CHROMOSOME 21 GENE MARKER, COMPOSITIONS AND METHODS USING SAME

Abstract: The present invention provides isolated nucleic acids encoding human EHOC-1 protein and isolated receptor proteins encoded thereby. Further provided are vectors containing invention nucleic acids, probes that hybridize thereto, host cells transformed therewith, antisense oligonucleotides thereto and compositions containing, antibodies that specifically bind to invention polypeptides and compositions containing, as well as transgenic non-human mammals that express the invention protein.

Description: CHROMOSOME 21 GENE MARKER, COMPOSITIONS AND METHODS USING SAME BACKGROUND OF THE INVENTION A major endeavor in molecular genetics has been made in generating maps of the human genome. Human genome mapping consists, generally, of ordering genomic DNA fragments on their chromosomes using several methods, such as fluorescent in situ hybridization (FISH) , somatic cell hybrid analysis or random clone fingerprinting. DNA fragments that correspond to marked polymorphic sites can be ordered by genetic linkage analysis. Distances between polymorphic loci are estimated by meiotic recombination frequencies. High resolution maps based upon the estimated distances, however, cannot be constructed easily using such methods because the resolution is low at the molecular level and recombination frequency is not linearly correlated with physical distance. Thus, various obstacles such as, for example, the difficulty in obtaining highly informative markers and the paucity of identified markers that are evenly spaced along the chromosome are significant weaknesses of the currently available genetic maps. Most of the mapped markers are restriction fragment length polymorphisms (RFLPs) assayed by DNA hybridization. Although maps based on these markers have contributed greatly to the primary mapping of a number of diseases, they are still insufficient for many applications such as mapping rare monofactorial diseases, refining linkage intervals to distances suited for gene identification, and mapping of loci contributing to complex traits. Genetic linkage mapping is an important technology applied to the study of human biology and, in particular, for the delineation of the molecular basis of disease. Indeed, one of the most commonly used strategies 6/15144 PCIYUS95/14641 for studying human inherited diseases is by cloning the responsible gene based on chromosomal location. Genetic linkage maps, therefore, facilitate the identification and mapping of genes involved in onogenic diseases, genes involved in ultifactorial disorders, and are useful in carrier detection and prenatal diagnosis of hereditary disorders. A detailed linkage map is also a prerequisite for clone-based physical mapping and DNA sequencing of the entire chromosome. Human chromosome 21 is a paradigm for large-scale human genome mapping efforts. The smallest human chromosome, chromosome 21 has approximately 50 egabases (Mb) of DNA. Less than 1% of the 2000 genes estimated to be on chromosome 21 are known. A high resolution map of chromosome is of particular interest because of its apparent role in familial Alzheimer disease (FAD), Down's syndrome (DS) , amyotrophic lateral sclerosis (ALS) , and Finnish progressive myoclonus epilepsy (PME) . A gene defect responsible for FAD has been localized to chromosome 21 on the basis of genetic linkage to three pericentromeric loci. The gene encoding the precursor of the Alzheimer- associated amyloid β protein (APP) , the principle component of the senile plaques and cerebrovascular amyloid deposits of Alzheimer disease (AD) , has also been mapped to chromosome 21. The process of developing such a long-range contig map involves the identification and localization of landmarks in cloned genetic fragments. When there are enough landmarks for the size of the cloned fragments, contigs are formed, and the landmarks are simultaneously ordered. Currently, YACs, or yeast artificial chromosomes, are utilized for most mapping of the human genome. YACs permit cloning of fragments of > about 500 Kb. However, some difficulties have been encountered with the manipulation of YAC libraries. For example, in various YAC libraries, a fraction of the clones result from co-cloning events, i.e., they include in a single clone noncontiguous DNA fragments. A high percentage of YAC clones, particularly clones having high molecular weight inserts, are chimeric. Chimeric clones map to multiple sites on the chromosome and, thus, hamper the progress of mapping and analysis. Another problem endemic to YAC cloning is caused by DNA segments that are unclonable or unstable and tend to rearrange and delete. Bacteria Artificial Chromosomes (BACs) , provide an alternative to the YAC system. BACs mitigate the most problematic aspects of YACs such as, for example the high rate of chimeris and clonal instability. BACs are based on the E. coli single-copy plasmid F factor and are capable of faithful propagation of DNA fragments greater than about 300 Kb in size. BACs have a number of physical properties that make them amenable to physical mapping, including easy manipulation and an absence of chimerism. The lack of chimerism and the capacity to propagate large exogenous insert DNAs make the BACs excellent candidates for chromosome walking and the generation of contiguous physical maps. The need for molecular description of chromosome 21 derives directly from the association with several human genetic diseases. A map of contiguous units (contigs) covering this chromosome will speed the identification of the cause of these diseases. Indeed, a detailed map would provide immediate access to the genomic segment, including any pathological locus, as soon as it has been localized by genetic linkage or cytogenetic analysis. Thus, a need exists for identifying, characterizing, and mapping the numerous genes that occupy loci on chromosome 21, which will expedite the rapid translation of high resolution chromosome maps into biological, medical and diagnostic applications. The present invention satisfies this need and provides related advantages as well. SUMMARY OF THE INVENTION The present