Abstract:
A method of screening mammals for an autoimmune disease or a predisposition to said disease (e.g diabeties). The method consists of identifying polymorphisms in IL-12 p40 and linking them to the disease condition.

Description:
FIELD OF THE INVENTION  
         [0001]    The present invention relates generally to a method of screening mammalian animals for a disease condition or a predisposition for the development of a disease condition. More particularly, the present invention provides a method of screening for a disease condition or a predisposition for the development of a disease condition characterised by Th1/Th2 dysregulation. Disease conditions contemplated herein include autoimmune conditions such as, but not limited to, diabetes. The present invention is predicated in part on the determination of the presence of a particular form of IL-12 subunit or linkage between an IL-12 subunit and the disease condition.  
         BACKGROUND OF THE INVENTION  
         [0002]    Bibliographic details of the publications referred to by author in this specification are collected at the end of the description.  
           [0003]    The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the common general knowledge in Australia.  
           [0004]    Autoimmune diseases result from the body&#39;s immune system mounting an immune response to “self” via the aberrant activation of B cells and/or one or more of the subclasses of T cells. The classes of T cells can be defined broadly according to their requirement for particular molecules encoded by the major histocompatibility complex (MHC), and by their function. The class of T cells restricted by MHC class II molecules are generally referred to as “helper” T (referred to herein as “Th”) cells, and can be further divided into two main subclasses depending on the type of immune response which they mediate. These subclasses are referred to as Th1 and Th2, the former subclass mediating cellular immune response and the later mediating an antibody immune response.  
           [0005]    There is increasing evidence that some disease states, including autoimmune diseases, may result from dysregulation of Th1/Th2 status. Insulin dependent diabetes melitis (IDDM), for example, results from the dysregulation of T cells in that they may be mediated by an imbalance towards Th1 and Th2 type responses, respectively. Although a number of immunological influences which affect Th1 and Th2 responses have been identified, such as the influence of the cytokines interleukin-10 and interleukin-12 (herein referred to as “IL-12”), the precise molecular mechanisms of regulating the division of Th cells into these subclasses together with their functional regulation has not been elucidated.  
           [0006]    IL-12 is comprised of two subunits—p35 and p40. In work leading up to the present invention, the inventors have identified two allelic variants of the IL-12 p40 subunit. Analysis of distribution of these variants in the population has resulted in the surprising correlation of genetic variation in the IL-12 p40 genes with diseases having a bias in T cell response in terms of the Th subtype of the response. In accordance with the present invention, the inventors have identified a method of screening for an individual with a disease condition or predisposition for the development of a disease condition characterised by Th1/Th2 dysregulation. The developments described herein further facilitate the design of methodology to screen for individuals exhibiting resistance to the development of a disease condition characterised by Th1/Th2 dysregulation. In a further aspect there is now facilitated the development of methods of therapeutically and/or prophylactically treating such disease conditions.  
         SUMMARY OF HE INVENTION  
         [0007]    Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.  
           [0008]    The subject specification contains nucleotide sequence information prepared using the programme PatentIn Version 2.0, presented herein after the bibliography. Bach nucleotide sequence is identified in the sequence listing by the numeric indicator &lt;210&gt; followed by the sequence identifier (e.g. &lt;201&gt;1, &lt;210&gt;2, etc). The length, type of sequence (DNA, etc) and source organism for each nucleotide sequence is indicated by information provided in the numeric indicator fields &lt;211&gt;, &lt;212&gt; and &lt;213&gt;, respectively. Nucleotide sequences referred to in the specification are defined by the information provided in numeric indicator field &lt;400&gt; followed by the sequence identifier (e.g. &lt;400&gt;1, &lt;400&gt;2, etc).  
           [0009]    One aspect of the present invention provides a method of determining the presence of a disease condition or a predisposition for the development of a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
           [0010]    Another aspect of the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or other associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivatives thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
           [0011]    Still another aspect of the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
           [0012]    Yet another aspect of the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
           [0013]    Even more preferably said IL-12 p40 Taq1 +  allelic form comprises the nucleotide sequence substantially as set forth in&lt;400&gt;1.  
           [0014]    Still yet another aspect of the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
           [0015]    Yet still another aspect of the present invention provides a method of determining the presence of an autoimmune disease condition or a predisposition for the development of an autoimmune disease condition in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or a derivative thereof or its expression product is indicative of the presence of said autoimmune disease condition or the propensity to develop said autoimmune disease condition.  
           [0016]    A further aspect of the present invention provides a method of determining the presence of IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of said IDDM or the propensity to develop said IDDM.  
           [0017]    Another further aspect of the present invention provides a method of determining the presence of a disease condition or a predisposition for the development of a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
           [0018]    Yet another further aspect of the present invention provides a method of determining IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
           [0019]    Still another further aspect of the present invention provides a method of determining the presence of IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said Taq1 − allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
           [0020]    Yet still another further aspect of the present invention there is provided a method of determining resistance to a disease condition in a mammal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of resistance to developing said disease condition.  
           [0021]    Still yet another further aspect of the present invention provides a method of determining resistance to a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or derivative thereof is indicative of a resistance to developing said disease condition.  
           [0022]    Another aspect of the present invention provides a method of determining resistance to IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of a resistance to developing IDDM.  
           [0023]    Yet another aspect of the present invention there is provided a method of determining resistance to a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
           [0024]    Still another aspect of the present invention provides a method of determining resistance to IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
           [0025]    The present invention should also be understood to extend to methods of detecting novel IL12 p40 polymorphisms based on the use of familial gene transfer linkage studies.  
           [0026]    A kit for determining the presence of a disease condition or a predisposition to the development of a disease condition in a mammalian animal said kit comprising a means of detecting the presence or absence of a form of IL-12p40 genetic sequence or derivative thereof or its expression product.  
           [0027]    Another aspect of the present invention provides a kit for determining the presence of a disease condition or a predisposition to the development of a disease condition in a mammalian animal said kit comprising in compartmental form a first compartment adapted to contain an agent for detecting the form of IL-12 p40 genetic sequence or derivative thereof or its expression product and a second compartment adapted to contain reagents useful for facilitating the detection by the agent in the first compartment. Further compartments may also be included, for example, to receive a biological sample. The agent may be an oligonucleotide or antibody or other suitable detecting molecule.  
           [0028]    Yet another aspect of the present invention provides a kit for determining resistance to a disease condition in a mammalian animal said kit comprising in compartmental form a first compartment adapted to contain an agent for detecting the form of IL-12 p40 genetic sequence or derivative thereof or its expression product and a second compartment adapted to contain reagents useful for facilitating the detection by the agent in the first compartment. Further compartments may also be included, for example, to receive a biological sample. The agent may be an oligonucleotide or antibody or other suitable detecting molecule.  
           [0029]    The present invention further contemplates a method of treatment and/or prophylaxis of the disease conditions herein defined said method comprising administering to a mammal an effective amount of a form of IL-12 p40 genetic sequence or derivative, agonist or antagonist thereof or a molecule which regulates the functioning of said IL-12 p40 genetic sequence or its expression product or derivative, antagonist or agonist thereof wherein said IL-12 p40 or regulatory molecule thereof promotes resistance to said disease condition.  
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0030]    [0030]FIG. 1 is a schematic representation of a high resolution map of the IL12p40 locus.  
         [0031]    A. Placement of IL-12p40 on the radiation hybrid map of chromosome 5q33 relative to the genes GABRA1 (Johnson et al., 1992) and GABRA6 (Hicks et al., 1994) and microsatellite markers (Weissenbach et al., 1992). Oligonucleotides were designed to amplify sequences from the 3′ untranslated region of the human IL-12p40 gene but not from hamster genomic DNA. Radiation hybrids from the Genebridge 4 series (Research Genetics, AL) was tested for human IL-12p40. Additional primers, including GABR41AA (Johnson et al., 1992) and D5S403, D5S410 and D5S412 (Weissenbach et al., 1992) were also tested. The results were used to search against the Whitehead Institute database (http://wwwgenome.wi.mit.edu/cgi-bin/contig/rhmapper.pl) using a LOD=15 for linkage to the framework map.  
         [0032]    B. Detailed restriction maps of the PAC and BAC clones containing IL-12p40. A PAC containing IL12p40 was isolated by screening pools from the human PAC library produced by (Ioannou et al., 1994). The direction of transcription of the gene is shown by the arrow. The marker 93/SP6 was obtained from the end sequence of PAC93-1, and used to screen a BAC library. The resulting clone, BAC 626-19, had an 165 kb insert containing the entire PAC93-1 insert (130 kb) with an additional 2.5- and 30-kb at its SP6 and T7 ends, respectively. Restriction enzyme maps were prepared after digestion with NotI, SalI, SacII and MluI, followed by resolution by pulsed field gel electrophoresis, and hybridization with oligonucleotides complementary to the vector ends (T7 or SP6) or to the promoter or 3UTR of IL-12p40.  
         [0033]    C. Genomic organisation of the human IL-12p40 gene By comparing the complete genomic sequence with published cDNA sequences, the position of exons 1-8 and introns was deduced. Open boxes=coding exons; first and last exons are non-coding. Size of introns is indicated below the line. Start and stop codons are indicated. The asterisk indicates the presence of a mRNA degradation motif (Zubiaga et al., 1995). Arrows indicate approximate positions of confirmed polymorphisms (see Tables 8, 9 and 10).  
         [0034]    [0034]FIG. 2 is a schematic representation of the Complete genomic sequence of the IL-12p40 gene (&lt;400&gt;130). The sequence starts 2,397 nucleotides upstream of the TATA box and overlaps the previously published partial promoter sequence. The eight exon sequences (underlined) were determined by comparison with the IL-12p40 cDNA sequences. The translation initiation (ATG) and termination (TAG) codons are double underlined. The 9 base AU-rich element (ARE) consensus sequence is indicated by thick underlining.  
         [0035]    [0035]FIG. 3 is a graphical representation of linkage of TID to chromosome 5q. Families with at least two affected sibs were genotyped at markers extending over 33 cM of chromosome 5q. Multipoint linkage analysis was undertaken using the MAPMAKER/Sibs software program (Kruglyak et al., 1995). Output shows maximized lod scores (Holmans P., 1993) for all 249 sibpairs from 187 multiplex families (dashed line). MLS scores were also determined for sibpairs who were either identical (HLA IBD) or mismatched (HLA MIS). Dotted line indicates MLS=2.3, which may be taken as significant evidence for linkage in a single test for linkage (Holians P., 1993). Markers were microsatellite repeats (Weissenbach et al., 1992) including the highly polymorphic repeat within the gene GABRA1.  
         [0036]    [0036]FIG. 4 is a diagramatic representation of TDT of IL12B markers placed on the physical map of 5q33-34.  
         [0037]    A. Physical map of YAC (left) and BAC (right) clones containing IL12B. The transcriptional orientation is shown with respect to the centromere. The location of the D5S2937 TAA repeat (box) and the SP6 end (circle) of the PAC clone 93.1 is shown in relation to the genetic markers within the promoter, intron 4 and 3′ UTR of IL12B. Additional anonymous markers on the YAC map are indicated as crosses; the ADRA1b is located telomeric to, and is in the same transcriptional orientation as, IL12B. YAC&#39;s shown are (from left): 917b7, 910b3 and 756f1. Further YAC and PAC details were described previously (Huang et al., in press). Restriction sites were determined for Notl (Not), Sacl (Sac), M7ul (Mlu) and Sall (Sal).  
         [0038]    B. TDT of IL12B markers. Results from families in which sibs show linkage to IL12B (that is, “IBD 2”) or not (“IBD 1/0”) are shown as the negative log of the P value returned from the TDT. IBD status was assessed by genotypes at the highly polymorphic nearby GABRA4 locus and also at flanking markers. Transmission ratio of the 3′ UTR alleles in IBD 2 families was 76:33 and in IBD 1/0 families was 95:89; of the intron 4 alleles was 61:22 and 76:63; and of the promoter alleles was 44:61 and 132:130, respectively.  
         [0039]    [0039]FIG. 5 is a diagramatic representation of allele-dependent expression of IL12B.  
         [0040]    A. Total RNA was isolated from 1/1 and 2/2 EBV-transformed cell lines and northern analysis was performed with human IL12B and GAPDH cDNA probes.  
         [0041]    B. The levels of IL12B MRNA in each cell line relative to GAPDH was determined by densitometry. Bars show mean±s.e. for three separate experiments.  
         [0042]    [0042]FIG. 6 is a schematic representation of the sequence determination and comparison of IL12p40 promoter alleles in humans.  
         [0043]    A. The sequence of the region containing the polymorphism 5′ of the IL-12p40 gene is shown (&lt;400&gt;131). This sequence was determined from a PAC clone we isolated. Underlined=match with published seq (GENBANK HSU89323, Ma et al); bold=oligos used to amplify polymorphism. NB; we have designed another REV oligo to allow for generation of a smaller PCR product. (The first rev oligo is indicated by italics).  
         [0044]    B. Alignment of human alleles 1 (&lt;400&gt;132) and 2 (&lt;400&gt;133) showing the complex change involving the insertion of 5 bases and the deletion of a G resulting in a nett gain of 4 bases compared to the shorter allele 2. No other differences were found in a total of 2 kb sequenced upstream of the IL-12p40 gene.  
     
