgaacccat cctggaggtt tccagcttgc cgacaaccaa    120 

ctcaacaacc aattcagcca ccaaaataac agctaatacc actgatgaac ccaccacaca    180 

acccaccaca gagcccacca cccaacccac catccaaccc acccaaccaa ctacccagct    240 

cccaacagat tctcctaccc agcccactac tgggtccttc tgcccaggac ctgttactct    300 

ctgctctgac ttgganantc attcaacana agccgtgttg ggggaagctt tggtaaattt    360 

ctccctgaag ctctaccacg ccttctc                                        387 

 
           
             77  
             339  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(339)  
               n = A,T,C or G  
             
           
            77 

ctgctgatcn gggtcccttt ggagcacaga tgatgcnatg gccancnngg gacaacnacg     60 

tgatctgcgc cctggtcctg gtgtccatnc tggccctcgg nancctggcc gaggcccana    120 

canagacgtg tncagtggcc ccccgtgaaa gacagaattg tggttttcct ggtgtcacac    180 

cctcccantg tgcaaataag ggctgctgtt tcgacaacac cgttcgtggg gtcccctggt    240 

gcttctatcc taataccntc nacntcccnc canaaaagga ntgtgaattt tanacacttc    300 

tgcagggatc tgcctgcatc ctgacgcngt gccgtcccc                           339 

 
           
             78  
             385  
             DNA  
             Homo sapien  
           
            78 

tcggtcatag ggagagattt gtatgctgta ctatgcagcg tttaaagtta gtgggttttg     60 

tgatttttgt attgaatatt gctgtctgtt acaaagtcag ttaaaggtac gttttaatat    120 

ttaagttatt ctatcttgga gataaaatct gtatgtgcaa ttcaccggta ttaccagttt    180 

attatgtaaa caagagattt ggcatgacat gttctgtatg tttcagggaa aaatgtcttt    240 

aatgcttttt caagaactaa cacagttatt cctatactgg attttaggtc tctgaagaac    300 

tgctggtgtt taggaataag aatgtgcatg aagcctaaaa taccaagaaa gcttatactg    360 

aatttaagca aaaaaaaaaa acccc                                          385 

 
           
             79  
             307  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(307)  
               n = A,T,C or G  
             
           
            79 

tcgatacagg gatgtcagag ctgccagaga ctttatcctg aagctttacc aagatcagaa     60 

tcctgacaaa gnagaaagtc atctactctc acttcacatg tgctacagat acagacaata    120 

ttcgctttgt gtttgctgct gtcaaagaca caattctaca gctaanccta agggaattca    180 

accttgtcta aaagctgctg cccactcctc ccctataaca gaagatgtga tttgcaaact    240 

ccttgtttta tttgnaagtg cttctgacat cnccagagcc agccccatgc caggaactaa    300 

ggatgtc                                                              307 

 
           
             80  
             528  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(528)  
               n = A,T,C or G  
             
           
            80 

gtcgatacag gaacagcatg tccaaatcga tgtggatgtt tccaagcctg acctcacggc     60 

tgccctgcgt gacgtacgtc agcaatatga aagtgtggct gccaagaacc tgcaggaggc    120 

agaagaatgg tacaaatcca agtttgctga cctctctgag gctgccaacc ggaacaatga    180 

cgccctgcgc caggcaaagc aggagtccac tgagtaccgg agacaggtgc agtccctcac    240 

ctgtgaagtg gatgccctta aaggaaccaa tgagtccctg gaacgccaga tgcgttgaaa    300 

tggaagagaa ctttgccgtt gaagctgcta actaccaaga cactattggc cgcctgcagg    360 

atgagattca gaatatgaag ganggaaatg gctcgtcacc ttcgtgaata ccaagacctg    420 

ctcaatgtta agatggccct tgacattgaa attgccacct acanggaact gctggangcn    480 

aagaaaacca ggatttctct gcctcctccn aacttttcct cccctgaa                 528 

 
           
             81  
             369  
             DNA  
             Homo sapien  
           
            81 

agcatggctc ccgaagtttt gccaaaacct cggatgcgtg gccttctggc caggcgtctg     60 

cgaaatcata tggctgtagc attcgtgcta tccctggggg ttgcagcttt gtataagttt    120 

cgtgtggctg atcaaagaaa gaaggcatac gcagatttct acagaaacta cgatgtcatg    180 

aaagattttg aggagatgag gaaggctggt atctttcaga gtgtaaagta atcttggaat    240 

ataaagaatt tcttcaggtt gaattaccta gaagtttgtc actgacttgt gttcctgaac    300 

tatgacacat gaatatgtgg gctaagaaat agttcctctt gataaataaa caattaacaa    360 

aaaaaaaaa                                                            369 

 
           
             82  
             269  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(269)  
               n = A,T,C or G  
             
           
            82 

atgacaggga tgancaaact tngtctgggg tattgatgaa gatgacctac tgctgatgat     60 

accagtgctg ctgtaactga agaaatgcca ccccttgaag gagatgacga cacatcacgc    120 

atggaagaag tagactaatc tctggctgag ggatgactta cctgttcagt actctacaat    180 

tcctctgata atatattttc aaggatgttt ttctttattt ttgttaatat taaaangtct    240 

gtntggnatg acaactnctt taaggggaa                                      269 

 
           
             83  
             196  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(196)  
               n = A,T,C or G  
             
           
            83 

tttgggtcca attacagcta aagcaaaagt ggttattgaa ctgtttttat cggtctcggg     60 

nnttgctaaa ccttcccagg tgtattttgg aggtacagtt gttggcnagc aagctatnaa    120 

atctgaagat gaagtgggaa gttnaatana gtatgaatnc agggtaagaa actnaggtaa    180 

acctcnaata tncctc                                                    196 

 
           
             84  
             448  
             DNA  
             Homo sapien  
             
               misc_feature  
               (1)...(448)  
               n = A,T,C or G  
             
           
            84 

caaacatggg catggtgtca gcgataatgt ttntancagc tcccgacata aatcagtaan     60 

tnngatttcc accatatcna ncntcnggaa tttaaccntc aggagnagct cttnntcaga    120 

cnccctggaa aaacgagccc cattgnancc anctttgana cataaaacct ggagaaattc    180 

tccaatacng aaggtatana gcggggcatc gttgacagca tcacgggtca aaggcttctg    240 

gaggctcagg cctgcaaagg tggcatcatc cacccaacca cgggccagaa cctgtcnctt    300 

caggacgcag tctcccnggg tgtgattgac caagacatgg ccaccaggct gaagcctgct    360 

cagaaagcct tcataggctt cgagggtgtg aagggaaaga agaagatgtc agcagcagag    420 

gcagtgaaaa aaaaaaaacc cctatatt                                       448 

 
           
             85  
             169  
             DNA  
             Homo sapien  
           
            85 

agcagaccaa ctgccttttg tgagaccttc ccctccctat ccccaacttt aaaggtgtga     60 

gagtattagg aaacatgagc agcatatggc ttttgatcag tttttcagtg gcagcatcca    120 

atgaacaaga tcctacaagc tgtgcaggca aaacctagca ggaaaaaaa                169