invention provides isolated nucleic acids encoding human EHOC-1 protein and isolated receptor proteins encoded thereby. Further provided are vectors containing invention nucleic acids, probes that hybridize thereto, host cells transformed therewith, antisense oligonucleotides thereto and compositions containing, antibodies that specifically bind to invention polypeptides and compositions containing, as well as transgenic non- human mammals that express the invention protein. BRIEF DESCRIPTION OF THE FIGURES Figure l shows a physical map for the consensus region for HPE1. Figure 2 shows a physical map for the consensus region for EPM1 in relation to the consensus region for HPE1. The locations of YAC clones, BAC clones and EHOC-1 were indicated by thick bars. Figure 3A shows the regions in which EHOC-1 has homologies to transmembrane proteins. Region 1 represents 29.4% identity in a 34 amino acid overlap with rat sodium channel protein III. Region 2 represents 20.4% identity in a 103 amino acid overlap with phosphoglycerate transport system regulatory protein of Salmonella typhimurin . Region 3 represents 29.1% identity in a 55 amino acid overlap with pyrophosphate-energized vacuolar membrane proton pump of Arabidopsis thaliana . Region 4 represents 24.0% identity in a 50 amino acid overlap with myosin-like protein of Saccharomyceε cerevisiae . Region 5 represents 17.9% identity in a 39 amino acid overlap with rabbit cardiac muscle ryanodine receptor. Region 6 represents 21.0% identity in a 62 amino acid overlap with rat cardiac muscle sodium channel protein alpha subunit. Region 7 represents 40.7% identity in a 27 amino acid overlap with rat skeletal muscle sodium channel protein alpha subunit. Region 8 represents a 30.3% identity in a 33 amino acid overlap with dystrophin cysteine-rich domain. Figure 3B shows a comparison of genetic distances to the EPMl locus in centiMorgans as computed by linkage disequilibrium studies. (Lehesjoki et al., Hum . Mol . Genet . 2:1229-1234 (1993)). Figure 4 shows the putative locations for splice variants and poly(A)+ attachments sites for different isoforms of cDNA encoding EHOC-1 polypeptide. DETAILED DESCRIPTION OF THE INVENTION Progressive myoclonus epilepsies (PMEs) are a heterogenous group of diseases which are characterized by myoclonus, epileptic seizures and progressive neurological deterioration including ataxia and dementia Berkovic et al.. New Engl . J . Med . 3.15:296-305 (1986). PME of Unverricht-Lundborg type (EPMl) is an autosomal recessive disorder with frequent consanguinity in Finland and Mediterranean regions with the incidence of at least 1: 20,000 in Finland. Genetic linkage analysis revealed that the locus for EPMl is on chromosome 21q22.3 Malafosse et al.. Lancet 339:1080-1081 (1992) and excluded Lafora disease from this region which is also a member of PME Lehesjoki et al.. Neurology 2:1545-1550 (1992). Linkage disequilibrium analysis made it possible to narrow down the candidate region to 300kb spanning the loci of PFKL, D21S25 and D21S154 Lehesjoki et al., Hum . Mol . Genet . 2:1229-1234 (1993); Lehesjoki et al.. Human Genetics 9_3:668-674 (1994). Autoimmune polyglandular disease type I (APECED) was also mapped to chromosome 21q22.3 by linkage disequilibrium analysis Aaltonen, J., et al.. Nature Genet . :83-87 (1994). APECED is an autosomal recessive disease resulting in a variable combination of failure of the parathyroid glands, adrenal cortex, gonads pancreatic β cells, thyroid gland and gastric parietal cells. Additional affects of APECED include alopecia, vitiligo, hepatitis, chronic mucocutaneous candidiasis, dystrophy of the dental enamel and nails and keratopathy. APECED usually manifests itself in childhood, but tissue specific symptoms may appear throughout adulthood. The APECED locus maps within 500 kb of D21S49 and D21S171. Holoprosencephaly is characterized by impaired cleavage of the embryonic forebrain and incomplete mid- facial development that manifest as a wide range of midfacial anomalies including cyclopia, ethmocephaly cebocephaly, premaxillary agenesis, hypotelorism, and a single maxillary central incisor. The most commonly associated chromosomal abnormality includes dup(3p), del(7q), deletions of chromosome 13, trisomy 13, triso y 18, and triploidy (Munke, AM J Med Genet 34:237-245 (1989)). The etiology is heterogeneous and may include aneuploidies for chromosomes 2, 3, 7, 13, 18 and 21. In order to narrow down the candidate region for HPE1, the deletion of 21(q22.3) was characterized in two HP patients by fluorescence in situ hybridization and quantitative Southern blot dosage analysis. For the smaller deletion, the regions for D21S25, D21S154, D21S171 and D21S44 were deleted and for D21S42 and D21S49 were not. Combining these data with previous reports of deletion of 21q22.3 (D21S112-ter) without the holoprosencephaly phenotype indicate that the region responsible for holoprosencephaly spans the l-2Mb region including PFKL and ITGB2 (CD18) . Four cases of holoprosencephaly with chromosome 21 anomalies have been published. Estabrooks et al. describe a minute deletion of chromosome 21(q22.3) (Estabrooks et al., AM J Med Genet, 36:306-309 (1990)) suggesting this region as a locus for holoprosencephaly (HPE1) . Described in the instant specification is the construction of the BAC (Bacterial Artificial Chromosome) Shizuya et al., Proc . Natl . Acad . Sci . USA 89:8794-8797 (1992) contig of this EPM1-APECED-HPE1 candidate region and the isolation of a novel gene from this contiguous map unit using a direct cDNA selection technique. In order to isolate genes responsible for these diseases, a cDNA library from a 14-week trisomy 21 fetal brain was constructed using Uni-Zap XR (Stratagene, La Jolla, CA) . More than 95% of the clones have inserts ranging from l-4kb (avg. 2kb) . In addition, a direct cDNA selection method was applied to BACs (Bacterial Artificial Chromosomes) in the 21q22.3 region. Sau3AI linkers were attached to cDNA that was synthesized from a trisomy 21 fetal brain. After digestion with Sau3AI, a second pair of linkers were attached to the cDNA which was then hybridized to biotinylated BAC DNAs