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
       [0045]    The present invention is predicated, in part, on the identification of allelic variants of an IL 12 subunit and more particularly p40 subunit of IL-12 and the surprising observation that a correlation exists between the expression of a particular genetic variant and the onset of an autoimmune disease condition such as, but not limited to, IDDM. Although not intending to limit the invention to any one theory or mode of action, it is proposed that genetic variation in the IL-12 p40 gene modulates the expression levels of the RNA thereby modulating the levels of the IL-12 protein and thereby biasing the Th cell response either towards a Th2 type response or a Th1 type response. This proposed mechanism of action now provides a means for the development of a method of screening individuals to determine a predisposition to developing diseases involving the dysregulation of the Th1/Th2 response and or resistance thereto a means for the rational design of therapeutic or prophylactic regimes and/or molecules for modulation of the Th1/Th2 response.  
         [0046]    Accordingly, one aspect of the present invention provides a method of determining the presence of a disease condition or a predisposition for the development of a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
         [0047]    More particularly the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or other associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivatives thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of the disease condition or the propensity to develop said disease condition.  
         [0048]    The term “mammalian animal” includes humans, primates, livestock animals (e.g. horses, cattle, sheep, pigs, donkeys) laboratory test animals (e.g. mice, rats, rabbits, guinea pigs) companion animals (e.g. dogs, cats) and captive wild animals (e.g. kangaroos, deer, foxes). Preferably, the mammal is a human or a laboratory test animal. Even more preferably the mammal is a human.  
         [0049]    IL-12 is a heterodimeric glycoprotein composed of unrelated subunits of 35 kDa (p35) and 40 kDa (p40). Accordingly, reference to “IL-12 p40 genetic sequence” should be understood as a reference to all forms of DNA- and RNA encoding (i) all or part of the p40 subunit of IL-12 and derivatives thereof or (ii) all or part of a regulatory sequence (such as a promoter sequence) which directly or indirectly regulates the expression of the IL-12 p40 subunit and is located at a position other than between the IL-12 p40 genomic DNA transcription initiation and termination sites and derivatives thereof.  
         [0050]    This definition includes, but is not limited to, all forms of the IL-12 p40 genomic DNA sequence, for example:  
         [0051]    (i) allelic variants such as the Taq1 +  (&lt;400&gt;1) and Taq1 −  (&lt;400&gt;2), allelic forms which are defined on the basis of the presence of a deoxycytosine or deoxyadenine nucleotide, respectively, at position 235 of &lt;400&gt;1 and &lt;400&gt;2. &lt;400&gt;1 and &lt;400&gt;2 are partial IL-12 p40 cDNA sequences and depict the 3′ end of the IL-12 p40 cDNA. Position 235 occurs in the 3′ untranslated region of the cDNA sequence.  
         [0052]    (ii) allelic variants such as those characterised by promotor region polymorphisms (&lt;400&gt;3, &lt;400&gt;4, &lt;400&gt;5, &lt;400&gt;6, &lt;400&gt;7, &lt;400&gt;8, &lt;400&gt;132, &lt;400&gt;133).  
         [0053]    (iii) allelic variants such as those characterised by polymorphisms in exon 6 (&lt;400&gt;9, &lt;400&gt;10), exon 7 (&lt;400&gt;11, &lt;400&gt;12) or exon 8 (&lt;400&gt;13, &lt;400&gt;14).  
         [0054]    (iv) allelic variants such as those characterised by polymorphisms in intron 1 (&lt;400&gt;41&lt;400&gt;48), intron 2 (&lt;400&gt;49-&lt;400&gt;52), intron 4 (&lt;400&gt;55, &lt;400&gt;58) and intron 7 (&lt;400&gt;59, &lt;400&gt;60).  
         [0055]    (v) allelic variants characterised by the presence of any one or more of the polymorphisms detailed in (i)-(iv) all forms of the RNA transcribed from said IL-12 p40 genomic DNA sequence (for example the primary RNA transcript, mRNA or splice variants of the RNA transcript) and the cDNA generated from RNA transcribed from said IL-12 p40 genetic sequence.  
         [0056]    Preferably said IL-12 p 4 0 is the Taq1 +  and/or Taq1 −  allelic form.  
         [0057]    According to this preferred embodiment the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of a propensity to develop said disease condition.  
         [0058]    More preferably the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of a propensity to develop said disease condition.  
         [0059]    Even more preferably said IL-12 p40 Taq1 +  allelic form comprises the nucleotide sequence substantially as set forth in&lt;400&gt;1.  
         [0060]    In another preferred embodiment the present invention provides a method of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation or a predisposition for the development of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of a propensity to develop said disease condition.  
         [0061]    Even more preferably said IL-12 p40 Taq1 −  allelic form comprises the nucleic acid sequence substantially as set forth in&lt;400&gt;2.  
         [0062]    It should be understood that the presence of the Taq1 +  or Taq1 −  polymorphism may be indicative of a number of disease conditions characterised by Th1/Th2 dysregulation. In one embodiment, to the extent that the disease condition is IDDM, Taq1 −  expression in an individual is indicative of a propensity to develop IDDM while Taq+expression in an individual is indicative of resistance to the development of IDDM.  
         [0063]    Reference to “expression product” should be understood as a reference to the peptide, polypeptide or protein resulting from the translation of IL-12 p40 RNA sequences or transcription and translation of IL-12 p40 DNA sequences as hereinbefore defined.  
         [0064]    Reference to a disease condition “characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation” should be understood as a reference to a disease condition in which at least some of the pathology associated with said disease condition is either directly or indirectly due to the activation of a particular subpopulation of Th cells. For example, IDDM is characterised by a Th1 type response. Preferably, said disease condition is an autoimmune disease condition.  
         [0065]    Accordingly, another aspect of the present invention provides a method of determining the presence of an autoimmune disease condition or a predisposition for the development of an autoimmune disease condition in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or a derivative thereof or its expression product is indicative of the presence of said autoimmune disease or the propensity to develop said autoimmune disease condition.  
         [0066]    Preferably, said autoimmune disease condition is an autoimmune disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation.  
         [0067]    Even more preferably, said allelic form of IL-12 p40 is the Taq1 +  or Taq1 −  form.  
         [0068]    Without limiting the present invention to any one theory or mode of action, disease conditions characterised by Th1/Th2 dysregulation are thought to be mediated by an imbalance in the Th response in that it is incorrectly skewed towards either a Th1 or Th2 response. The skewing of Th cells towards either a Th1 or a Th2 response is now envisaged as at least partly regulated by genetic variation in the IL-12 p40 gene which acts to modulate the expression levels of the IL-12 p40 polypeptide. Analysis of the frequency of Taq1 +  and Taq1 −  IL-12 p40 alleles in subjects who exhibit symptoms of IDDM indicates that expression of a Taq1 −  allele is indicative of susceptibility to IDDM while expression of a Taq1 +  allele is indicative of resistance to IDDM.  
         [0069]    According to this most preferred embodiment, the present invention provides a method of determining the presence of IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 −  allelic form of 1L-12 p40 genetic sequence or derivative thereof or its expression product is indicative of the presence of said IDDM or the propensity to develop said IDDM.  
         [0070]    The present invention should be understood to extend to methods of determining the presence of a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation, or a predisposition thereof, by screening for the combination of a Taq1 + /Taq1 −  polymorphism together with any other polymorphism expressed by an individual.  
         [0071]    Still without limiting the invention to any one theory or mode of action, transmission disequilibrium studies have further indicated that in certain disease conditions characterised by Th1/Th2 dysregulation the Taq1 +  and Taq1 −  allelic forms of IL-12 p40 are indicative of a predisposition to developing said disease when they have been transmitted to the affected mammal in a form where they are linked to another gene. The method of the present invention is exemplified herein utilising the genetic marker GABRAL which occurs in two allelic forms—A and B. Expression of the Taq1 − /GABRA1-A haplotype where the two genes are linked is indicative of IDDM susceptibility while expression of the Taq1 − /GABRA1-A haplotype where the two genes are unlinked is not indicative of IDDM susceptibility. Conversely, transmission of the Taq1 + /GABRA-A haplotype in a linked form is indicative of IDDM resistance.  
         [0072]    Accordingly, a related aspect of the present invention provides a method of determining the presence of a disease condition or a predisposition for the development of a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
         [0073]    Reference to genes being “linked” is a reference to any two or more genes which do not assort independently at meiosis. Determining the linkage of two genes can be achieved by any one of a number of methods including for example screening one or more parents of said mammal to determine the pattern of gene transmission and thereby the degree of linkage between the IL-12 p40 gene or derivative thereof and another gene. Alternatively, said linkage can be determined by screening one or more parents and comparing with a proband.  
         [0074]    Preferably said form of IL-12 p40 is an allelic form of IL-12 p40.  
         [0075]    In a most preferred embodiment, said disease condition is an autoimmune disease condition and most preferably IDDM.  
         [0076]    According to this most preferred embodiment the present invention provides a method of determining IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
         [0077]    Most preferably the present invention provides a method of determining the presence of IDDM or a predisposition for the development of IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said Taq1 −  allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
         [0078]    Preferably, the gene to which said IL-12 p40 genetic sequence is linked is an informative genetic marker. By “informative” it is meant a genetic marker which when used in conjunction with said IL-12 p40 genetic sequence improves or otherwise indicates involvement of said IL-12 p40 genetic sequence in the subject disease or other condition. Preferably, said informative genetic marker is a GABRA allele.  
         [0079]    Even more preferably, said other gene is the GABRA1-A allele genetic marker.  
         [0080]    The expression of a particular form of IL-12 p40 is also indicative of a mammal&#39;s resistance to developing a disease condition characterised by Th1/Th2 dysregulation.  
         [0081]    Accordingly, in another aspect of the present invention there is provided a method of determining resistance to a disease condition in a mammal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of resistance to developing said disease condition.  
         [0082]    Preferably, said disease condition is characterised, exacerbated or otherwise associated with Th1/T2 dysregulation.  
         [0083]    More preferably said form of IL-12.p40 genetic sequence is an allelic form of IL-12 p40 genetic sequence.  
         [0084]    According to this most preferred embodiment the present invention provides a method of determining resistance to a disease condition characterised, exacerbated or otherwise associated with Th1/Th2 dysregulation in a mammalian animal said method comprising screening for the presence of an allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said allelic form of IL-12 p40 genetic sequence or derivative thereof is indicative of a resistance to developing said disease condition.  
         [0085]    Most preferably said allelic form of IL-12 p40 is the Taq1 +  or Taq1 −  allelic form.  
         [0086]    In a most preferred embodiment said disease condition is an autoimmune disease condition and even more preferably IDDM.  
         [0087]    According to this most preferred embodiment the present invention provides a method of determining resistance to IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein the presence of said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product is indicative of a resistance to developing IDDM.  
         [0088]    In another related aspect of the present invention there is provided a method of determining resistance to a disease condition in a mammalian animal said method comprising screening for the presence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
         [0089]    Preferably, said disease condition is characterised, exacerbated or otherwise associated with Th1[Fh2 dysregulation. Even more preferably said disease condition is an autoimmune disease condition and most preferably IDDM.  
         [0090]    Still more preferably said IL-12 p40 is the Taq1 +  allelic form.  
         [0091]    According to this most preferred embodiment the present invention provides a method of determining resistance to IDDM in a mammalian animal said method comprising screening for the presence of the Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof or its expression product wherein said Taq1 +  allelic form of IL-12 p40 genetic sequence or derivative thereof is linked to another gene.  
         [0092]    Most preferably said other gene is GABRA1-A allele genetic marker.  
         [0093]    Reference to detecting “resistance” should be understood to generally refer to detecting a reduction in the pathology associated with an existing disease condition, preventing, delaying or minimising the onset of pathology associated with the onset of said disease condition, or preventing the onset of said disease condition.  
         [0094]    The present invention should also be understood to extend to methods of detecting novel IL-12 p40 polymorphisms based on the use of familial gene transfer linkage studies. Further sequence polymorphisms may exist in the vicinity of the IL-12 p40 gene. Some of these may also be involved in regulating IL12 p40 gene expression and therefore the ability to produce Th1 or Th2 dominated immune response and hence resistance or susceptibility to autoimmune disease. Such polymorphisms may be tested by their co-segregation with the IL-12 p40 Taq− allele to IDDM subjects, or by their non-transmission in linkage with the IL12 p40 Taq+ allele. An example of how such an additional polymorphism may be detected and its utility is provided with reference to the GABRA-A genotype in Table 6. Such additional polymorphisms may be tested in, for example, functional assays by in vitro transfection experiments using appropriate reporter constructs.  
         [0095]    Screening of the forms of IL-12 p40 genetic sequences or derivatives thereof or its expression products may be achieved utilizing any of a number of techniques including PCR analysis and antibody binding assays.  
         [0096]    In one preferred method, the IL-12 p40 gene or transcribed RNA is subjected to PCR or RT-PCR, respectively, using primers homologous to gene sequences located 5′ and 3′ of the Taq1 polymorphism. The oligonucleotide is generally labelled with a reporter molecule capable of giving an identifiable signal such as a radioisotope, chemiluminesce molecule or a fluorescent molecule. A particularly useful reporter molecule is a biotinylated molecule.  
         [0097]    Another useful detection system involves antibodies directed to the various forms of IL-12 p40 genetic sequences, to the Taq1 polymorphism itself or to the expression products of the various IL-12 p40 forms. Detection utilising antibodies may be accomplished immunologically in a number of ways such as by Western Blotting and ELISA procedures. These procedures include both single site and two site or “sandwich” assays of the noncompetitive type, as well as the traditional competitive binding assays. These assays also include direct binding of a labelled antibody to a target.  
         [0098]    Another aspect of the present invention provides a kit for determining the presence of a disease condition or a predisposition to the development of a disease condition in a mammalian animal said kit comprising a means of detecting the presence or absence of a form of IL-12 p40 genetic sequence or derivative thereof or its expression product.  
         [0099]    Without limiting this aspect of the present invention in any way, the subject kit may be designed to detect either the presence of a given allele, or its absence, in an individual. In a preferred embodiment the presence of a specific allele is screened for. The means by which the subject kit detects the form of IL-12 p40 may be any suitable means including, but not limited to, any mass spectrometry technique, gels, DNA or protein chips, DNA probing means, antibody or other immunological reagent.  
         [0100]    In one embodiment the present invention provides a kit for determining the presence of a disease condition or a predisposition to the development of a disease condition in a mammalian animal said kit comprising in compartmental form a first compartment adapted to contain an agent for detecting the form of IL-12 p40 genetic sequence or derivative thereof or its expression product and a second compartment adapted to contain reagents useful for facilitating the detection by the agent in the first compartment. Further compartments may also be included, for example, to receive a biological sample. The agent may be an oligonucleotide or antibody or other suitable detecting molecule.  
         [0101]    In another embodiment the present invention provides a kit for determining resistance to a disease condition in a mammalian animal said kit comprising in compartmental form a first compartment adapted to contain an agent for detecting the form of IL-12 p40 genetic sequence or derivative thereof or its expression product and a second compartment adapted to contain reagents useful for facilitating the detection by the agent in the first compartment. Further compartments may also be included, for example, to receive a biological sample. The agent may be an oligonucleotide or antibody or other suitable detecting molecule.  
         [0102]    The present invention further contemplates a method of treatment and/or prophylaxis of the disease conditions hereinbefore defined said method comprising administering to a mammal an effective amount of a form of IL-12 p40 genetic sequence or derivative, agonist or antagonist thereof or its expression product or derivative, antagonist or agonist thereof wherein said IL-12 p40 promotes resistance to said disease condition. For example, in patients suffering from IDDM or a predisposition to developing IDDM the Taq1 +  form of the IL-12 p40 gene or transcription or translation product or molecules which regulate Taq1 +  functioning or expression may be administered. The present invention facilitates modulation of the immune system response both in disease states or in non-disease states where it is nevertheless desirable (for example, to regulate IL-12 levels as part of a vaccination protocol).  
         [0103]    Administration of said IL-12 p40 can be achieved via one of several techniques including, but in no way limited to:  
         [0104]    (i) Introduction of a nucleic acid molecule encoding a particular form of IL-12 p40 or a derivative thereof to modulate the capacity of that cell to synthesize said IL-12 p40.  
         [0105]    (ii) Introduction into a cell of a proteinaceous IL-12 p40 molecule of particular form or derivative thereof.  
         [0106]    The present invention may be used for the screening of individuals, families and populations. In this regard, the inventors have determined that the relationship between Taq1 allele expression and IDDM resistance or susceptibility is particularly evident in individuals who are ethnically of Northern European or United Kingdom origin. Accordingly, in a preferred embodiment the methods of the present invention are directed to screening individuals of this ethnic origin.  
         [0107]    Further features of the present invention are more fully described in the following nonlimiting Examples. It is to be understood, however, that this detailed description is included solely for the purposes of exemplifying the present invention. It should not be understood in any way as a restriction on the broad description of the invention as set out above.  
       EXAMPLE 1  
     Detection of IL-12 Taq Polymorphism  
       [0108]    A polymorphism was found in the 3′ UT region of the IL-12 p40 gene. This polymorphism was detected as follows. DNA was obtained from peripheral blood lymphocytes using standard techniques, and used to initiate polymerase chain reaction (PCR) using synthetic oligonucleotides and Taq DNA polymerase (Gibco). The sequences of these oligos were as follows:  
                                               FORWARD   TAGCTCATCTTGGAGCGAAT   (&lt;400&gt;134)                           REVERSE   AACATTCCATACATCCTGGC   (&lt;400&gt;135)          
 
         [0109]    Reverse oligo hybridises to the following sequence in 3′ UT region:  
                                           GCCAGGATGTATGGAATGTT   (&lt;400&gt;136)              
 
         [0110]    Following PCR, aliquots of the reaction products were incubated with Taq1 restriction enzyme (Promega) under conditions suggested by the manufacturer. The samples were then ran on gels (either agarose gels, or acrylamide gels if the primer was first labelled with 32p ATP) to determine the lengths of the DNA fragments. Using the above primers, a product of approximately 0.3 kbp is generated; if the Taq1 site is present, this yields fragments of approximately 0.14 and 0.16 kbp after digestion. Allele 1 is designated as the allele not digested by Taq1; allele 2 contains the Taq1 site (i.e. TCGA).  
         [0111]    Other methods for detecting this polymorphism include use of different oligonucleotides flanking the Taq1 site; use of allele-specific primers to preferentially amplify allele 1 (Taq1 −  polymorphism) or allele 2 (Taq1 +  polymorphism) sequences; testing products by hybridisation using allele-specific oligonucleotides; testing products or fragments derived therefrom for differences in mass by appropriate methods, e.g. mass spectrometry.  
       EXAMPLE 2  
     Determining the Frequency of Alleles 1 and 2 in Control Subjects  
       [0112]    The frequency of alleles 1 and 2 in controls was determined by typing DNA samples from anonymous donors.  
       EXAMPLE 3  
       [0113]    IL-12 Allele Expression and IDDM Susceptibility  
         [0114]    The role of IL-12 p40 alleles in insulin-dependent diabetes mellitus (IDDM) was tested by determining whether either allele was preferentially transmitted to affected offspring.  
         [0115]    These alleles were typed as described in Example 2. Transmission or nontransmission of these alleles from appropriate parents to affected offspring was determined using the Transmission Disequilibrium test (TDT) in the Genetic Analysis System programs (A. Young, GAS Manual User Guide v1.2 (Oxford University, 1995). Results for families are shown in table 1.  
                                                           TABLE 1                           Transmission of IL-12 p40 alleles in IDDM families                Allele                       Both parents   Trans   Not   Prob (binom)                            1   159   110   0.0017           2   110   159                                  
 