which covered the candidate region. cDNA/BAC DNA hybrid molecules were captured on streptavidin coated magnetic beads, non-specific cDNAs were washed out, and specifically hybridized cDNAs were eluted and subsequently amplified by PCR. Twice selected PCR products were subcloned and analyzed. Southern blot analysis revealed that 21 out of 30 (70%) of the fragments yielded unique bands of the original BACs. Using these fragments as probes, cDNAs (3kb, 4kb and 5kb) were isolated from the library. The 5kb cDNA subclone (EHOC-1) maps proximal to neighboring D21S25 and exhibited homology to other transmembrane genes. The loci of these genes all map within the consensus region where holoprosencephaly, EPMl and APECED are localized. DNA sequence analysis of the 5kb cDNA showed a complete coding sequence of 3570 bp which revealed amino acid sequence homology with other transmembrane proteins including three sodium channel proteins. (SEQ ID NOS: 1-2; Figure 3) . Five BAC clones were isolated from the total human genomic DNA BAC library (Shizuya et al., Proc . Natl . Acad. Sci . USA .89.8794-8797 (1992)) by PCR screening using sequence tagged sites (STSs) containing PFKL, D21S25, D21S154 and CD18. Physical maps of the HPE1-EPMl-APECED consensus region with these BAC clones and YAC clones (Chu akov et al., Nature 359:380-387 (1992)) are shown in Figures 1 and 2. BAC-1 (230kb) and BAC-2 (210kb) were positive for D21S25. BAC-3 (170kb) was positive for D21S25 and PFKL. Agarose gel electrophoresis of EcoRI-digested BAC DNAs and Southern blot analysis showed that these 3 BACs are overlapping. BAC-4 was identical to BAC-3. BAC-5 (lOOkb) was positive for CD18. Direct cDNA selection was performed on 5 BAC DNAs (four of which were overlapping) which span the consensus region. EcoRI digestion of subclone DNAs revealed that 10% of the clones were chimeric. The average size of the inserts of non-chimeric clones was 400bp. Forty non- chimeric subclones of selected cDNAs were analyzed by using EcoRI-digested BAC DNA Southern blots. Twenty-eight clones (70%) showed unique signals on the BAC blots, 6 clones (15%) showed repetitive, and 6 clones (15%) did not show any signal on these blots. Using insert DNAs of these subclones as probes, a trisomy 21 fetal brain cDNA library was screened. Three overlapping cDNAs (3kb, 4kb and 5kb) containing poly (A) * tails were isolated and designated EHOC-1. The three overlapping EHOC-1 cDNA subclones were used for Southern blot analysis using EcoRI-digested BAC DNA blots. Only BAC-l showed unique multiple band signals indicating that these cDNAs originated from BAC-l. Identical sizes of the signal bands indicated that these clones overlap each other. Complete sequence analysis of the EHOC-1 5kb cDNA clone and partial sequence analysis of 3kb and 4kb clones showed that the entire sequence of the 3kb clone and part of the sequence of 4kb clone are contained in the 5kb clone. The 3 1 end of the 4kb clone, however, was different from that of the 5kb clone which indicated the existence of splice variants of EHOC-1 cDNAs. Upon further characterization of a total of 9 human cDNA clones (2.7kb-7.2kb) , alternative splicing and/or multiple poly(A) attachment sites was evident (See Figure 4) . For example, SEQ ID NO:4 contains an additional 1801 nucleotides of 3' untranslated EHOC-1 cDNA, which extends (in some isoforms of the EHOC-1 mRNA/cDNA) immediately beyond nucleotide 5107 of SEQ ID NO:l . In addition to a poly(A)+ attachment site beginning immediately after nucleotide 5107 of SEQ ID NO:l, six other poly(A)+ attachments sites have been elucidated. These six attachment sites occur at nucleotide positions of SEQ ID NO:4 corresponding to nucleotides: 10, 310, 1149, 1155, 1798 and 1801 (see Figure 4, clones cDF7, cEH18/cDF4, cEH30, CEH33, cEH38, and cEH2/cEH25, respectively). Northern blot analysis using the insert of 5kb EHOC-1 cDNA revealed three transcripts (5.3kb, 7.5kb and 8kb) on multiple adult tissues (heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas) . Fluorescence in-εitu hybridization was also performed on lymphocytes from a normal individual using the insert from the EHOC-1 5kb cDNA subclone as a probe. Discrete signals were seen on chromosome 21q22.3 confirming the loci. Tissue in-situ hybridization analysis of human and mouse EHOC-1 cDNA clones using human and mouse brain tissues revealed that EHOC-1 is expressed ubiquitously, but prominently in neuron cells. Both the dentate gyrus of hippocampus and the Purkinje cells of cerebellum in adult mice were tissues which showed fairly high signals. Because of marked loss of Purkinje cells in EPMl patients, the strong expression of EHOC-1 in Purkinje cells increase the candidacy of this gene as a cause of progressive myoclonus epilepsies. The complete sequence of the 5kb cDNA revealed an open reading frame of 3570bp (1190 amino acids;SEQ ID NO:2). The initiator ATG was located within a good Kozak consensus sequence Kozak, M. , J. Mol . Biol . 196:947-950 (1987); Kozak, M. , Nuc . Acid Res . 15:8125-8148 (1987) . A homology search of the amino acid sequence of this open reading frame (ORF) with genes registered in Genbank/EMBL showed that this gene product is homologous to multiple transmembrane proteins including three of sodium channel proteins (Figure 3) . A mouse fetal brain cDNA library (Stratagene, La Jolla, CA) was screened using an approximately 5kb human EHOC-1 cDNA clone (cDF9) as a probe. Two homologous cDNA clones were isolated and labeled cMOl (3.5 kb) and cM06 (3.0 kb) . The cM06 clone has been partially sequenced and is set forth in SEQ ID NO:6. The 229 base pair sequence set forth in SEQ ID NO:6 encodes a fragment of mouse EHOC-1 protein and is 75.4% homologous to human EHOC-1 cDNA at nucleotides 1913-2141 of SEQ ID N0:1. Some neurologic disorders in humans are known to result from mutations in sodium channels (Ptacek et al., Cell :1021-1027 (1991); Rojas et al., Nature 354:387-389 (1991); McClatchey et al., Ceil €>8_:769-774 (1992); Ptacek et al., Neuron 1:891-897 (1992)), calcium channels (Ptacek 11 et al.. Cell 72:863-868 (1994); Jurkut-Rott et al.. Hum . Mol . Genet . 3.:1415-1419 (1994)), and a potassium channel (Browne et al., Nature Genet . 1:136-140 (1994)). By using the BLAST computer program (Altschul, S.J., et al., J . Mol . Biol . 215:403-410 (1994)), one fibronectin domain (CxV YxC) was found at 356 < - > 401 a.a. The analysis also showed that the motif (Sxxx(I,L)E) occurs at 462, 670, 708, 716, 730, and 1078. This motif was further searched for in various protein databases and there were very few instances where it was present three or more times. This motif was present in, for example, rat cartilage specific proteoglycan core protein, myosin, drosophila sevenless (4 copies) , drosophila prospero (4 copies) , and drosophila serendipity (3 copies) . The latter three are mutants in development. The sevenless gene causes an eye defect, the prospero gene causes defects in axon pathfinding, and the serendipity gene results in defects in cellularization. It is reasonable, therefore, to assume that a defect in axonal routing may correlate with the phenotype of EHOC-l. The region beginning at 777 also shares some homology with multiple drug resistance genes and to the drosophila rutabaga gene. The rutabaga gene is associated with learning in drosophila . Accordingly, the present invention provides isolated nucleic acids, which encode a novel EHOC-l protein, wherein such nucleic acids are derived from human chromosome 21, specifically at the q23.2 locus, which is the site of mutation(s) that cause PME, HPE1, and APECED. The protein encoded by this gene has multiple transmembrane domains. An analysis of an approximately 5kb cDNA clone (cDF9) of EHOC-l revealed that the cDF9 consists of more than 23 exons. The term "nucleic acids" (also referred to as polynucleotides) encompasses RNA as well as single and double-stranded DNA and cDNA. As used herein, the phrase "isolated" means a nucleic acid that is in a form that does not occur in nature. One means of isolating a nucleic acid encoding an EHOC-l polypeptide is to probe a mammalian genomic library with a natural or artificially designed DNA probe using methods well known in the art. DNA probes derived from the EHOC-l gene are particularly useful for this purpose. DNA and cDNA molecules that encode EHOC-l polypeptides can be used to obtain complementary genomic DNA, cDNA or RNA from human, mammalian, or other animal sources, or to isolate related cDNA or genomic clones by the screening of cDNA or genomic libraries, by methods described in more detail below. Examples of nucleic acids are RNA, cDNA, or isolated genomic DNA encoding an EHOC-l polypeptide. Such nucleic acids may have coding sequences substantially the same as the coding sequence shown in SEQ ID NO:l or SEQ ID NO:6. This invention also encompasses nucleic acids which differ from the nucleic acids shown in SEQ ID NO:l or SEQ ID NO:6, but which have the same phenotype, i.e., those that encode all or a fragment of a protein that is substantially the same amino acid sequence set forth in SEQ ID NO:2. Phenotypically similar nucleic acids are also referred to as "functionally equivalent nucleic acids". As used herein, the phrase "functionally equivalent nucleic acids" encompasses nucleic acids characterized by slight and non-consequential sequence variations that will function in substantially the same manner to produce the same protein product(s) as the nucleic acids disclosed herein. In particular, functionally equivalent nucleic acids encode polypeptides that are the same as those disclosed herein or that have conservative amino acid variations. For example, conservative variations include substitution of a non-polar residue with another non-polar residue, or substitution of a charged residue with a similarly charged residue. These variations include those recognized by skilled artisans as those that do not substantially alter the tertiary structure of the protein. Further provided are nucleic acids encoding EHOC- 1 polypeptides that, by virtue of the degeneracy of the genetic code, do not necessarily hybridize to the invention nucleic acids under specified hybridization conditions. Preferred nucleic acids encoding the invention polypeptide are comprised of nucleotides that encode substantially the same amino acid sequence set forth in SEQ ID NO:2. Alternatively, preferred nucleic acids encoding the invention polypeptide(s) hybridize under high stringency conditions to substantially the entire sequence, or substantial portions (i.e., typically at least 15-30 nucleotides) of the nucleic acid sequence set forth in SEQ ID NO:l or SEQ ID NO:6. Stringency of hybridization, as used herein, refers to conditions under which polynucleotide hybrids are stable. As known to those of skill in the art, the stability of hybrids is a function of sodium ion concentration and temperature. (See, for example, Sambrook et al.. Molecular Cloning: A Laboratory Manual 2d Ed. (Cold Spring Harbor Laboratory, 1989; incorporated herein by reference) . Also provided are isolated peptides, polypeptides(s) and/or protein(s) encoded by the invention nucleic acids which are EHOC-l polypeptides. The EHOC-l polypeptide comprises a protein of approximately 1190 amino acids in length. The complete amino acid sequence encoding the human EHOC-l polypeptide is set forth in SEQ ID NO:2. In addition, SEQ ID NO:6 sets forth a 229 base pair mouse cDNA sequence that encodes a fragment of a mouse EHOC-l protein. As used herein, the term "isolated" means a protein molecule free of cellular components and/or contaminants normally associated with a native in vivo environment. Invention polypeptides and/or proteins include any natural occurring allelic variant, as well as recombinant forms thereof. The EHOC-l polypeptides can be isolated using various methods well known to a person of skill in the art. The methods available for the isolation and purification of invention proteins include, precipitation, gel filtration, ion-exchange, reverse-phase and affinity chromatography. Other well-known methods are described in Deutscher et al., Guide to Protein Purification: Methods in Enzvmolocrv Vol. 182, (Academic Press, 1990) , which is incorporated herein by reference. Alternatively, the isolated polypeptides of the present invention can