         [0116]    These results show that allele 1 is preferentially transmitted and allele 2 is preferentially not transmitted to IDDM offspring.  
       EXAMPLE 4  
       [0117]    Confirmation of the use of the Taq allele as an indicator of susceptibility to IDDM was obtained from an independent sample of 238 families recruited through the Royal Melbourne and Royal Children&#39;s Hospitals.  
                                                         TABLE 2                                   Allele   Trans   Not Trans   p                                        1   58   32   0.004           2   32   58   1                      
 
         [0118]    These data indicate that allele 1 is preferentially transmitted (i.e. confers susceptibility or is in linkage disequilibrium with the polymorphism that confers susceptibility) and that allele 2 is preferentially not transmitted (i.e. confers resistance).  
         [0119]    When these data are analysed with respect to ethnic origin, the following is found:  
                                                           TABLE 3                           North European ethnicity                Allele   Trans   Not Trans   p                            1   49   18   9.7e−05 +++           2   18   49   1                      
 
         [0120]    [0120]                                                           TABLE 4                           Non-North European ethnicity                Allele   Trans   Not Trans   p                            1   9   14   0.8           2   14   9   0.2                        
         [0121]    This suggests that this gene is particularly indicative of fDDM in individuals who ethnically originate from the UK and northern Europe.  
         [0122]    Pooling the Example 3 data together with the Example 4 data indicates that the total p value in all families (i.e. unselected for linkage to GABRA) is 6×10 −6 .  
       EXAMPLE 5  
     Linked IL-12 Allele Transmission and IDDM Susceptibility  
       [0123]    A linked genetic marker in the GABRA-A receptor α1-subunit gene (GABRA1) was also typed. The GABRAL alleles were detected as described by Johnson, K J, et al  Genomics  14:745-8. These alleles were subsequently simplified for the transmission disequilibrium analysis as they were found to fall into two distinct groups: the six highest MW alleles were designated as “A” and the three lowest were designated as “B”.  
         [0124]    DNA was analysed from families in which at least two children had IDDM. Linkage was assessed by evaluating whether the affected sibs had inherited GABRA1 and linked genes identical-by-descent (IBD) i.e. whether they had inherited the same maternal and the same paternal alleles at GABRA1 and/or at other flanking markers. IBD status for a particular chromosomal region suggests that affected sibs share gene(s) which influence disease susceptibility; such sibs are said to show genetic linkage to the markers shared IBD). Two groups could thus be defined: those who were IBD at the IL-12 p40/GABRA region, and those who were not IBD. Transmission of alleles 1 and 2 were evaluated in sibs showing linkage to GABRA1/IL-12 p40. Families showing no linkage to IL-12 p40 did not show preferential transmission of either allele. Families whose affected sibs showed linkage to IL-12 p40/GABRA1 showed preferential transmission of allele 1, and preferential non-transmission of the other allele. This indicated that these alleles were associated with IDDM susceptibility and resistance, respectively (Table 5).  
                                                           TABLE 5                           Transmission of IL-12 p40 alleles in sibs       showing linkage to IL-12 p40.                Allele                       Both Parents   Trans   Not   Prob (binom)                            1   102   61   0.00082           2   61   102                                  
 
         [0125]    By considering the IL-12 alleles and the linked GABRA1 marker, four haplotypes were defined, as follows. Haplotype 1A, IL-12 p40 allele 1, GABRA1 A; haplotype 1B, IL-12 p40 allele 1, GABRAL B; haplotype 2A, IL-12 p40 allele 2, GABRA1 A; haplotype 2B, IL-12 p40 allele 2, GABRA1 B. Transmission or nontransmission of these haplotypes from appropriate parents to affected offspring was determined using the Transmission Disequilibrium test (IDT) in the Genetic Analysis System programs (A. Young, GAS Manual User Guide v1.2 (Oxford University, 1995).  
         [0126]    Families showing no linkage to IL-12 p40 did not show preferential transmission of any haplotype. Families whose affected sibs showed linkage to IL-12 p40/GABRA1 (suggesting that IL-12 p40 may be contributing to their development of IDDM) showed preferential transmission of one haplotype, and preferential non-transmission of another haplotype. This indicated that these haplotypes were associated with IDDM susceptibility and resistance, respectively. (Table 6). Two other haplotypes showed no deviation in transmission, suggesting that they were neutral in conferring susceptibility.  
                                                                   TABLE 6                           Transmission Disequilibrium Test of IL-12 haplotypes in IDDM.                Linkage                           Status   Haplotype   Trans   Not Trans   P                            Unlinked   1A   84   80   —               1B   74   67   —               2A   22   27   —               2B   25   29   —           Linked   1A   94   52   0.00032                1B   62   66   —               2A   8   34   0.000006               2B   24   36   —                                  
 
       EXAMPLE 6  
     Complete Primary Structure, Chromosomal Localisation and Definition of Polymorphisms of the Gene Encoding the Human IL-12p40 Subunit  
       [0127]    High Resolution Mapping  
         [0128]    Although IL-12p40 had been mapped to chromosome 5q31-33 (Wairington et al., 1994), its position relative to microsatellite markers used in genetic studies has not been reported. Therefore, to localize IL-12p40 relative to other genes (eg GABRA1 (Johnson et al., 1992) and GABRA6 (Hicks et al., 1994)) and genetic markers D5S403, D5S10 and D5S412 9 in this region, radiation hybrid mapping was used (Boehnke et al., 1991). Comparison with previously mapped markers confirmed the assignment of IL-12p40 to distal chromosome 5q FIG. 1A). The optimal location for this gene was 3.3 centiRays (cR) telomeric from the microsatellite marker D5S412, and 3 cR centromeric from the anonymous DNA sequence, WI-9929 and a further 2.2 cR from D5S403 (FIG. 1A). These results integrating the genetic (Weissenbach et al., 1992) and physical maps of distal chromosome 5q will be useful for further genetic studies examining potential roles for IL-12p40 in disease. An inspection of the map of distal human chromosome 5 does not reveal any known diseases mapping to this region which could be attributable to variants in IL-12p40.  
         [0129]    A phage P1-derived artificial chromosome (PAC) clone was isolated from the human PAC library produced by (Ioannou et al., 1994) with primers designed to amplify a segment from the 3′ untranslated region of IL12p40. The PAC clone 93-1 had an insert size of 130 kb. The sequence from the SP6 end of this clone was used to design primers to isolate an overlapping clone from a bacterial artificial chromosome (BAC) library (Osoegawa et al., 1998). A high resolution map of these clones is shown in FIG. 1B. Characterisation of these clones showed that they both contained the complete gene which also was arranged 3′-&gt;5′ with respect to the centromere. The MluI site within the IL-12p40 promoter was located 90 kb from the SP6 end of the PAC clone (FIG. 1B).  
         [0130]    Determination of IL-12p40 Genomic Sequence  
         [0131]    Only part of the promoter and the exon 1 genomic sequences of human IL-12p40 have been determined previously (Ma et al., 1996). In order to complete the sequence of the human IL-12p40 gene, the PAC clone, 93-1, was used as a template. DNA sequencing was performed using a walking strategy with fluorochrome-labeled dideoxynucleotide terminators, followed by automated analysis. This strategy was used because the priming oligonucleotides could also be used to generate PCR products for polymorphism testing.  
         [0132]    A total of over 18 kb of sequence was determined and is deposited with Genbank (FIG. 2 provides the complete genomic sequence of the IL-12p40 gene). By alignment of this genomic sequence with the previously published cDNA sequences (Gubler et al., 1991; Wolf et al., 1991) the exact location and sequences of the exons were defined. Sequences at the exon-intron boundaries are reported in Table 7. The organization of the IL-12p40 gene is shown schematically in FIG. 1C. An unusual feature of the gene is that it has untranslated exons at both its 5′ and 3′ ends. Translation from its corresponding mRNA would be initiated at the first codon in exon 2 and would terminate at the last codon of exon 7.  
         [0133]    Comparison of Genomic and cDNA Sequences  
         [0134]    Sequence differences were observed between the sequence of the PAC clone that was determined and the previously reported IL-12p40 sequences, as shown in Table 8. These sequence differences could either represent genetic polymorphisms or arise from sequencing errors. In order to test whether the differences between the PAC sequence and the cDNA sequences were representative of alleles of the IL-12p40 gene, primers were designed to amplify the relevant regions of the gene from genomic DNA of different individuals (anonymous donors of European descent). PCR products were tested for the presence of genetic variants by either restriction enzyme digestion (where appropriate) or by direct sequencing. In this way, the A-&gt;C change in the 3UIR, resulting in creation of a TaqI site, was defined as a true genetic variant. In contrast, the C-&gt;G change resulting in the K-&gt;N amino acid substitution (exemplified by sequence HUMNKSFP40 (Wolf et al., 1991)) could not be found in DNA representing 224 chromosomes, including 97 which had the same IL12p40 allele as HUMNKSFP40, as defined by the presence of the 3UTR TaqI (not shown). Similarly, neither the exon 7 difference nor the promoter differences between the PAC and the published sequences could be confirmed.  
         [0135]    Further Search for polymorphisms in the IL-12 p40 Gene  
         [0136]    DNA from different individuals representative of the TaqI− and TaqI+ alleles was tested for further differences in and around the IL-12p40 gene. Polymorphisms were sought by PCR amplification followed by SSCP, restriction enzyme digestion or direct sequencing. Variants found by SSCP were confirmed by subsequent sequencing (or other methods as appropriate) of samples from a number of unrelated individuals. The results are summarized in Tables 9 and 10. Our major interest was in finding whether commonly occurring IL-12p40 polymorphisms exist as these may be useful for testing in various disease situations. It should be noted that the possibility of other, rarer, IL12p40 variants was not tested and is not excluded.  
         [0137]    Despite extensive searching, no coding region sequence differences were found. Simple sequence repeat polymorphisms were discovered in introns 2 and 4. However, these had limited heterogeneity, with only two and three alleles found for each, respectively. A number of apparent single nucleotide substitutions were found. All the polymorphisms listed in Table 9 and 10 were confirmed by sequencing. Some polymorphisms were examined further in a large sample of unrelated individuals of diverse European descent. In particular, the 3′UTR alleles were found to be in Hardy-Weinberg equilibrium, with the TaqI− allele having a frequency of 0.82. The TA repeat polymorphism in intron 4 showed a similar distribution, also in Hardy-Weinberg equilibrium. The longest allele of this polymorphism is probably in linkage disequilibrium with the 3′UTR TaqI+allele. Even though the sample size was small, there was a suggestion that other polymorphisms may not be in linkage disequilibrium, notably the two single base changes, each A-&gt;G, within 14 bp in intron 2 (Table 9, 10). In sequencing this region from 8 individuals, 2 were heterozygous for either form, while of the homozygotes, 5 had the AA haplotype and 1 each the AG and GG haplotypes.  
         [0138]    There was clustering of the DNA sequence variations. Of the twelve polymorphisms found, four were in intron 1, three were in intron 2, and two in intron 4. The changes in introns 1 and 2 appeared in pairs separated by no more than 60 bp. No polymorphisms were found (by SSCP analysis) in introns 5 and 6. In searching for genetic variants in 1L-12p40, no changes were found which would give rise to amino acid changes. An apparent amino acid substitution in comparison with a previously described cDNA sequence could not be found in testing an additional 128 chromosomes. The dearth of any coding sequence changes indicates a high level of conservation between the human subjects tested. Perhaps it was not surprising that no sequence variants were found that resulted in amino acid substitutions, given that IL-12p40 plays a fundamental role in immune regulation (Trinchieri G., 1995). However, this contrasts with the large number of differences displayed between species: there are 116 differences in amino acid sequences between the approximately 335 residues of the mouse and human IL-12p40 proteins (Gubler et al., 1991; Wolf et al., 1991; Tone et al., 1996). Despite the sequence differences, the genomic organization of mouse (Wolf et al., 1991) and human IL-12p40 genes is similar: both have 8 exons and the relative size of the introns is similar in both species. The mouse gene has an untranslated first exon but, unlike the human gene, the last exon does encode part of the final protein product (Wolf et al., 1991).  
         [0139]    The polymorphisms described are useful in genetic studies to determine the role of IL-12p40 in regulation of the immune response in health and disease.  
         [0140]    Method  
         [0141]    Polymorphisms in the IL12p40 gene were sought. DNA fragments covering the entire gene were amplified from a panel of up to 27 unrelated donors and S was performed as follows. Forward and reverse primers shown were selected from the PAC sequence, and used to amplify specific segments of the IL-1 gene. “Standard” PCR conditions were performed incorporating 32 P-dATP: 2′ at 95° C. followed by 35 cycles of 20 s at 95° C., 20 s at 55° C., and 30 s at different extension times are indicated. For SSCP, a portion (1 ml) of the reaction mix was added to 5 ml of loading buffer (95% formamide, 20 mM EDTA, NaOH, 0.05% bromophenol blue and 0.15% xylene cyanol) heated at 90° C. for 1 min. and loaded onto a 4 to 5% polyacrylamide gel (1:45 or 1:90 ratio of N-methylene bisacrylamide to acrylamide). The intron 7 product was digested with EcoRV and HindIII prior to SSCP. Electrophoresis was performed overnight at room temperature. The gel was blotted on filter paper and exposed to autoradiography overnight at −70° C. Fragments which gave variable products were selected for sequencing. Note: the sequencing panel was selected so as to be enriched for individuals homozygous for the exon 8 TaqI allele number of individuals sequenced who were homozygous for the canonical PAC sequence, or for the alternate non-PAC sequence are shown, as is the number heterozygotes.  
       EXAMPLE 7  
     Linkage Disequilibrium of Regulatory IL12P40 Alleles with Type I Diabetes  
       [0142]    Results  
         [0143]    To test whether IL12B may be a susceptibility gene in human T1D, 249 sibpairs were typed for markers on chromosome 5q33-34, to which IL12B was mapped (Warrington et al., 1994). Testing multiplex families for markers from this region initially resulted in a modest lod score, suggestive of linkage to a susceptibility gene (FIG. 3). Stratification of sibpairs has proven useful in revealing linkage in multipoint analyses, allowing clear definition of the susceptibility locus IDDM13 (Morahan et al., 1996; Fu et al., 1998; Larsen et al., 1999). Applying stratification to the 5q data revealed a difference between sibpairs sharing HLA haplotypes and those differing at HLA (FIG. 3). The HLA-identical sibpairs showed linkage to this region with a maximized lod greater than 2.3; this susceptibility locus is provisionally amed IDDM18. In contrast, and unlike the case for IDDM13 (Morahan et al., 1996), there was no evidence of linkage in the HLA mismatched sibs. This emphasizes the genetic heterogeneity of TID, such that different subgroups have susceptibility arising from different interactions of HLA and non-HLA genes.  
         [0144]    The linkage analyses indicated that IDDM18 may reside near IL12B. The complete sequence of, and genetic polymorphisms in and around, IL12B have been described (Huang et al., in press). Although there were no common coding region variants, we found useful polymorphisms in the 3′ UTR, intron 4 and the promoter. These polymorphisms were typed and the transmission disequilibrium test (Spielman et al., 1993) TDT was applied. There was significant excess transmission of particular intron 4 and 3′ UTR alleles, but not of alleles defined by the promoter polymorphism (Table 13). (The intron 4 and 3′ UTR alleles are in linkage disequilibrium, so further discussion is limited to the latter). A physical map of &gt;1 Mb surrounding IL12B was constructed (FIG. 4 a ) and searched for futher downstream polymorphisms; one resulting marker, D5S2937, has 10 alleles, none of which singly or jointly generated significant TDT results (Table 13). These observations were confirmed using the T sp  statistic, which adjusts for testing more than one affected subject per family (Martin et al., 1998) (Table 13). Similar results were also obtained testing only one affected sib per family (data not shown).  
         [0145]    To further test the involvement of IL12B polymorphisms in T1D, the families were divided into two groups: one showing linkage to 5q33-34 (which would be expected to shown the influence of the disease allele) and one which did not (and would therefore predominantly include sibs who had TID due to other susceptibility loci). The TDT was applied to each group (FIG. 4 b ). Evidence for preferential transmission increased in the linked group, whereas there was no significant deviation in transmission of alleles to the unlinked group. (Although this method of selection of sibs for linkage will affect matching of alleles within families, it should not affect genotypes between families, and hence should not affect the overall TDT. In fact, similar results were obtained when the analysis was restricted to the first affected sib (data not shown). The results showed preferential transmission in only those families in which T1D was linked to IL12B, indicating that TID is mediated in part by the IL-12-linked causative polymorphism. There was again preferential transmission of the 3′ UTR polymorphism, but none at the promoter polymorphism only 20 kb upstream (FIGS. 4 a,b ).  
         [0146]    If the 3′ UTR polymorphism itself contributes to susceptibility, the offspring of homozygous parents should not show linkage, unlike offspring of heterozygous parents (Robinson et al., 1993). Because the frequency of the susceptibility allele is 0.8, the families in which at least one parent is homozygous will be in the majority, helping to explain the low lod scores obtained in the original analysis. Essentially, all the evidence for linkage (MLS=2.632) was maintained in the group with at least one heterozygous parent; there was no evidence for linkage to IL12B promoter alleles in families in which both parents were homozygous at the 3′ UTR (MLS=0.388).  
         [0147]    It was crucial to confirm the above findings of preferential transmission of IL12B 3′ UTR allele 1 to T1D subjects. The Australian IDDM DNA Repository has been established and into which 238 families have been recruited and typed for IL12B-associated polymorphisms. The results confirm those obtained above: preferential transmission of allele 1 of the 3′ UTR polymorphism, and lack of bias in transmission of promoter alleles (Table 14). The Australian IDDM DNA Repository families were also typed at a novel polymorphism, DS2340, located 12 kb downstream of the IL12B 3′ UTR. This marker did not yield significant TDT results (Table 14). Combining the results from both TDT analyses of the IL12B 3′ UTR, the null hypothesis of lack of association of the IL12B 3′ UTR with TID may be rejected (overall P=3.5×10 7 ). Significant linkage disequilibrium appears confined to a region of approximately 30 kb in which IL12B is the only known gene.  
         [0148]    The results show that the 3′ UTR allele 1 is preferentially transmitted to T1D subjects, and hence either itself confers susceptibility or is in linkage disequilibrium with the disease-predisposing variant; allele 2 is preferentially non-transmitted, so it may be associated with T1D resistance. As no common change was found in its coding sequences, if IL12B is involved in TID susceptibility then its alleles should show some other functional difference. To address this, EBV-transformed cell lines (which are known to express IL-12; refs. Wolf et al., 1991; Gubler et al., 1991) homozygous for each allele were identified. Expression of IL12B was significantly reduced in the 2/2 genotype cell line relative to the 1/1 line (FIG. 5). The 3′ UTR polymorphism is located over 1 kb from the mRNA degradation element (Zubiaga et al., 1995), so it is unlikely that the observed difference between the cell lines is due to differences in stability. The inference that the 3′ UTR polymorphism may affect gene expression is supported by a similar finding for the rat gene spi2.3 (LeCam et al., 1995). If differences in IL12B expression result in different levels of protein, then individuals with the susceptibility allele should produce more IL12p40. Higher IL-12 levels were found in relatives of T1D probands (Szelachowska et al., 1997). Increased IL-12 may promote Th1 cells, and aggravate autoimmune destruction of β-cells, causing T1D (as in NOD mice (Katz et al., 1995; Trembleau et al., 1995)). In contrast, lower levels of IL-12 should reduce susceptibility because IL-12 antagonists can protect NOD mice from diabetes (Trembleau et al., 1997).  
         [0149]    Materials and Methods  
         [0150]    Genotyping  
         [0151]    We tested a total of 249 affected sibpairs, including families that were previously described (Morahan et al., 1996) and an additional 120 families obtained from the British Diabetes Association. An additional independent cohort of 235 predominantly simplex families was also recruited into the Australian IDDM DNA Repository. DNA from individuals from multiplex families were typed using either anonymous microsatellite markers (Weissenbach et al., 1992) or the highly polymorphic repeat within GABRA1 (Johnson et al., 1992). We tested polymorphisms in and around IL12B as described (Huang et al., in press). The D5S2937 marker is a simple sequence repeat which was generated from inspection of the draft sequence of the BAC 9p16 from 5q33-34 obtained from the DOE&#39;s Joint Genome Institute (ftp://ft]2.1gi-psf.orgipub/JGI-data/Human/Ch5/Draft/). primers to amplify this TAA repeat were 5′-GGGTAAGCGATTCAAA-CATT-3′ (&lt;400&gt;137) (forward) and 5′GGTATTGCATTGTAGGCACAT-3′ (&lt;400&gt;138) (reverse). D5S2940 is a C(T)n repeat located 12 kb centromeric of the 3′ UTR and was amplified with primers 5′GGGCAACAAGAGTGAAACT-3′ (&lt;400&gt;139) and 5′-TCAAAAGAGGTCCGTCTAAA-3′ (&lt;400&gt;140).  
         [0152]    Genetic Analyses  
         [0153]    We carried out multipoint linkage analysis using the MAPMAKER/Sibs software program (Kruglyak et al., 1995), and TDT analyses (Spielman et al., 1993) using both the GAS software package (Young, A., 1994) and Tsp program (Martin et al., 1998).  
         [0154]    Gene Expression  
         [0155]    We typed EBV-transformed cell lines from the 4th Asia-Oceania Histocompatibility Workshop cell line panel Degli-Esposti et al., 1993) for the IL12B 3′ UTR allele, and cell lines representing the 1/1 and 2/2 genotypes were selected. We isolated total RNA from these cell lines using guanadinium thiocyanate and purified it by CsCl-density gradient centrifugation. Northern-blot analysis was performed by standard methods (Sambrook et al., 1989) with human IL12B and GAPDH cDNA probes. The levels of IL12B MRNA in each cell line relative to GAPDH was determined by densitometry in three separate experiments. Similar results were obtained by RT-PCR (data not shown).  
       EXAMPLE 8  
     Polymorphisms Haplotypes as a Marker of Disease Susceptibility or Resistance  
       [0156]    The combination of particular polymorphisms is used to define IL12B haplotypes. These haplotypes are used to test for susceptibility or resistance to immune related diseases in which IL12 production and/or Th1-Th2 regulation may be relevant.  
         [0157]    An example of the use of such haplotypes is demonstrated in the table below. Combining promoter and 3′ UTR alleles generates 4 haplotypes. The appearance of these haplotypes may be compared between different groups which differ in a relevant phenotype. To illustrate this point, consider subjects with diabetes and first degree relatives who do not have diabetes but who have autoantibodies (i.e. “preclinical”). The table shows that there is a difference in the proportion of haplotype C homozygous individuals in these groups. Thus haplotypes may be used to predict likelihood to proceed from early autoimmunity to diabetes. Haplotypes may be used in this way to test for susceptibility or resistance to other disease conditions or predisposition to mounting Th1 or Th2 type immune responses.  
         [0158]    Haplotype analysis of IDDM and preclinical subjects.  
                                                                                 Subjects   Haplotype C/C   Other haplotypes   P                                        IDDM   122   124   0.005           Preclinical   9   32                      
 