be obtained using well-known recombinant methods as described, for example, in Sambrook et al., supra . , 1989) . An example of the means for preparing the invention polypeptide(ε) is to express nucleic acids encoding the EHOC-l in a suitable host cell, such as a bacterial cell, a yeast cell, an amphibian cell (i.e., oocyte) , or a mammalian cell, using methods well known in the art, and recovering the expressed polypeptide, again using well-known methods. Invention polypeptides can be isolated directly from cells that have been transformed with expression vectors, described below in more detail. The invention polypeptide, biologically active fragments, and functional equivalents thereof can also be produced by chemical synthesis. As used herein, "biologically active fragment" refers to any portion of the polypeptide represented by the amino acid sequence in SEQ ID NO:2 that can assemble into a cationic channel permeable to Ca + which is activated by acetylcholine. Synthetic polypeptides can be produced using Applied Biosyste s, Inc. Model 430A or 431A automatic peptide synthesizer (Foster City, CA) employing the chemistry provided by the manufacturer. As used herein, the phrase "EHOC-l" refers to naturally purified or recombinantly expressed/produced (i.e., isolated or substantially pure) proteins that contain highly hydrophobic regions which predict potential membrane spanning regions, having homologies to multiple transmembrane proteins, including sodium channel, calcium channel and potassium channel proteins including variants thereof encoded by mRNA generated by alternative splicing of a primary transcript, and further including fragments thereof which retain one or more of the aforementioned properties. As used herein, the phrase "functional polypeptide" means that binding of ligands, for example, cause transcriptional activation of EHOC-l proteins. More specifically, agonist activation of a "functional invention polypeptide" induces the protein to generate a signal. Modification of the invention nucleic acids, polypeptides or proteins with the following phrases: "recombinantly expressed/produced", "isolated", or "substantially pure", encompasses nucleic acids, peptides, polypeptides or proteins that have been produced in such form by the hand of man, and are thus separated from their native in vivo cellular environment. As a result of this human intervention, the recombinant nucleic acids, polypeptides and proteins of the invention are useful in ways that the corresponding naturally occurring molecules are not, such as identification of selective drugs or compounds. Sequences having "substantial sequence homology" are intended to refer to nucleotide sequences that share at least about 75%, preferably about 80%, yet more preferably about 90% identity with invention nucleic acids; and amino acid sequences that typically share at least about 75%, preferably about 85%, yet more preferably about 95% amino acid identity with invention polypeptides. It is recognized, however, that polypeptides or nucleic acids containing less than the above-described levels of homology arising as splice variants or that are modified by conservative amino acid substitutions, or by substitution of degenerate codons are also encompassed within the scope of the present invention. The present invention provides the isolated polynucleotide operatively linked to a promoter of RNA transcription, as well as other regulatory sequences. As used herein, the phrase "operatively linked" refers to the functional relationship of the polynucleotide with regulatory and effector sequences of nucleotides, such as promoters, enhancers, transcriptional and translational stop sites, and other signal sequences. For example, operative linkage of a polynucleotide to a promoter refers to the physical and functional relationship between the polynucleotide and the promoter such that transcription of DNA is initiated from the promoter by an RNA polymerase that specifically recognizes and binds to the promoter, and wherein the promoter directs the transcription of RNA from the polynucleotide. Promoter regions include specific sequences that are sufficient for RNA polymerase recognition, binding and transcription initiation. Additionally, promoter regions include sequences that modulate the recognition, binding and transcription initiation activity of RNA polymerase. Such sequences may be cis acting or may be responsive to trans acting factors. Depending upon the nature of the regulation, promoters may be constitutive or regulated. Examples of promoters are SP6, T4, T7, SV40 early promoter, cytomegalovirus (CMV) promoter, mouse mammary tumor virus (MMTV) steroid-inducible promoter, Moloney urine leukemia virus (MMLV) promoter, and the like. Vectors that contain both a promoter and a cloning site into which a polynucleotide can be operatively linked are well known in the art. Such vectors are capable of transcribing RNA in vitro or in vivo , and are commercially available from sources such as Stratagene (La Jolla, CA) and Pro ega Biotech (Madison, WI) . In order to optimize expression and/or in vitro transcription, it may be necessary to remove, add or alter 5* and/or 3' untranslated portions of the clones to eliminate extra, potential inappropriate alternative translation initiation codons or other sequences that may interfere with or reduce expression, either at the level of transcription or translation. Alternatively, consensus ribosome binding sites can be inserted immediately 5 ' of the start codon to enhance expression. (See, for example, Kozak, J. Biol. Chem. 266:19867 (1991)). Similarly, alternative codons, encoding the same amino acid, can be substituted for coding sequences of the EHOC-l polypeptide in order to enhance transcription (e.g., the codon preference of the host cell can be adopted, the presence of G-C rich domains can be reduced, and the like) . Also provided are vectors comprising invention nucleic acids. Examples of vectors are viruses, such as baculoviruses and retroviruses, bacteriophages, cosmids, plasmids and other recombination vehicles typically used in the art. Polynucleotides are inserted into vector genomes using methods well known in the art. For example, insert and vector DNA can be contacted, under suitable conditions, with a restriction enzyme to create complementary ends on each molecule that can pair with each other and be joined together with a ligase. Alternatively, synthetic nucleic acid linkers can be ligated to the termini of restricted polynucleotide. These synthetic linkers contain nucleic acid sequences that correspond to a particular restriction site in the vector DNA. Additionally, an oligonucleotide containing a termination codon and an appropriate restriction site can be ligated for insertion into a vector containing, for example, some or all of the following: a selectable marker gene, such as the neomycin gene for selection of stable or transient transfectants in mammalian cells; enhancer/promoter sequences from the immediate early gene of human CMV for high levels of transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; SV40 polyoma origins of replication and ColEl for proper episomal replication; versatile multiple cloning sites; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Other means are well known and available in the art. Further provided are vectors comprising nucleic acids encoding EHOC-l polypeptides, adapted for expression in a bacterial cell, a yeast cell, an amphibian cell (i.e., oocyte) , a mammalian cell and other animal cells. The vectors additionally comprise the regulatory elements necessary for expression of the nucleic acid in the bacterial, yeast, amphibian, mammalian or animal cells so located relative to the nucleic acid encoding EHOC-l polypeptide as to permit expression thereof. As used herein, "expression" refers to the process by which nucleic acids are transcribed into mRNA and translated into peptides, polypeptides, or proteins. If the nucleic acid is derived from genomic DNA, expression may include splicing of the mRNA, if an appropriate eucaryotic host is selected. Regulatory elements required for expression include promoter sequences to bind RNA polymerase and transcription initiation sequences for riboso e binding. For example, a bacterial expression vector includes a promoter such as the lac promoter and for transcription initiation the Shine-Dalgarno sequence and the start codon AUG (Sambrook et al. supra) . Similarly, a eucaryotic expression vector includes a heterologous or homologous promoter for RNA polymerase II, a downstream polyadenylation signal, the start codon AUG, and a termination codon for detachment of the ribosome. Such vectors can be obtained commercially or assembled by the sequences described in methods well known in the art, for example, the methods described above for constructing vectors in general. Expression vectors are useful to produce cells that express the invention polypeptide. The present invention provides transformed host cells that recombinantly express EHOC-l polypeptides. An example of a transformed host cell is a mammalian cell comprising a plasmid adapted for expression in a mammalian cell. The plasmid contains nucleic acid encoding an EHOC-l polypeptide and the regulatory elements necessary for expression of invention proteins. Various mammalian cells may be utilized as hosts, including, for example, mouse fibroblast cell NIH3T3, CHO cells, HeLa cells, Ltk- cells, etc. Expression plasmids such as those described supra can be used to transfect mammalian cells by methods well known in the art such as, for example, calcium phosphate precipitation, DEAE-dextran, electroporation, microinjection or lipofection. EHOC-l polypeptides expressed recombinantly on eucaryotic cell surfaces may contain at least one EHOC-l polypeptide, or may contain a mixture of peptides encoded by the host cell and/or subunits encoded by heterologous nucleic acids. The present invention provides nucleic acid probes comprising nucleotide sequences capable of specifically hybridizing with sequences included within nucleic acids encoding EHOC-l polypeptides, for example, a coding sequence included within the nucleotide sequence shown in SEQ ID N0:1 or SEQ ID NO:6. As used herein, a "probe" is a single-stranded DNA or RNA that has a sequence of nucleotides that includes at least 15 contiguous bases 15144 PCI7US95/14641 20 set forth in SEQ ID NO:l or SEQ ID NO:6. Preferred regions from which to construct probes include 5' and/or 3 1 coding sequences, sequences within the ORF, sequences predicted to encode transmembrane domains, sequences predicted to encode cytoplasmic loops, signal sequences, ligand binding sites, and the like. Full-length or fragments of cDNA clones can also be used as probes for the detection and isolation of related genes. When fragments are used as probes, preferably the cDNA sequences will be from the carboxyl end-encoding portion of the cDNA, and most preferably will include predicted transmembrane domain-encoding portions of the cDNA sequence. Transmembrane domain regions can be predicted based on hydropathy analysis of the deduced amino acid sequence using, for example, the method of Kyte and Doolittle, J. Mol. Biol. 157:105 (1982). In another embodiment, EHOC-l genomic DNA is contemplated for use as a diagnostic probe. For example, it has been found that a G (guanine) to A (adenosine) conversion occurs in the genomic intron sequence approximately 22 base pairs upstream of exon 7 in the EHOC- 1 gene. It is contemplated that this G to A genomic conversion is homozygous in PME patients. Thus, it is contemplated that such a mutation at a genomic nucleotide position corresponding to nucleotide 94 of SEQ ID NO:5 is associated with PME disease. The exon sequence at nucleotides 117-263 of SEQ ID NO:5 corresponds to nucleotides 1176-1322 of SEQ ID NO:l. Thus, any probe derived from SEQ ID NO:5 that can detect such a conversion at nucleotide 94 of SEQ ID NO:5 is contemplated for use herein. As used herein, the phrase "specifically hybridizing" encompasses the ability of a polynucleotide to recognize a sequence of nucleic acids that are complementary thereto and to form double-helical segments 15144 21 via hydrogen bonding between complementary base pairs. Nucleic acid probe technology is well known to those