         [0159]    Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.  
                                               TABLE 7                           Sequences of Exon-Intron boundaries.            Intron   Sequence                        1 &lt;400&gt;116     GCCCAGAGCAAG GTAAGCACTTCC . . . CCCTTCTTATAG ATG TGTCACCAG     &lt;400&gt;117                   2 &lt;400&gt;118     TGAAGAAAGATG GTAACCAGCCTC . . . TGTGCATTCCAG TTTATGTCGTAG     &lt;400&gt;119               3 &lt;400&gt;120     AGGACCAGAAA GGTAATTCTATAC . . . TTTCAAATCCAG AACCCAAAAATA     &lt;400&gt;121               4 &lt;400&gt;122     AAGCAGCAGAGG GTGAGTGAAACT . . . CTTTGACTTCAG CTCTTCTGACCC     &lt;400&gt;123               5 &lt;400&gt;124     TCAGGGACATCA GTGAGTTTTGGA . . . CCTCTTCCACAG TCAAACCTGACC     &lt;400&gt;125               6 &lt;400&gt;126     AAGAGAGAAAAG GTAAGAAGTGAT . . . TCTCTTTTGCAG AAAGATAGAGTC     &lt;400&gt;127               7 &lt;400&gt;128     CCCTGCAGT TAG GTGAGCAGGCCC . . . ATTCTCTTCCAG GTTCTGATCCAG     &lt;400&gt;129                  
 
         [0160]    Sequences of Exon-Intron boundaries. The complete IL12p40 genomic sequence was determined using PAC 93.1 as a template. The sequence has been deposited in Genbank. Exon-intron boundaries were determined by comparison with published CDNA sequences (Gubler et al., 1991; Wolf et al., 1991). Exon sequences are shown in bold; splice donor and acceptor sites are single underlined. The start and stop codons (double underlined) are the first and last codons in exons 2 and 7 respectively. Sizes of introns are indicated in FIG. 1C.  
                                                                       TABLE 8                           Comparison of genomic and published IL12p40 sequences            Region   DNA source   Position       Sequence   Comment                    Promoter   PAC 93-1   544   &lt;400&gt;3   AGTTTTTTTTTTTTAATTTTCAAGGTGCTT                                       ----------------*-------------   No RE difference                   HSU89323       &lt;400&gt;4   AGTTTTTTTTTTTTAAATTTCAAGGTGCTT   Could not confirm               Promoter   PAC 93-1   885   &lt;400&gt;5   AACATACCTGCAATCTGCTTTGTCCACTTA                               -----------------*------------   No RE difference                   HSU89323       &lt;400&gt;6   AACATACCTGCAATCTGATTTGTCCACTTA   Could not confirm               Promoter   PAC 93-1   1973   &lt;400&gt;7   CTAAACCCTTTGCCCTTCATCTCATCCTC                               --------------*--------------   No RE difference                   HSU89323       &lt;400&gt;8   CTAAACCCTTTGCC-TTCATCTCATCCTC   Could not confirm               Exon 6   PAC 93-1   14041   &lt;400&gt;9   TCAAACCTGACCCACCCAAGAACTTGCAGC   HUMCLMF40 = PAC                               -------------------*----------   Gives K -&gt; N change                   HUMNKSFP40       &lt;400&gt;10   TCAAACCTGACCCACCCAACAACTTGCAGC   Could not confirm               Exon 7   PAC 93-1   16117   &lt;400&gt;11   AGATAGAGTCTTCACGGACAAGACCTCAGC   HUMCLMF40 = PAC                               ---------------*--------------   Silent change                   HUMNKSFP40       &lt;400&gt;12   AGATAGAGTCTTCACCGACAAGACCTCAGC   Could not confirm               3′ UTR   PAC 93-1   16974   &lt;400&gt;13   TTGTATAGTTAGATGCTAAATGCT   L3 same; TaqI−               (exon 8)               ----------*-------------   Creates a TaqI site                   HUMNKSFP40       &lt;400&gt;14   TTGTATAGTTCGATGCTAAATGCT   L26 same; TaqI+                                  
 