skilled in the art who will readily appreciate that such probes may vary greatly in length and may be labeled with a detectable agent, such as a radioisotope, a fluorescent dye, and the like, to facilitate detection of the probe. Invention probes are useful to detect the presence of nucleic acids encoding the EHOC-l polypeptide. For example, the probes can be used for in situ hybridizations in order to locate biological tissues in which the invention gene is expressed. Additionally, synthesized oligonucleotides complementary to the nucleic acids of a nucleotide sequence encoding EHOC-l polypeptide are useful as probes for detecting the invention genes, their associated mRNA, or for the isolation of related genes using homology screening of genomic or cDNA libraries, or by using amplification techniques well known to one of skill in the art. Also provided are antisense oligonucleotides having a sequence capable of binding specifically with any portion of an mRNA that encodes EHOC-l polypeptides so as to prevent translation of the mRNA. The antisense oligonucleotide may have a sequence capable of binding specifically with any portion of the sequence of the cDNA encoding EHOC-l polypeptides. As used herein, the phrase "binding specifically" encompasses the ability of a nucleic acid sequence to recognize a complementary nucleic acid sequence and to form double-helical segments therewith via the formation of hydrogen bonds between the complementary base pairs. An example of an antisense oligonucleotide is an antisense oligonucleotide comprising chemical analogs of nucleotides. Compositions comprising an amount of the antisense oligonucleotide, described above, effective to reduce expression of EHOC-l polypeptides by passing through a cell membrane and binding specifically with mRNA encoding EHOC-l polypeptides so as to prevent translation and an acceptable hydrophobic carrier capable of passing through a cell membrane are also provided herein. The acceptable hydrophobic carrier capable of passing through cell membranes may also comprise a structure which binds to a receptor specific for a selected cell type and is thereby taken up by cells of the selected cell type. The structure may be part of a protein known to bind to a cell-type specific receptor. Antisense oligonucleotide compositions are useful to inhibit translation of mRNA encoding invention polypeptides. Synthetic oligonucleotides, or other antisense chemical structures are designed to bind to mRNA encoding EHOC-l polypeptides and inhibit translation of mRNA and are useful as compositions to inhibit expression of EHOC-l associated genes in a tissue sample or in a subject. The present invention provides means to modulate levels of expression of EHOC-l polypeptides by employing synthetic antisense oligonucleotide compositions (hereinafter SAOC) which inhibit translation of mRNA encoding these polypeptides. Synthetic oligonucleotides, or other antisense chemical structures designed to recognize and selectively bind to mRNA, are constructed to be complementary to portions of the nucleotide sequences shown in SEQ ID NO:l or SEQ ID NO:6. The SAOC is designed to be stable in the blood stream for administration to a subject by injection, or in laboratory cell culture conditions. The SAOC is designed to be capable of passing through the cell membrane in order to enter the cytoplasm of the cell by virtue of physical and chemical properties of the SAOC which render it capable of passing through cell membranes, for example, by designing small, hydrophobic SAOC chemical structures, or by virtue of specific transport systems in the cell which recognize and transport the SAOC into the cell. In addition, the SAOC can be designed for administration only to certain selected cell populations by targeting the SAOC to be recognized by specific cellular uptake mechanisms which bind and take up the SAOC only within select cell populations. For example, the SAOC may be designed to bind to a receptor found only in a certain cell type, as discussed supra . The SAOC is also designed to recognize and selectively bind to target mRNA sequence, which may correspond to a sequence contained within the sequence shown in SEQ ID NO:l or SEQ ID NO:6. The SAOC is designed to inactivate target mRNA sequence by either binding thereto and inducing degradation of the mRNA by, for example, RNase I digestion, or inhibiting translation of mRNA target sequence by interfering with the binding of translation-regulating factors or ribosomes, or inclusion of other chemical structures, such as ribozyme sequences or reactive chemical groups which either degrade or chemically modify the target mRNA. SAOCs have been shown to be capable of such properties when directed against mRNA targets (see Cohen et al., TIPS, 10:435 (1989) and Weintraub, Sci. American. January (1990), pp.40; both incorporated herein by reference) . This invention also provides compositions containing an acceptable carrier and any of an isolated, purified EHOC-l polypeptide, an active fragment thereof, or a purified, mature protein and active fragments thereof, alone or in combination with each other. These polypeptides or proteins can be recombinantly derived, chemically synthesized or purified from native sources. As used herein, the term "acceptable carrier" encompasses any of the standard pharmaceutical carriers, such as phosphate buffered saline solution, water and emulsions such as an oil/water or water/oil emulsion, and various types of wetting agents. Further provided are antibodies having specific reactivity with EHOC-l polypeptides of the present invention. Active fragments of antibodies are encompassed within the definition of "antibody". Invention antibodies can be produced by methods known in the art using invention polypeptides, proteins or portions thereof as antigens. For example, polyclonal and monoclonal antibodies can be produced by methods well known in the art, as described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory 1988), which is incorporated herein by reference. Invention polypeptides can be used as immunogens in generating such antibodies. Alternatively, synthetic peptides can