         [0161]    [0161]                                                                                                                                                           TABLE 9                       Detection and Confirmation of IL12p40 polymorphisms                                    Product   Reaction                Polymorphism   Primers (5′-&gt;3′)   size   conditions*                    Intron 1 3696   GGCTTAAAGGGGCCAAGT &lt;400&gt;15   401   Standard                       AGGGAGCACTATCCCTCAGC &lt;400&gt;16               Intron 1 3757   GGCTAAAGGGGCCAAGT &lt;400&gt;17   401   Standard                   AGGGAGCACTATCCCTCAGC &lt;400&gt;18               Intron 1 4572   ATGTTATCTCATTGCCTTC &lt;400&gt;19   511-513   Standard                   AAGTGGTTCTGAAACCACTG &lt;400&gt;20               Intron 1 4793   ATGTTATCTCATTGCCTTTC &lt;400&gt;21   511-513   Standard                   AAGTGGTTCTGAAACCACTG &lt;400&gt;22               Intron 2 8798   GGGAAGACTAAGCTCTACTG &lt;400&gt;23   128-131   Standard                   GGATTTCGTTCCCTCTGTTT &lt;400&gt;24               Intron 2 8930   GGGAAGACTAAGCTCTACTG &lt;400&gt;25   413   Standard                   CAACGAACCAAGACTGTCAT &lt;400&gt;26               Intron 2 8944   GGGAAGACTAAGCTCTACTG &lt;400&gt;27   413   Standard                   CAACGAACCAAGACTGTCAT &lt;400&gt;28               Intron 3 9910   TTCTAAGCCATTCGCTCCTG &lt;400&gt;29   188   Standard                   GTTAATTCATTACACTCACC &lt;400&gt;30               Intron 4 11244   TACTTCTGCTGACACCACTA &lt;400&gt;31   436   Standard                   GAACTAGGATCAAATTGTATAC &lt;400&gt;32               Intron 4 11563   GGTTACATAATCATATGTA &lt;400&gt;33   254-258   Standard                   GTTAGGATTTCAGGTGTGAG &lt;400&gt;34               Intron 7 16521   TAGCTCATCTTGGAGCGAAT &lt;400&gt;35   982   70 s at 72° C.                   AACATTCCATACATCCTGGC &lt;400&gt;36               Exon 7 16117   GAAAGGCCATGCACCTAAC &lt;400&gt;37   1,285   90 s at 72° C.                   TCCAGGTGCACTGAGAGT &lt;400&gt;38               Exon 8 16974   TTTGGAGGAAAAGTGGAAGA &lt;400&gt;39   300   2′ final at 72° C.                   AACATTCCATACATCCTGGC &lt;400&gt;40       TaqI digestion                    Detection   Number   Number   Number homozygous for   Number                Method   screened   sequenced   PAC allele   non-PAC allele   Heterozygous                    SSCP   23   16   1   11   4                   SSCP   23   16   3   10   3               SSCP   27   15   1   12   2               SSCP   27   13   2   10   1               SSCP   27   6   4   2   0               SSCP   27   9   1   7   1               SSCP   27   8   2   5   1               SSCP   27   9   3   5   1               SSCP   24   14   2   12   0               SSCP   see Table 5               EcoRV, HindIII       15   1   14   0               digest, SSCP               direct sequence       9   9   0   0                   see Table 5                    
         [0162]    [0162]                                                               TABLE 10                           Allelic differences in IL12p40 sequences.            Intron   DNA   Position   Sequence   No. Alleles                    1   L26       TCTTTAATAATAACTCCCTTTT   &lt;400&gt;41                               ---------*------------                   PAC   3696   TCTTTAATAGTAACTCCCTTTT   &lt;400&gt;42                   L26       GCCCACCCAAGTGTCATTGG   &lt;400&gt;43                           -----------*--------                   PAC   3757   GCCCACCCAAGCGTCATTGG   &lt;400&gt;44                   L26       TCAAGCCTG-TCTGTTTAA   &lt;400&gt;45                           ---------**---------                   PAC   4572-3   TCAAGCCTGTGTCTGTTTAA   &lt;400&gt;46                   L26       CCGCCTAGACTTAGTAG   &lt;400&gt;47                           ---------*-------                   PAC   4793   CCGCCTAGAGTTAGTAG   &lt;400&gt;48               2   L26       TAAAAATAATAATAATAATAATAATAATAATG   &lt;400&gt;49 2                           ------***-----------------------   2                   PAC   8798-8800   TAAAAA---TAATAATAATAATAATAATAATG   &lt;400&gt;50                   L26       CTCCTCAGTCTATAAGTAACAATAACTA   &lt;400&gt;51                           ------*-------------*-------                   PAC   8930; 8944   CTCCTCGGTCTATAAGTAACGATAACTA   &lt;400&gt;52               3   L26       CGCTCATAAGGGTTAAAAACAACAACAAC   &lt;400&gt;53                           ----------*------------------                   PAC   9910   CGCTCATAAGAGTTAAAAACAACAACAAC   &lt;400&gt;54               4   L26       TCTCCAAGTGCAAAAAGACATAATCAGCAG   &lt;400&gt;55                           ------------*-----------------                   PAC   11244   TCTCCAAGTGCATAAAGACATAATCAGCAG   &lt;400&gt;56                   L26       TATATATATAT----AAAATGTGTATACA   &lt;400&gt;57 3                           -----------****--------------                   PAC   11563-6   TATATATATATATATAAAATGTGTATACA   &lt;400&gt;58               7   L26       AGAGCATGGAGGACTTGCA   &lt;400&gt;59                           ---------*---------                   PAC   16521   AGAGCATGGCGGACTTGCA   &lt;400&gt;60                            
         [0163]    [0163]                                                                                 TABLE 11                           Allele Frequencies of IL12p40 variants.                    Hardy-           Frequency   Weinberg            Polymorphism   No. tested   Allele 1   Allele 2   Allele 3   equilibrium                    Intron 4 11563   336   0.79   0.2   0.01   Yes       Exon 8 16974   382   0.82   0.18       Yes                    
         [0164]    Allele frequencies were determined by genotyping unrelated subjects of diverse European descent. The intron 4 TA repeat polymorphism was detected on denaturing bis-acrylamide gels. The exon 8 TaqI allele was detected as follows: 2 ul of products digested with 1 unit of TaqI in a 1o ul reaction volume and incubate at 65° C. for 2 hours. The number of individuals with each genotype did not differ from that expected if the alleles were in Hardy-Weinberg equilibrium.  
                                                           TABLE 12                           Primers used for sequencing PAC93-1                Forward primers:   Reverse primers:                        &lt;400&gt;61   PF   GGAAGGCGCCCCAGATGTA   P9R   GAATTGCATGCTGGGTTCTG   &lt;400&gt;86                   &lt;400&gt;62   P3F   TCAGACACATTAACCTTGCA   P8R   CTGTGGCTTCCAGAGGTTAC   &lt;400&gt;87               &lt;400&gt;63   P4F   CAATAGACAAGTGATTTCACTG   P7R   TAATGTGGTCATTGGCAGGT   &lt;400&gt;88               &lt;400&gt;64   P6F   ATGCTTAATTATAACTATATTC   P5R   CAGATGAGTCCTTGTGCCCC   &lt;400&gt;89               &lt;400&gt;65   P7F   CCATAATAGGTTCATTGCCC   P4R   ACCCGGGCCAGAGCAGCG   &lt;400&gt;90               &lt;400&gt;66   Int1-3F   GGCTTAAAGGGGCCAAGT   P3R   GCGGAATAAAGATATCTCTC   &lt;400&gt;91               &lt;400&gt;67   Int1-6F   CCCACCACCATCACCTCT   P2R   GATGAGATGAAGGCAAAGG   &lt;400&gt;92               &lt;400&gt;68   Int1-5F   ATGTTATCTCATTGCCTTTC   PR   TCACCAGGGATGCTTCCAGG   &lt;400&gt;93               &lt;400&gt;69   Int1 (F)   GAAGAAAGGGGAGAATCAAG   Int1-5R   GGTTACCCACATTCCATC   &lt;400&gt;94               &lt;400&gt;70   Int2F   GGAAAATGCAATGCCATATC   Int1-2Fr   CTTATGCCATGGATCATGTC   &lt;400&gt;95               &lt;400&gt;71   Int2-1F   ATCCTGAATTTCCTCAACTG   Int1-7R   AAGTGGTTCTGAAACCACTG   &lt;400&gt;96               &lt;400&gt;72   Int2-2F   AGAGACTGTCTGTATCCCAT   Int1-4R   CCAGAGTGTCTGATTCAGC   &lt;400&gt;97               &lt;400&gt;73   Int2-4F   AGGCCTGAGCCAGGGGTAT   Int1-3R   AAGGCAAGCCATCTGATACA   &lt;400&gt;98               &lt;400&gt;74   Ex3F   TTCTAAGCCATTCGCTCCTG   Int2R   AGGCCAACGATCTAAGCATG   &lt;400&gt;99               &lt;400&gt;75   Int4F   ATTCTGGACGTTTCACCTGC   Int2-1R   GGAGTGGCAGAGGCCTGG   &lt;400&gt;100               &lt;400&gt;76   Int4F-4   TACTTCTGCTGACACCACTA   Ex3-R   CACCATTTCTCCAGGGGCA   &lt;400&gt;101               &lt;400&gt;77   F-Int4.3   GGATGAAGGAGACATACACT   Ex3-1R   CAACGAACCAAGACTGTCAT   &lt;400&gt;102               &lt;400&gt;78   IL-12A   GCCGTTCACAAGCTCAAGTA   Int3-1R   GTTAATTCATTACACTCACC   &lt;400&gt;103               &lt;400&gt;79   Int5F   GGACTTCTTTCTTAGAATAT   Int4-8R   ATAGGTCACTGAGAGGTTGC   &lt;400&gt;104               &lt;400&gt;80   IL-12E   ATCAAACCTGACCCACCCAA   Int4-3R   AGCTTGTTGTATCCTTCCAG   &lt;400&gt;105               &lt;400&gt;81   Int6F   ATGTGATCCTTCTTTGACTG   Int4R   TCATACTCCTTGTTGTCCCC   &lt;400&gt;106               &lt;400&gt;82   Int6-1F   GAAAGGCCATGCACCTAAC   5-1R   CGGCTAGCTGTAAGATCTGA   &lt;400&gt;107               &lt;400&gt;83   IL-12TAQF   TAGCTCATCTTGGAGCGAAT   Int5R   CATGGAACTAAGCTGAGCCC   &lt;400&gt;108               &lt;400&gt;84   #6   TTTGGAGGAAAAGTGGAAGA   IL-12 * R   CGCAGAATGTCAGGGAGAAG   &lt;400&gt;109               &lt;400&gt;85   T7   AATACGACTCACTATAG   Int6R   TCCAGGTGCACTGAGAGT   &lt;400&gt;110                           Int6-1R   TTCTAGCACAATTGCCTTGCCT   &lt;400&gt;111                           IL-12B   AACATTCCATACATCCTGGC   &lt;400&gt;112                           Ex8-1R   GCAGGAAGACACTGACTTTG   &lt;400&gt;113                           Ex8-2R   GCCTTCCAGACACTTACGGT   &lt;400&gt;114                           T3   ATTAACCCTCACTAAAG   &lt;400&gt;115                          
 
         [0165]    [0165]                                                                                                 TABLE 13                       Analyses of allelic transmission to affected offspring                   TDT of polymorphisms in IL12B            Polymorphism   Allele   Freq.   Trans   Not   P               3′ UTR   1   0.79   171   122   0.0025           2   0.21   122   171   —       intron 4   1   0.79   137    85   0.00029           2   0.20    85   131   —           3   0.01    0    6   —       promoter   1   0.56   176   191   0.77           2   0.44   191   176   —                    TDT at a centromeric locus, D5S2937            Allele   Freq   Trans.   Not   P               1   0.02    4    6   —       2   0.07   30   14   0.011       3   0.10   30   50   —       4   0.23   67   76   —       5   0.07   19   14   —       6   0.17   63   61   —       7   0.04   17   10   —       8   0.21   74   66   —       9   0.09   24   31   —                    TDT allowing for multiple affected family members (T sp )                Marker   DF   χ 2     P                       D5S2937   8   18.292   0.0191           3′ UTR   1   12.694   0.0004           Intron 4   2   10.549   0.0051           promoter   1   0.305   0.5809                        
         [0166]    TDT analyses (Spielman et al., 1993) of polymorphisms in and around IL12B were carried out on the data from the linkage study (FIG. 3). Markers are shown in order from centromere to telomere (Huang et al., in press). The D5S2937 marker is a simple sequence repeat which was generated from inspection of the draft sequence of a cosmid from 5q33-34. This marker was placed on the physical map in relation to IL12B. Note that no correction was made for testing multiple alleles at this locus. For clarity , only P&lt;0.1 is shown. The data were used to calculate the T sp  statistics shown, which corrects for multiple affected individuals per family (Martin et al., 1998). DF, degrees of freedom. No correction was made for testing multiple alleles at the D5S2937 locus.  
                                                                   TABLE 14                           TDT of an independent cohort of simplex families            Polymorphism   Allele   Freq   Trans   Not Trans.   P                    IL12B promoter   1   0.5   101   96   —           2   0.5   96   101   —       IL12B 3′ UTR   1   0.78   101   55   0.00014           2   0.22   55   101   —       D5S2940   1   0.04   7   12   —           2   0.5   93   97   —           3   0.45   98   89   —           4   0.01   5   5   —                  
 
         [0167]    235 simplex families with one affected child were genotyped for the IL12B promoter or 3′ UTR alleles, as well as for D5S2940. Note that although 235 families were tested, the high homozygosity rate for the 3′ UTR polymorphism meant that most parents were not informative for this marker. Allele frequencies were calculated based on parental genotypes. For clarity, only P&lt;0.1 is shown.  
       BIBLIOGRAPHY  
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         1 
         
           
             140  
           
           
             1  
             419  
             DNA  
             mammalian  
           
            1 

cagcattagc gtgcgggccc aggaccgcta ctatagctca tcttggagcg aatgggcatc     60 

tgtgccctgc agttaggttc tgatccagga tgaaaatttg gaggaaaagt ggaagatatt    120 

aagcaaaatg tttaaagaca caacggaata gacccaaaaa gataatttct atctgatttg    180 

ctttaaaacg tttttttagg atcacaatga tatctttgct gtatttgtat agttcgatgc    240 

taaatgctca ttgaaacaat cagctaattt atgtatagat tttccagctc tcaagttgcc    300 

atgggccttc atgctattta aatatttaag taatttatgt atttattagt atattactgt    360 

tatttaacgt ttgtctgcca ggatgtatgg aatgtttcat actcttatga cctgatcca     419 

 
           
             2  
             419  
             DNA  
             mammalian  
           
            2 

cagcattagc gtgcgggccc aggaccgcta ctatagctca tcttggagcg aatgggcatc     60 

tgtgccctgc agttaggttc tgatccagga tgaaaatttg gaggaaaagt ggaagatatt    120 

aagcaaaatg tttaaagaca caacggaata gacccaaaaa gataatttct atctgatttg    180 

ctttaaaacg tttttttagg atcacaatga tatctttgct gtatttgtat agttagatgc    240 

taaatgctca ttgaaacaat cagctaattt atgtatagat tttccagctc tcaagttgcc    300 

atgggccttc atgctattta aatatttaag taatttatgt atttattagt atattactgt    360 

tatttaacgt ttgtctgcca ggatgtatgg aatgtttcat actcttatga cctgatcca     419 

 
           
             3  
             30  
             DNA  
             mammalian  
           
            3 

agtttttttt ttttaatttt caaggtgctt                                      30 

 
           
             4  
             30  
             DNA  
             mammalian  
           
            4 

agtttttttt ttttaaattt caaggtgctt                                      30 

 
           
             5  
             30  
             DNA  
             mammalian  
           
            5 

aacatacctg caatctgctt tgtccactta                                      30 

 
           
             6  
             30  
             DNA  
             mammalian  
           
            6 

aacatacctg caatctgatt tgtccactta                                      30 

 
           
             7  
             29  
             DNA  
             mammalian  
           
            7 

ctaaaccctt tgcccttcat ctcatcctc                                       29 

 
           
             8  
             28  
             DNA  
             mammalian  
           
            8 

ctaaaccctt tgccttcatc tcatcctc                                        28 

 
           
             9  
             30  
             DNA  
             mammalian  
           
            9 

tcaaacctga cccacccaag aacttgcagc                                      30 

 
           
             10  
             30  
             DNA  
             mammalian  
           
            10 

tcaaacctga cccacccaac aacttgcagc                                      30 

 
           
             11  
             30  
             DNA  
             mammalian  
           
            11 

agatagagtc ttcacggaca agacctcagc                                      30 

 
           
             12  
             30  
             DNA  
             mammalian  
           
            12 

agatagagtc ttcaccgaca agacctcagc                                      30 

 
           
             13  
             24  
             DNA  
             mammalian  
           
            13 

ttgtatagtt agatgctaaa tgct                                            24 

 
           
             14  
             24  
             DNA  
             mammalian  
           
            14 

ttgtatagtt cgatgctaaa tgct                                            24 

 
           
             15  
             18  
             DNA  
             mammalian  
           
            15 

ggcttaaagg ggccaagt                                                   18 

 
           
             16  
             20  
             DNA  
             mammalian  
           
            16 

agggagcact atccctcagc                                                 20 

 
           
             17  
             18  
             DNA  
             mammalian  
           
            17 

ggcttaaagg ggccaagt                                                   18 

 
           
             18  
             20  
             DNA  
             mammalian  
           
            18 

agggagcact atccctcagc                                                 20 

 
           
             19  
             20  
             DNA  
             mammalian  
           
            19 

atgttatctc attgcctttc                                                 20 

 
           
             20  
             20  
             DNA  
             mammalian  
           
            20 

aagtggttct gaaaccactg                                                 20 

 
           
             21  
             20  
             DNA  
             mammalian  
           
            21 

atgttatctc attgcctttc                                                 20 

 
           
             22  
             20  
             DNA  
             mammalian  
           
            22 

aagtggttct gaaaccactg                                                 20 

 
           
             23  
             20  
             DNA  
             mammalian  
           
            23 

gggaagacta agctctactg                                                 20 

 
           
             24  
             20  
             DNA  
             mammalian  
           
            24 

ggatttcgtt ccctctgttt                                                 20 

 
           
             25  
             20  
             DNA  
             mammalian  
           
            25 

gggaagacta agctctactg                                                 20 

 
           
             26  
             20  
             DNA  
             mammalian  
           
            26 

caacgaacca agactgtcat                                                 20 

 
           
             27  
             20  
             DNA  
             mammalian  
           
            27 

gggaagacta agctctactg                                                 20 

 
           
             28  
             20  
             DNA  
             mammalian  
           
            28 

caacgaacca agactgtcat                                                 20 

 
           
             29  
             20  
             DNA  
             mammalian  
           
            29 

ttctaagcca ttcgctcctg                                                 20 

 
           
             30  
             20  
             DNA  
             mammalian  
           
            30 

gttaattcat tacactcacc                                                 20 

 
           
             31  
             20  
             DNA  
             mammalian  
           
            31 

tacttctgct gacaccacta                                                 20 

 
           
             32  
             22  
             DNA  
             mammalian  
           
            32 

gaactaggat caaattgtat ac                                              22 

 
           
             33  
             19  
             DNA  
             mammalian  
           
            33 

ggttacataa tcatatgta                                                  19 

 
           
             34  
             20  
             DNA  
             mammalian  
           
            34 

gttaggattt caggtgtgag                                                 20 

 
           
             35  
             20  
             DNA  
             mammalian  
           
            35 

tagctcatct tggagcgaat                                                 20 

 
           
             36  
             20  
             DNA  
             mammalian  
           
            36 

aacattccat acatcctggc                                                 20 

 
           
             37  
             19  
             DNA  
             mammalian  
           
            37 

gaaaggccat gcacctaac                                                  19 

 
           
             38  
             18  
             DNA  
             mammalian  
           
            38 

tccaggtgca ctgagagt                                                   18 

 
           
             39  
             20  
             DNA  
             mammalian  
           
            39 

tttggaggaa aagtggaaga                                                 20 

 
           
             40  
             20  
             DNA  
             mammalian  
           
            40 

aacattccat acatcctggc                                                 20 

 
           
             41  
             22  
             DNA  
             mammalian  
           
            41 

tctttaataa taactccctt tt                                              22 

 
           
             42  
             22  
             DNA  
             mammalian  
           
            42 

tctttaatag taactccctt tt                                              22 

 
           
             43  
             20  
             DNA  
             mammalian  
           
            43 

gcccacccaa gtgtcattgg                                                 20 

 
           
             44  
             20  
             DNA  
             mammalian  
           
            44 

gcccacccaa gcgtcattgg                                                 20 

 
           
             45  
             18  
             DNA  
             mammalian  
           
            45 

tcaagcctgt ctgtttaa                                                   18 

 
           
             46  
             20  
             DNA  
             mammalian  
           
            46 

tcaagcctgt gtctgtttaa                                                 20 

 
           
             47  
             17  
             DNA  
             mammalian  
           
            47 

ccgcctagac ttagtag                                                    17 

 
           
             48  
             17  
             DNA  
             mammalian  
           
            48 

ccgcctagag ttagtag                                                    17 

 
           
             49  
             32  
             DNA  
             mammalian  
           
            49 

taaaaataat aataataata ataataataa tg                                   32 

 
           
             50  
             29  
             DNA  
             mammalian  
           
            50 

taaaaataat aataataata ataataatg                                       29 

 
           
             51  
             28  
             DNA  
             mammalian  
           
            51 

ctcctcagtc tataagtaac aataacta                                        28 

 
           
             52  
             28  
             DNA  
             mammalian  
           
            52 

ctcctcggtc tataagtaac gataacta                                        28 

 
           
             53  
             29  
             DNA  
             mammalian  
           
            53 

cgctcataag ggttaaaaac aacaacaac                                       29 

 
           
             54  
             29  
             DNA  
             mammalian  
           
            54 

cgctcataag agttaaaaac aacaacaac                                       29 

 
           
             55  
             30  
             DNA  
             mammalian  
           
            55 

tctccaagtg caaaaagaca taatcagcag                                      30 

 
           
             56  
             30  
             DNA  
             mammalian  
           
            56 

tctccaagtg cataaagaca taatcagcag                                      30 

 
           
             57  
             25  
             DNA  
             mammalian  
           
            57 

tatatatata taaaatgtgt ataca                                           25 

 
           
             58  
             29  
             DNA  
             mammalian  
           
            58 

tatatatata tatataaaat gtgtataca                                       29 

 
           
             59  
             19  
             DNA  
             mammalian  
           
            59 

agagcatgga ggacttgca                                                  19 

 
           
             60  
             19  
             DNA  
             mammalian  
           
            60 

agagcatggc ggacttgca                                                  19 

 
           
             61  
             19  
             DNA  
             mammalian  
           
            61 

ggaaggcgcc ccagatgta                                                  19 

 
           
             62  
             20  
             DNA  
             mammalian  
           
            62 

tcagacacat taaccttgca                                                 20 

 
           
             63  
             22  
             DNA  
             mammalian  
           
            63 

caatagacaa gtgatttcac tg                                              22 

 
           
             64  
             22  
             DNA  
             mammalian  
           
            64 

atgcttaatt ataactatat tc                                              22 

 
           
             65  
             20  
             DNA  
             mammalian  
           
            65 

ccataatagg ttcattgccc                                                 20 

 
           
             66  
             18  
             DNA  
             mammalian  
           
            66 

ggcttaaagg ggccaagt                                                   18 

 
           
             67  
             18  
             DNA  
             mammalian  
           
            67 

cccaccacca tcacctct                                                   18 

 
           
             68  
             20  
             DNA  
             mammalian  
           
            68 

atgttatctc attgcctttc                                                 20 

 
           
             69  
             20  
             DNA  
             mammalian  
           
            69 

gaagaaaggg gagaatcaag                                                 20 

 
           
             70  
             20  
             DNA  
             mammalian  
           
            70 

ggaaaatgca atgccatatc                                                 20 

 
           
             71  
             20  
             DNA  
             mammalian  
           
            71 

atcctgaatt tcctcaactg                                                 20 

 
           
             72  
             20  
             DNA  
             mammalian  
           
            72 

agagactgtc tgtatcccat                                                 20 

 
           
             73  
             19  
             DNA  
             mammalian  
           
            73 

aggcctgagc caggggtat                                                  19 

 
           
             74  
             20  
             DNA  
             mammalian  
           
            74 

ttctaagcca ttcgctcctg                                                 20 

 
           
             75  
             20  
             DNA  
             mammalian  
           
            75 

attctggacg tttcacctgc                                                 20 

 
           
             76  
             20  
             DNA  
             mammalian  
           
            76 

tacttctgct gacaccacta                                                 20 

 
           
             77  
             20  
             DNA  
             mammalian  
           
            77 

ggatgaagga gacatacact                                                 20 

 
           
             78  
             20  
             DNA  
             mammalian  
           
            78 

gccgttcaca agctcaagta                                                 20 

 
           
             79  
             20  
             DNA  
             mammalian  
           
            79 

ggacttcttt cttagaatat                                                 20 

 
           
             80  
             20  
             DNA  
             mammalian  
           
            80 

atcaaacctg acccacccaa                                                 20 

 
           
             81  
             20  
             DNA  
             mammalian  
           
            81 

atgtgatcct tctttgactg                                                 20 

 
           
             82  
             19  
             DNA  
             mammalian  
           
            82 

gaaaggccat gcacctaac                                                  19 

 
           
             83  
             20  
             DNA  
             mammalian  
           
            83 

tagctcatct tggagcgaat                                                 20 

 
           
             84  
             20  
             DNA  
             mammalian  
           
            84 

tttggaggaa aagtggaaga                                                 20 

 
           
             85  
             17  
             DNA  
             mammalian  
           
            85 

aatacgactc actatag                                                    17 

 
           
             86  
             20  
             DNA  
             mammalian  
           
            86 

gaattgcatg ctgggttctg                                                 20 

 
           
             87  
             20  
             DNA  
             mammalian  
           
            87 

ctgtggcttc cagaggttac                                                 20 

 
           
             88  
             20  
             DNA  
             mammalian  
           
            88 

taatgtggtc attggcaggt                                                 20 

 
           
             89  
             20  
             DNA  
             mammalian  
           
            89 

cagatgagtc cttgtgcccc                                                 20 

 
           
             90  
             18  
             DNA  
             mammalian  
           
            90 

acccgggcca gagcagcg                                                   18 

 
           
             91  
             20  
             DNA  
             mammalian  
           
            91 

gcggaataaa gatatctctc                                                 20 

 
           
             92  
             19  
             DNA  
             mammalian  
           
            92 

gatgagatga aggcaaagg                                                  19 

 
           
             93  
             20  
             DNA  
             mammalian  
           
            93 

tcaccaggga tgcttccagg                                                 20 

 
           
             94  
             18  
             DNA  
             mammalian  
           
            94 

ggttacccac attccatc                                                   18 

 
           
             95  
             20  
             DNA  
             mammalian  
           
            95 

cttatgccat ggatcatgtc                                                 20 

 
           
             96  
             20  
             DNA  
             mammalian  
           
            96 

aagtggttct gaaaccactg                                                 20 

 
           
             97  
             19  
             DNA  
             mammalian  
           
            97 

ccagagtgtc tgattcagc                                                  19 

 
           
             98  
             20  
             DNA  
             mammalian  
           
            98 

aaggcaagcc atctgataca                                                 20 

 
           
             99  
             20  
             DNA  
             mammalian  
           
            99 

aggccaacga tctaagcatg                                                 20 

 
           
             100  
             18  
             DNA  
             mammalian  
           
            100 

ggagtggcag aggcctgg                                                   18 

 
           
             101  
             19  
             DNA  
             mammalian  
           
            101 

caccatttct ccaggggca                                                  19 

 
           
             102  
             20  
             DNA  
             mammalian  
           
            102 

caacgaacca agactgtcat                                                 20 

 
           
             103  
             20  
             DNA  
             mammalian  
           
            103 

gttaattcat tacactcacc                                                 20 

 
           
             104  
             20  
             DNA  
             mammalian  
           
            104 

ataggtcact gagaggttgc                                                 20 

 
           
             105  
             20  
             DNA  
             mammalian  
           
            105 

agcttgttgt atccttccag                                                 20 

 
           
             106  
             20  
             DNA  
             mammalian  
           
            106 

tcatactcct tgttgtcccc                                                 20 

 
           
             107  
             20  
             DNA  
             mammalian  
           
            107 

cggctagctg taagatctga                                                 20 

 
           
             108  
             19  
             DNA  
             mammalian  
           
            108 

catggaacta agctgagcc                                                  19 

 
           
             109  
             20  
             DNA  
             mammalian  
           
            109 

cgcagaatgt cagggagaag                                                 20 

 
           
             110  
             18  
             DNA  
             mammalian  
           
            110 

tccaggtgca ctgagagt                                                   18 

 
           
             111  
             22  
             DNA  
             mammalian  
           
            111 

ttctagcaca attgccttgc ct                                              22 

 
           
             112  
             20  
             DNA  
             mammalian  
           
            112 

aacattccat acatcctggc                                                 20 

 
           
             113  
             20  
             DNA  
             mammalian  
           
            113 

gcaggaagac actgactttg                                                 20 

 
           
             114  
             20  
             DNA  
             mammalian  
           
            114 

gccttccaga cacttacggt                                                 20 

 
           
             115  
             17  
             DNA  
             mammalian  
           
            115 

attaaccctc actaaag                                                    17 

 
           
             116  
             24  
             DNA  
             mammalian  
           
            116 

gcccagagca aggtaagcac ttcc                                            24 

 
           
             117  
             24  
             DNA  
             mammalian  
           
            117 

cccttcttat agatgtgtca ccag                                            24 

 
           
             118  
             24  
             DNA  
             mammalian  
           
            118 

tgaagaaaga tggtaaccag cctc                                            24 

 
           
             119  
             24  
             DNA  
             mammalian  
           
            119 

tgtgcattcc agtttatgtc gtag                                            24 

 
           
             120  
             24  
             DNA  
             mammalian  
           
            120 

aggaccagaa aggtaattct atac                                            24 

 
           
             121  
             24  
             DNA  
             mammalian  
           
            121 

tttcaaatcc agaacccaaa aata                                            24 

 
           
             122  
             24  
             DNA  
             mammalian  
           
            122 

aagcagcaga gggtgagtga aact                                            24 

 
           
             123  
             24  
             DNA  
             mammalian  
           
            123 

ctttgacttc agctcttctg accc                                            24 

 
           
             124  
             24  
             DNA  
             mammalian  
           
            124 

tcagggacat cagtgagttt tgga                                            24 

 
           
             125  
             24  
             DNA  
             mammalian  
           
            125 

cctcttccac agtcaaacct gacc                                            24 

 
           
             126  
             24  
             DNA  
             mammalian  
           
            126 

aagagagaaa aggtaagaag tgat                                            24 

 
           
             127  
             24  
             DNA  
             mammalian  
           
            127 

tctcttttgc agaaagatag agtc                                            24 

 
           
             128  
             24  
             DNA  
             mammalian  
           
            128 

ccctgcagtt aggtgagcag gccc                                            24 

 
           
             129  
             24  
             DNA  
             mammalian  
           
            129 

attctcttcc aggttctgat ccag                                            24 

 
           
             130  
             18340  
             DNA  
             mammalian  
           
            130 

caatagacaa gtgatttcac tgcgggaaga caattcagag ccctgttcca ggctcctcac     60 

attgattctc tctgtcttct tccactcctc tttgtcatct ttgatgtccc cttgtgagct    120 

acgaaaagac tttctgggac acgacaggat aaaaaaataa ataagtgcaa gcagccattc    180 

attaaacgtt tagccaggat gctgctttaa ctgcatccca tcatatctca ttaatcttca    240 

caccagtcct gagatcaggt actattatta acccgatttt acagatgtga ggaactgagg    300 

cttaacgaag gtaagtaact tgcaggtgcg ggtatccagc tctctaactc cagagcccat    360 

gctcttaaaa ccctattact tgtccctggt ggaggtgaac actgggggcc ctttcatata    420 

ggactagccc tcgggctgca atctgagcgg aaaagggagg atgagggcat acttcgaagc    480 

ttcttttgca taactggcgc tggattttta ctgagacttt acgttacagt tttttttttt    540 

taattttcaa ggtgctttta cgaacacatg aataaaatat ttgtgtcatt ttgaacctta    600 

cttgtcttat tttatgcatg tatttattta tgggggggca caaggactca tctgtggtgg    660 

tgcagccact gtaaataaat tagtgaaact acttcacgtc aatttctgtt cagtacactt    720 

tagtgatgga tcggaggaaa ttaatacatg tttacaaaaa gcccctcccc cagttgttac    780 

atatgcctca gagataccag ttgtgaaaag tgcaggtgca cttacacaca tacgcacaca    840 

caccccacaa atggtatcat acgaaaaaac atacctgcaa tctgctttgt ccacttaatt    900 

gtatatcttg gatacagaac ttgtttcact ggaaggctaa aaggcaaagt ctggggaggc    960 

ctagaggaca caggggatgg gaggaggcgc tctgagctgg atgtaaggtc tccacccacg   1020 

gccagagcac aaggtcggat aaccagtggg cctgccggct tggctgcctg ggccctcccc   1080 

tgccgagaca aacggctgga gggaggaagt gtgcggctgg gaagctccgc tgctctggcc   1140 

cgggtttccc atttccccct tcccgcgctg agacggcgag gaaagttagc ccggaaatct   1200 

gcgcccgcct aaaacccggc ctggtcccag ccaccgcccc aggaacttcc cccaccgcag   1260 

gggcggaggt cgagagcagg gatggagaag tggacctgcg cgggtggact ccggggcgcg   1320 

ggtggactcc ggggcgcggg gggactccga ggagcgggtg gactgtgggg cgcgggtacc   1380 

gtctcgcagc gacctctgtc ggcggctctg gggatggccc gcatctgtct gcgtgtacct   1440 

ggtatacgtg caggtacatg ttcctgttca cgtgcagact gggcggggga tgggggggtc   1500 

cacaccggtg tacacctttg catacctctt agcaacttga aattccacca cgagagatat   1560 

ctttattccg ctattcctgt gcatctgcac ggagccccta gggccataga tttgtgtgca   1620 

aatgaaatga ggatgtagtc tgggtgccca agggggggtg ccttgagtgt ggttgtctgt   1680 

atgcctccct gagggtattt cactttctgc tcccatccgc ccctatgagc gagtacctat   1740 

gagcacagga tgtgcacata tttgagtctt attagtggta cacgcagttt tatcatctcc   1800 

ccaggtctgt gtctgtatga aatgtgcatg ggtgtgtgtg tgcacgcgtg tgttcccact   1860 

cggggaatgt ggggagaggt gcatggagcc aagatgggtg gtaaatagta tgtttctgaa   1920 

attaaaggac taatgtggag gaaggcgccc cagatgtact aaaccctttg cccttcatct   1980 

catcctctct gacttgggaa gaaccaggat tttgttttta agcccttggg catacagttg   2040 

ttccatcccg acatgaactc agcctcccgt ctgaccgccc cttggccttc cttcttcctc   2100 

gatctgtgga acccagggaa tctgcctagt gctgtctcca agcaccttgg ccatgatgta   2160 

aacccagaga aattagcatc tccatctcct tccttattcc ccacccaaaa gtcatttcct   2220 

cttagttcat tacctgggat tttgatgtct atgttccctc ctcgttattg atacacacac   2280 

agagagagac aaacaaaaaa ggaacttctt gaaattcccc cagaaggttt tgagagttgt   2340 

tttcaatgtt gcaacaagtc agtttctagt ttaagtttcc atcagaaagg agtagagtat   2400 

ataagttcca gtaccagcaa cagcagcaga agaaacaaca tctgtttcag ggccattgga   2460 

ctctccgtcc tgcccagagc aaggtaagca cttcccaagc ccctacctcc ctcccctccc   2520 

tgtgggcctg cagctgtcca ggtgtagaaa ccgttagtgt gctaccccag cagctggcag   2580 

gagggagttg gtggattcct ggaagcatcc ctggtgagtc atctgctgga acattagtga   2640 

aaacttagta ctctagggac cgatgtacag tgtccatttt aaaagccacc taataataac   2700 

tgtatagcaa gatctgtgtg tatgcatagt ttgtggaaat gtttgtttta tcttattttg   2760 

aagtggtgtg tattgatgta taaaagtata ttcctaaatg ttaatgccca tcagttaaag   2820 

gattgtatag agctaaagtg agtggtgcct gccttactat tgaaattttt aaaaagcctt   2880 

tcgtgcattc cttaaagtaa ttggattcat aattataata atgttacaaa tcagacttgc   2940 

tcccatattt gtgatggtct tggtcgtcag ttgtgatatc aaccaaaatg acagctggga   3000 

tccccattct tgtggattaa ctaactttgg ccccagttaa aaaatgaaaa gctattattg   3060 

cttcctaaag agtttttaaa tctgtgagaa gggggaaaaa aaggtttttt tacttgccca   3120 

ggtaaaattg tgtgcaacaa caatgtcatt ttaacaaagg gattactaaa ccccaggtga   3180 

tgacccactt tttcaaacaa ggaattgcaa gatactagat ggaatgtggg taacctctta   3240 

gagtttagtt tactggaatc tgaaattgta tgatttccgt acattccttc atgctactaa   3300 

taagtcatgg aatccctctg tgctggactc tgggggtaaa gtgcaaaaca agatagacat   3360 

gatccatggc ataagagagt tcactcagtg tggcagagaa gacaaacagt aaagaagaga   3420 

ctgtattgtt gccattgtag taaatgctgt ggaaggaggg gagcaaatag tgggtcctga   3480 

cttcatctag gcaccatgac acttcacgtg aattatgtca ctaactcctt accacagtag   3540 

catgcccatt ttatagatca ggagtgtggg gcttaaaggg gccaagtgac tcacccagag   3600 

tcacacagtt ctagaagtct gcctggccct caaactaggg atctttgcat gtgccaccca   3660 

tgccatctgt gactatatct ttttattctt taatagtaac tcccttttct aattaaaggt   3720 

aacaaacaaa acttaaaaaa gagatgccca cccaagcgtc attggcatgc tgatgttggc   3780 

accagtgttg ggaagccctt agcatactcc aggaagtagg agtgtgtaac gtggggtccc   3840 

tttgtccttc atgcaagggt ttcaagagtt tagaaaacct atgaaattgc acacacaaaa   3900 

atgtgtttta atcatcaaga ctctcagact taccatgctg agaaatgtgg gctgagggat   3960 

agtgctccct agtatcagct gatgggccag agagccaaag gaggagaccc accaccatca   4020 

cctctcctgg acagtgctct gtggtttcaa atgtaggtga tactaaaatg ggagttgttt   4080 

cttagaacca tggccggggt tcccttgacc ctgaagtgca gttcacctga gattgtcaaa   4140 

tgcatctgag gcatgcagag gaagtgctgg gcacacagct agtgggaata cctcagcgta   4200 

agtggccagg agatgccagg aatctccact atttcccttc cagtgtgcca gcctctgggt   4260 

tttacaggcg catgtaattg cagtacctct gtgcacattt ccctactgcc tagaatgact   4320 

ttcttgacta tccatgatat ataaaacaca gataccaaat tgttccctta cctcttcctc   4380 

taggttcaag ttaatatgtt attggttgcc ttctataata tgttatctca ttgcctttcc   4440 

caacaagtct ttgagataag tattaggtcc attttataga caaagagact gaggctcagc   4500 

gagtaacttg gccaacaagt tgctcccact gctcaacagc aaatgagcgg tgggaccaaa   4560 

attcaagcct gtgtctgttt aacttcaagc ctgtgaatgt actaaccggt gccctgtgcc   4620 

agctagtact ttgctacagt cataacctag actgaagtga tagccatgcc cctaaaactc   4680 

catgctgtgg tgacagcact gagcagtgtc caagaaggct tgacttctag gcctgtctct   4740 

gccactcaca aactttataa gggaataaag tacatagcaa ggtccgccta gagttagtag   4800 

cagttctgac aaagctgtaa tttgtcaata ttccgtcacc caacccagga atgctcattt   4860 

ttaaggtatt tgactgaaac agttgagcat tgcccttcat atagtttaaa acagtggttt   4920 

cagaaccact ttcctccaga ccatgggtgc tctgcaaggt gaatggagtt gtttcagaat   4980 

gtttcaataa tcatccctac ctcattcgta agtggcatgt aatttttgca atcggaagat   5040 

tttcataaac cctggatact aacctagact ggtttctata tcagatggtg gcttatttaa   5100 

cataaaatta tgcattttac tatttcatgg tggatatatc aatatgttgt ggtcttttcc   5160 

caatgaacac tttgattttc aggggttctg gaccctgaac atgggttaaa ccagtggttc   5220 

tcaaggtgtg gtcttagcgc cagcagcatc tgcttcccct ggaaactttc tagaaatgca   5280 

tattctcagg ccctcatgcc tgctgaatca gacactctgg gggtgggact cagccgtctg   5340 

ttgtagcagt gcttccaggt tatcctgaca gtcactcaaa ttttagaacc actaggttct   5400 

ctatatggga gagagtagtc tttgaacttg gaaaacaaga gaagctaaac ccctacagca   5460 

agggctggtg accaggtcgt tgccagaacc tgaaagttcg cctctgtatt accgttcctg   5520 

tccctaaccc aagtccttca gttctgggtg ctccagcaca cactgctttg tgctgcagtg   5580 

atacaaatgt atggctcatc tccccagctg gcggggaggc atttaacaca ctgacttaat   5640 

aaatatttat tgagtaaaag tatttgctcc taggaagcgg gatccaggta agcccttttt   5700 

ttctctctca actgcttcta gcccagtgct ctttatgtag taagcactaa ataaacaact   5760 

gctagatgtt gatccagaaa gtcacattcc ttctctaagc tttaagtttc tcatcttaaa   5820 

aataagagga ttgtatcaga tggcttgcct taggtctctt tcagctccag agccccaaat   5880 

accctatggt tctctattta gagatgttct tccccacaga ctgccataga actcctgtaa   5940 

tttacttagt atttgcttga cagtatggag aagaaagggg agaatcaaga ttttatttaa   6000 

aaaaaaagta gctagaatgt gtatatggtt cacaaaggta acaagaatta ttgacattct   6060 

ttcttctctt ttttcttcct cttccttctc ttttcctcct tctcttcccc ctgcttctct   6120 

cccttcttat agatgtgtca ccagcagttg gtcatctctt ggttttccct ggtttttctg   6180 

gcatctcccc tcgtggccat atgggaactg aagaaagatg gtaaccagcc tctcattatt   6240 

ctctgtggag gccccacttc taagccagga ctcttgggca gccactggtg ggaaatcaaa   6300 

ctgaaatggg caaccatgca ctgggtcctc tagagaaagc catcactctg ggaaaatgca   6360 

atgccatatc tctcttttct actttgatgg tatctatatt gtttggtttt cacattggat   6420 

gacattggta cactatggtg gggaaagaca tatgatatat gatatggtgg ggaaagacat   6480 

atgacatatg atattttcca atattactaa aaactgtttc acacaattaa aattccaaag   6540 

tagaggattt gcaaagtata acaactgtgt tcgtttctca ttccaccaca tgatactgcc   6600 

ccctcagttg gcactgtgat gacttacctc tgaccaagca ctttggagga agcataggat   6660 

tcagactcac attgacttgg gttcaagtcc taggtctgtc aatgactggt tatgtgactt   6720 

taagctgggt cacctctaat cctgaatttc ctcaactgta aactggatgt tacaaagtgg   6780 

atgcctacca cgtgggttat ttagtgggtt aatgaatgca gaatacaact cgacagatag   6840 

taaagtgaaa gtaaatgtca gctagtatta ctattttggt tgtttaaaat atctttcatg   6900 

attcaagaga tactttttat tatcccaatg atcagtaaaa attattagta gactaataga   6960 

atagttaatg gtaaaataag gagttctgcc catccttcta gtatctcaca ctcagtaaat   7020 

gtgcattctg accgttggct gtacctgaaa gaccctcaga tttttatcac tgaagccaac   7080 

atcataatgt tggcgattac tatctttaat tgtataataa taatagttaa tgtttattga   7140 

gtgcactgtc tcacttaatt ctcacaagag ccatatgaag tagagactgt ctgtatccca   7200 

ttttacagat gttggaaact gaggccagag agattaagta acttgcccaa tgtcacatac   7260 

ctggtaaggg tggaacaggg acttgatccc aattctgtct tgcttcaaag ctggtgcact   7320 

taaatttgtg aaaacgtttt tacaaggaca tgaagtaatt ttttcccagg tctttggaga   7380 

gctgaataag aggaaatgga cataaattaa ggatgaaaat atttcagctg atgatcagaa   7440 

ataatctttt gatattctag aaagtaccat cttgaaatgg gtacctgaaa gaaatctggg   7500 

accactcctc tcttctcaag aattttagga agacagaatc cagccacccc ttctccatga   7560 

gaatttgaga gatttagaca ctctcttaag taaaggcaaa ggcctgagcc aggggtatat   7620 

ggcagatccc ttccaaccct gggattgtta gcgagctcag gaaccttggt cctggcatat   7680 

ttgacccctt agtgacttct gatttggtaa accacagaaa ttccagaaaa tcagtgtgag   7740 

aaactctctg aggtgtgact taggagggca gacgatgcag tgaggctaag tgccaggttc   7800 

ttgatgctcc tcttcagctt tcctcctgca gctgttttcc ctgctgttga gcaaacatct   7860 

tctagggctt ccgagcctca gttgggacag gaaagtaacc atgctcttca ggtgtcaggg   7920 

ggacaaaaaa aaaaccaaga aaaagccaaa agtgccacat ggttttacat cagcacagct   7980 

aatcatttcc ccagagttgg accccaaatg cttttgacct cttatttttg ttatccattc   8040 

agtccttata atccaattga tgtaaagtga aaactttata ttctacaatg ctttacatcc   8100 

agaggccaat aacgagaacc accatttata aagcatgtaa gggcactgtg tatgagctta   8160 

tataatccac atatccacct tctaaagcaa gggctaatat ttttctcatt ttaaagatga   8220 

tgacactgag gcttacagta gttgaatgtc ttgccaaagg ccacaagact ggagcaaggg   8280 

ctagagctgc ttctaaatcc aggcctctgc cactccaaaa tgcaggctct caaccactgt   8340 

gactcataaa cttgagcagg catcagcacc atctggagag cttaagaacc atataactaa   8400 

atccatccca aggtttctga ttcagcagct gagaatttgc atttctgatc tattccaagg   8460 

tgatgctgct gatggtgttt catcgatcat gctttgggaa ctactacatt aaacaattct   8520 

attcaattaa taatttatgc atggattaaa aaaatgaatg aagctttgct atgacacact   8580 

ctgaaatact atactaagcc attcctcaaa ggccagttta gacactagca ttaggcatcc   8640 

cttgcaaagc ccaagagaca aaaggtctga gctatagccc ttgtacttct gacttgctgt   8700 

gaccatgctt agatcgttgg cctcagtacg cttcttcatt aaatgggaag actaagctct   8760 

actggactgc ttcataagag tgtaagatag ctaaaaataa taataataat aataataatg   8820 

cagagagaat gaaaatctcc actggtgatt taaaacagag ggaacgaaat ccttaaatat   8880 

ccatggaaaa ttgttaagag agtttctctg tacagttggc tgactcctcg gtctataagt   8940 

aacgataact aacaccgaat ttactgtgtg gcagacactg tgctaagtac tttacgtgct   9000 

tttttttttt ttcatttaat cctcagtcaa atgtaaggca gatactgtta ttattatcat   9060 

tttacagatg aggaaactga ggctcatgat aatgaaatac cttgttccaa atcccccagc   9120 

tggttagtgg agacaggatg acagtcttgg ttcgttgttc tcgacaccct gagcttttaa   9180 

ccactatgtt actctgctga atattgtgcc ctgccgtatt ctctatgaaa ctgaaattgt   9240 

gctggaagtt tctctccccc agacctttgg caaagagtct tgtgctgttt gcagtttttg   9300 

gtatattaag gtgtttccaa tctgctaaat aatcaaaggt tactattaaa ggcagccttc   9360 

cagtcaatga gtcgatggca gctataaaac tctttgtttc tcttttccat gaccttgagc   9420 

ccaagcaggg tctcatgcct tgagatcatc tcagcaagca tttgccaaat acttgttgta   9480 

aacaaggttg tgtttaggca atggggatgc ccgaagggtt aataaaacac agtcccagag   9540 

ttcctggagc ttacagcctg gttctccact ttatgtgcat tccagtttat gtcgtagaat   9600 

tggattggta tccggatgcc cctggagaaa tggtggtcct cacctgtgac acccctgaag   9660 

aagatggtat cacctggacc ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc   9720 

tgaccatcca agtcaaagag tttggagatg ctggccagta cacctgtcac aaaggaggcg   9780 

aggttctaag ccattcgctc ctgctgcttc acaaaaagga agatggaatt tggtccactg   9840 

atattttaaa ggaccagaaa ggtaattcta tacccttgga tagtatcaat tttctctttc   9900 

gctcataaga gttaaaaaca acaacaacaa caaattgaaa agccaagtca tggtgagtgt   9960 

aatgaattaa catcaagtct cttattgatg ttaattgatg ttaacctcca ttttcctttg  10020 

ctttcctgga ccctttgggt tatcaaccat caaaatctca tattaaggga gtttcatgat  10080 

cagtctgaat gcttagcctc atgttttctt taaataatgg tgatattatt taatggctaa  10140 

tggaaattaa ccgatagtgt atcactctgc actggggtga tagccttcaa aaaatgaatg  10200 

cctctgccag gcatgttagg tgtgtagtgt actctgcaga atcaacaccc cactgggata  10260 

ctcccaatcc ttatggagct acccaagagg caacgcatgg aagaacttca ccctgtacca  10320 

tctggtgatc tgtgattcat cacaatcaaa acctttctgc aaaaaactcc taaatattga  10380 

atttttgttt ttttcaaatc cagaacccaa aaataagacc tttctaagat gcgaggccaa  10440 

gaattattct ggacgtttca cctgctggtg gctgacgaca atcagtactg atttgacatt  10500 

cagtgtcaaa agcagcagag ggtgagtgaa actgctctgg tttctcagca tttttctaga  10560 

actatttcat taagaaatta agggcaacct ctcagtgacc tatcagttaa tgataatggg  10620 

aaaagcaaag tcaaacccgt gttttttcaa ccgcccttcc ttgtctacat tgaagaaaga  10680 

acatggagat tttagccgat tgcttgaata aatgtatgtg ttggggcagg atattattgg  10740 

gaactgagaa tagtctctgc tgtgtttgaa cccactcatc caaattgcct ggccatgctt  10800 

cctgaagcct catagcacca aagaaaggga taaaaggaga attcaaagct acaaatgact  10860 

tgctgaaatt gcaccttgag tcaaaaataa aaacaagagc tccagggcgt agatcttagg  10920 

ggccctgaag cagactccaa aactcgatga ggcctcccga aattttccca gggccacctc  10980 

aactcctttt acttctgctg acaccactaa tctgaagttc gctgttggtc caatgcacct  11040 

ggactttccg taagaaagca acttccataa atacaagacc tatgtgttaa cccccatgtg  11100 

gcttacttta atcatcaccg aagcaaaccc caggtgatca tcctgacttt accattattt  11160 

cactgagtaa attaagcatt ggggtctcac tttttcatct ttaaaaggaa aatgcttact  11220 

aaagaaatgt ttctccaagt gcataaagac ataatcagca gaggaatggt taaataaaac  11280 

atggtacact atactcttgc ttaatgtgca gtcattgaag tggataaccc aacccatatg  11340 

ttttgtcatg gagagctccc cataatatgt tcagagggga aaaggatggt tacataatca  11400 

tatgtataca atttgatcct agttcataaa aataaaatct atatgtataa gtaaaatata  11460 

tatagtggat atatataatg tagagatgta tataacatgg attatatata taatgtgtgt  11520 

atacatatgt gtgtgtgtgt gtgtgtatat atatatatat atatataaaa tgtgtataca  11580 

attatcttga atattcattg aaaaagttct ggccaggcac agtggctcac acctgaaatc  11640 

ctaactcttt gggaggctga gacagaatga ttgcttgagg ccaggagttc aagaccagcc  11700 

taggcaacac agtgagaccc catctcagaa aatattaaaa ataaaaaaat taggtgggtg  11760 

tggtggcaca cacctgtagt cccagctact tgggaggcag agggagggga tcacttgagc  11820 

ctaggagttt gaggctacag tggggtctga ttccaccact tcactccagc ctgggtggca  11880 

gagcaaaacc ctgtctctta aaaaaaaaaa gagagagaga gagagaaaga aaaagaaaaa  11940 

ggaaggtctg gaaggataca acaagctatt attagtactt aaacctgtgg agagcagtta  12000 

aggatgaagg agacatacac ttctttcctt tatatggatc tttatcatct ttacttttat  12060 

aattagtgtg tactgatttg tgtattgatt ttataattaa aatgggaaaa aatgaattta  12120 

agtttttaac aagggggttt aataatcaga gattctagat ctaaaacaaa caaaaacttc  12180 

catattcatt tagtccagag acatgtaagt gctcttgaat ttaagctttt tctcctgggg  12240 

agggcagttt cttaccctct gggtagaaat cagcccagtt ggagaaactg tgtcctcaga  12300 

caacagttga ggccttacct gccttactgg ctacaatcac taggaactct ctccccaatg  12360 

tgtaacacag gctaatttct gtctttgact tcagctcttc tgacccccaa ggggtgacgt  12420 

gcggagctgc tacactctct gcagagagag tcagagggga caacaaggag tatgagtact  12480 

cagtggagtg ccaggaggac agtgcctgcc cagctgctga ggagagtctg cccattgagg  12540 

tcatggtgga tgccgttcac aagctcaagt atgaaaacta caccagcagc ttcttcatca  12600 

gggacatcag tgagttttgg atgattatat gtgctccata aggaaagata ctatttgtca  12660 

cgtgttcaca atgccccatg cactgtgggg taggtggttg acaagcatca tctcttttat  12720 

tctgcatcca aaaacaaaat acgatgtaga tactgttatc tgcattttaa ggaagaggaa  12780 

attgagtctt agaaaagtta agcaacttgc cccagatctc agatcttaca gctagccgtt  12840 

caaatccaga tccactccac tacagctgct ctttactgca ctttgattca gctgccagat  12900 

agtttccatg atgaatccca gagcctaatc aagcataata ttcatattca gaaccagggc  12960 

ttccttacta atggcaatta ttcccaacca atccttcctt agcatttgaa aagggacttc  13020 

tttcttagaa tataaaccct tccaaaatgg acatcttttt ttttaattgg cagataggga  13080 

tttcaccata agtcatttcc tttactattt attcattgac caggcagcat gataaagtgt  13140 

aatagaacca gagaacttgc ttcaaaactt atggagggtt tgtacttggt gggtggggtc  13200 

tagttcacat agggtggcca aggaaggcct ctctgaggag gtgacattta gctgacacca  13260 

aaaggaaaga tgtcagttgt gttaagagca gagggaagca tatgtgcgaa gcacctgcta  13320 

ggagccgtga tctttgtgtg gagcagtgcc aggcctacag agcccaacca cacaccctag  13380 

catgtctctg cctcctctta tctagaagac ctaattgagg aaggagtctt tgtgaaactc  13440 

actgctgtat ccttcatgca cagtccagtg gctggaacat aatgggcgct cagtattcat  13500 

ggaataaaca agcaaattga gcatagagac aattgactgt aactgctcca agacatgtcc  13560 

gcaccaaaag ctatgaaaag acaaaagaaa gggcagtaaa tagaaaatct atcatctcat  13620 

ccccagggag aggctcagct tagttccatg ttcagtgcaa agtgagggat tagcacagac  13680 

agggtggtcc ttcaatgcat ggcccataac cattaaagca gaggtcttct cactgtgcgg  13740 

tcccatctga ttgttcagtg atgaggattc tgagcatctc tcagatcctg caatacatgt  13800 

ggatctgaga tgtggccatt gataatgact gccttcccga ggcaccagcg tgagcacctg  13860 

cggcagaggt gcctcacatt tgccagccag gtgctcacag aagttaagta actatccagt  13920 

ggactcacag ctgatcaaag gtgcaagtga gatcataagc caaaaccact gaactccaaa  13980 

gccttattag gaaaataaag catgtttatc ctcttccaca gtcaaacctg acccacccaa  14040 

gaacttgcag ctgaagccat taaagaattc tcggcaggtg gaggtcagct gggagtaccc  14100 

tgacacctgg agtactccac attcctactt ctccctgaca ttctgcgttc aggtccaggg  14160 

caagagcaag agagaaaagg taagaagtga ttcaggtgca gtatattcct tggtcagttt  14220 

tacggaggcc caccataaag tgagaagatg aatgatgata ataacaatga catccatgta  14280 

tcacttaaca acagggatac attctgagaa attcatcttt aggcagctat atcattgtgc  14340 

aaacatatat ggtgtaccca cacaaaccta gatggtatag cctactacgc ttctaggctt  14400 

tatggtatag cctattgctc ctaggctgca aacctgtaca gcatgttcct gtactgaata  14460 

ctgtaggcaa ttacaacaca atggtttgta tatctaaaca gaaaagatat agcaaaaata  14520 

caatattata acaatatagg accactggtc atatatgtga tccttctttg actgaaatgt  14580 

tattatgtga tgcataatta cttttcttag cacttttcta tgtgtctaga gctgtgccaa  14640 

gggttttcca tgtttatttc acttaatcta caaaaattaa cgcaacaaag gtagctgatg  14700 

ttattcttgt ttttttaccc ccttttttgt ggaaaagagg ctttcctttt ttccagaaac  14760 

tgtggcaagg taaagtaaag ctgtagctga tgcaggaatt ttgtgtaggt gttagcagca  14820 

ctgccctcac tacgtgctca ttggacagta gcccaacccc aagaaaagga tggttggtag  14880 

ccagtagtat tatcatcatt tcacaagtga ttgaagactc agagaggtta agtgacttta  14940 

ccaaggtcac ccagctagga aatgacataa ccaagacata aactcaatct gccagacaga  15000 

aaggccatgc acctaaccac tccactacct ctgatgttgg tcattgatct tggcactcag  15060 

aattagtcct gatagaggag acctgggctc cagaagccta aaattgttgt ttcaactgag  15120 

tgcatgtaat gaatgataga acaggcaaga gatatcgccc ccaaaatgga tagctcctgg  15180 

ctgttccaga tattataaaa ttattttact aaacagaatg tctacactta tagaggctaa  15240 

gatattggct tcccagcttc ctcgccttac agcagaattc ctttgcctgt tgcaaggttc  15300 

cagaggccct tttgtaccgc cccagactcc tttcacccca cttttaaaat cactggacaa  15360 

agccctaatt cagcatagca tttagcatgt ggtagaaatt cagtgagcta gttactctct  15420 

gggaaaataa ttaggtaggg aggctatcct ggaatagata tttacctaaa tattatttta  15480 

catcttggca agtactttcc ctatttaaga tctgtatgac taataggtga tattgagtgc  15540 

ttcctatgtg ctaaagactt gctaagagtt tgacgtgatt tttaccttga actataattc  15600 

tatgaagtag gcattattgt tatccctatt tataagtgag gaaacagaca cagagaacct  15660 

aagacatttt cctgaagtta cacagctatt aagtagcagt gccagaattt gaaggcaagt  15720 

tttctgatga aatgatcagg atatggtatt tctcaatatc tcagggatgg ctagagcaaa  15780 

tctgtctctc tctcaccatc agctcaggac tgggtgagtg gccatggggt cttgaggcaa  15840 

ggcaattgtg ctagaaagat gaaagctggg ccaaacgatt tctccctcaa gggcttacaa  15900 

agtacaaaag ctgcacctac atgtggagtg tctgccagta ggtggtgcaa gttctatgca  15960 

cacccctgtg aattgcaagc acagtgccct aagaccaaga tgggcttgtt ttgggagagt  16020 

atgcattgca gaaacaggct cagcttaccc tgtgactatg ttgccaaggg gtcttcacag  16080 

ctttccttct cttttgcaga aagatagagt cttcacggac aagacctcag ccacggtcat  16140 

ctgccgcaaa aatgccagca ttagcgtgcg ggcccaggac cgctactata gctcatcttg  16200 

gagcgaatgg gcatctgtgc cctgcagtta ggtgagcagg ccctcaaagg ccagcccagg  16260 

cctgcactct cagtgcacct ggatgcaggg atatgattgg gggctgtgtt ggagaggaaa  16320 

gggggatgga gtggccagca cccagttgcc agaatcagaa acatacattt attcactaac  16380 

agatatttat ttggtgcctt tgttatgtag gacactgtgc tggccacagg gatattgcag  16440 

gaaagaaaac agaccggggt tctggcctcc taaagagaaa ggcaaagaaa agagagaggt  16500 

agccaggagg cagagcatgg cggacttgca agcttgcagg actcagaatc ttgttctggg  16560 

ggccccgggc cctgaaaccc actgaagggt tttcagcaag gaagtaacac aatcagatat  16620 

tattttaaga aaaccctcaa gaaagcctct ggcaagcatg gtgccagcca aattccaggc  16680 

cacataagga aggcctgggc cttctggcat gaaatccctg aaacccagtt gcccaggatc  16740 

atatgttgtg agaaataaga agagacattg ctgttacaat gtcaccccac atcaactttt  16800 

ggcattctct tccaggttct gatccaggat gaaaatttgg aggaaaagtg gaagatatta  16860 

agcaaaatgt ttaaagacac aacggaatag acccaaaaag ataatttcta tctgatttgc  16920 

tttaaaacgt ttttttagga tcacaatgat atctttgctg tatttgtata gttagatgct  16980 

aaatgctcat tgaaacaatc agctaattta tgtatagatt ttccagctct caagttgcca  17040 

tgggccttca tgctatttaa atatttaagt aatttatgta tttattagta tattactgtt  17100 

atttaacgtt tgtctgccag gatgtatgga atgtttcata ctcttatgac ctgatccatc  17160 

aggatcagtc cctattatgc aaaatgtgaa tttaatttta tttgtactga caacttttca  17220 

agcaaggctg caagtacatc agttttatga caatcaggaa gaatgcagtg ttctgatacc  17280 

agtgccatca tacacttgtg atggatggga acgcaagaga tacttacatg gaaacctgac  17340 

aatgcaaacc tgttgagaag atccaggaga acaagatgct agttcccatg tctgtgaaga  17400 

cttcctggag atggtgttga taaagcaatt tagggccact tacacttcta agcaagttta  17460 

atctttggat gcctgaattt taaaagggct agaaaaaaat gattgaccag cctgggaaac  17520 

ataacaagac cccgtctcta caaaaaaaat ttaaaattag ccaggcgtgg tggctcatgc  17580 

ttgtggtccc agctgttcag gaggatgagg caggaggatc tcttgagccc aggaggtcaa  17640 

ggctatggtg agccgtgatt gtgccactgc ataccagcct aggtgacaga atgagaccct  17700 

gtctcaaaaa aaaaaatgat tgaaattaaa attcagcttt agcttccatg gcagtcctca  17760 

cccccacctc tctaaaagac acaggaggat gacacagaaa caccgtaagt gtctggaagg  17820 

caaaaagatc ttaagattca agagagagga caagtagtta tggctaagga catgaaattg  17880 

tcagaatggc aggtggcttc ttaacagcca tgtgagaagc agacagatgc aaagaaaatc  17940 

tggaatccct ttctcattag catgaatgaa cctgatacac aattatgacc agaaaatatg  18000 

gctccatgaa ggtgctactt ttaagtaatg tatgtgcgct ctgtaaagtg attacatttg  18060 

tttcctgttt gtttatttat ttatttattt ttgcattctg aggctgaact aataaaaact  18120 

cttctttgta atcatatttt gggcattctc agctgatttg agtactctgc ttgcaagtct  18180 

accaggggtg cattttttcc cctatcattt aagcattgcc ctttcttgaa gtgatcccat  18240 

tccaaatttt gcagagttgc tctttcccct tatagtattt ccaaagtcag tgtcttcctg  18300 

agctcagggg atgtccctgt aatctgacaa ggaaggatcc                        18340 

 
           
             131  
             1263  
             DNA  
             mammalian  
           
            131 

atttgtattg tttgaagttt ttacaatagc atgtaatttt ctaagatttt aatttttata     60 

agtacacatg gcttctcctt tatttgaaat gtgtactaga tgatcaaagc atatgcatgc    120 

atgtattgct ttcttctagg agaaaataag tttttgtggc caaaaaaatt ctttaagtta    180 

tttttggttt ttagggggtt gcctccatgt cacagcttaa tcatcagaca cattaacctt    240 

gcagctcagc acgccctctg tttgtcagca gaccttcctc gcccataggg taagcaatag    300 

aaagcttata ggtatcagtt tattttgcct gggatcaggg tctggattgg gaagtgggac    360 

atgttgataa acctcttctc caaaattagg tcaatgggca tttggctcat attaccagaa    420 

tgctggctgg ccatgtacag cctgtctccg agagaggctc taatgtggcc cccacattag    480 

aacaacctgc caatgaccac attagaacct ccattgttaa aatgcaggtt cctgagcccc    540 

atcccagatc tgaatcacaa tctccaagca tcagccccaa gaacctgaat tttgttgtta    600 

catgcagata aagtacgaga accacttcct ccatgggtga actgaactta ccaaaatagt    660 

cagtcccgag gggcagagat ggcgtaggtg ccagttcttc tttctcatcc tagatgctca    720 

gagtcaaatt cttggctcag caatagacaa gtgatttcac tgcgggaaga caattcagag    780 

ccctgttcca ggctcctcac attggatctc tctgtcttct tccactcctc tttgtcatct    840 

ttgatgtccc cttgtgagct acgaaaagac tttctgggac acgacaggat aaaaaaataa    900 

ataagtgcaa gctgccattc attaaacgtt tagccaggat gctgctttaa ctgcatccca    960 

tcatatctca ttaatcttca caccagtcct gagatcaggt actattatta acccgatttt   1020 

acagatgtga ggaactgagg cttaacgaag gtaagtaact tgcaggtgcg ggtatccagc   1080 

tctctaactc cagagcccat gctcttaaaa ccctattact tgtccctggt gggaggtgaa   1140 

cactgggggc cctttcatat aggactagcc ctcgggctgc aatctgagcg gaaaagggag   1200 

gatgaggggc atacttcgaa gcttcttttg cataactggc gctgggattt ttactgagac   1260 

ttt                                                                 1263 

 
           
             132  
             49  
             DNA  
             mammalian  
           
            132 

tgtacagcct gtctccgaga gaggctctaa tgtggccccc acattagaa                 49 

 
           
             133  
             45  
             DNA  
             mammalian  
           
            133 

tgtacagcct gtctccgaga gagggctgtg gcccccacat tagaa                     45 

 
           
             134  
             20  
             DNA  
             mammalian  
           
            134 

tagctcatct tggagcgaat                                                 20 

 
           
             135  
             20  
             DNA  
             mammalian  
           
            135 

aacattccat acatcctggc                                                 20 

 
           
             136  
             20  
             DNA  
             mammalian  
           
            136 

gccaggatgt atggaatgtt                                                 20 

 
           
             137  
             20  
             DNA  
             mammalian  
           
            137 

gggtaagcga ttcaaacatt                                                 20 

 
           
             138  
             21  
             DNA  
             mammalian  
           
            138 

ggtattgcat tgtaggcaca t                                               21 

 
           
             139  
             19  
             DNA  
             mammalian  
           
            139 

gggcaacaag agtgaaact                                                  19 

 
           
             140  
             20  
             DNA  
             mammalian  
           
            140 

tcaaaagagg tccgtctaaa                                                 20