be prepared (using commercially available synthesizers) and used as immunogens. Amino acid sequences can be analyzed by methods well known in the art to determine whether they encode hydrophobic or hydrophilic domains of the corresponding polypeptide. Altered antibodies such as chimeric, humanized, CDR-grafted or bifunctional antibodies can also be produced by methods well known in the art. Such antibodies can also be produced by hybridoma, chemical synthesis or recombinant methods described, for example, in Sambrook et al., supra . , and Harlow and Lane, supra . Both anti-peptide and anti- fusion protein antibodies can be used, (see, for example, Bahouth et al., Trends Pharmacol. Sci. 12:338 (1991); Ausubel et al., Current Protocols in Molecular Biology (John Wiley and Sons, NY 1989) which are incorporated herein by reference) . Invention antibodies also can be used to isolate invention polypeptides. Additionally the antibodies are useful for detecting the presence of invention polypeptides, as well as analysis of chromosome localization, and structural as well as functional domains. Methods for detecting the presence of EHOC-l polypeptides on the surface of a cell comprise contacting the cell with an antibody that specifically binds to EHOC-l polypeptides, under conditions permitting binding of the antibody to the polypeptides, detecting the presence of the antibody bound to the cell, and thereby detecting the presence of invention polypeptides on the surface of the cell. With respect to the detection of such polypeptides, the antibodies can be used for in vitro diagnostic or in vivo imaging methods. Immunological procedures useful for in vitro detection of target EHOC-l polypeptides in a sample include immunoassays that employ a detectable antibody. Such immunoassays include, for example, ELISA, Pandex microfluorimetric assay, agglutination assays, flow cytometry, serum diagnostic assays and immunohistochemical staining procedures which are well known in the art. An antibody can be made detectable by various means well known in the art. For example, a detectable marker can be directly or indirectly attached to the antibody. Useful markers include, for example, radionucleotides, enzymes, fluorogens, chromogens and che iluminescent labels. Further, invention antibodies can be used to modulate the activity of the EHOC-l polypeptide in living animals, in humans, or in biological tissues or fluids isolated therefrom. Accordingly, compositions comprising a carrier and an amount of an antibody having specificity for EHOC-l polypeptides effective to block binding of naturally occurring ligands to invention polypeptides. A monoclonal antibody directed to an epitope of EHOC-l polypeptide molecules present on the surface of a cell and having an amino acid sequence substantially the same as an amino acid sequence for a cell surface epitope of an EHOC-l polypeptide shown in SEQ ID NO:2, can be useful for this purpose. The present invention further provides transgenic non-human mammals that are capable of expressing nucleic acids encoding EHOC-l polypeptides. Also provided are transgenic non-human mammals capable of expressing nucleic acids encoding EHOC-l polypeptides so mutated as to be incapable of normal activity, i.e., do not express native EHOC-l. The present invention also provides transgenic non-human mammals having a genome comprising antisense nucleic acids complementary to nucleic acids encoding EHOC-l polypeptides so placed as to be transcribed into antisense mRNA complementary to mRNA encoding EHOC-l polypeptides, which hybridizes thereto and, thereby, reduces the translation thereof. The nucleic acid may additionally comprise an inducible promoter and/or tissue specific regulatory elements, so that expression can be induced, or restricted to specific cell types. Examples of nucleic acids are DNA or cDNA having a coding sequence substantially the same as the coding sequence shown in SEQ ID NO:l or SEQ ID NO:6. An example of a non-human transgenic mammal is a transgenic mouse. Examples of tissue specificity-determining elements are the metallothionein promoter and the L7 promoter. Animal model systems which elucidate the physiological and behavioral roles of EHOC-l polypeptides are produced by creating transgenic animals in which the expression of the EHOC-l polypeptide is altered using a variety of techniques. Examples of such techniques include the insertion of normal or mutant versions of nucleic acids encoding an EHOC-l polypeptide by microinjection, retroviral infection or other means well known to those skilled in the art, into appropriate fertilized embryos to produce a transgenic animal. (See, for example, Hogan et al. , Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory, 1986) . Another technique, homologous recombination of mutant or normal versions of these genes with the native gene locus in transgenic animals, may be used to alter the regulation of expression or the structure of EHOC-l polypeptides (see, Capecchi et al.. Science 244:1288 (1989); Zim er et al.. Nature 338:150 (1989) ; which are incorporated herein by reference) . Homologous recombination techniques are well known in the art. Homologous recombination replaces the native (endogenous) gene with a recombinant or mutated gene to produce an animal that cannot express native (endogenous) protein but can express, for example, a mutated protein which results in altered expression of EHOC-l polypeptides. In contrast to homologous recombination, microinjection adds genes to the host genome, without removing host genes. Microinjection can produce a transgenic animal that is capable of expressing both endogenous and exogenous EHOC-l protein. Inducible promoters can be linked to the coding region of nucleic acids to provide a means to regulate expression of the transgene. Tissue specific regulatory elements can be linked to the coding region to permit tissue-specific expression of the transgene. Transgenic animal model systems are useful for in vivo screening of compounds for identification of specific ligands, i.e., agonists and antagonists, which activate or inhibit protein responses. Invention nucleic acids, oligonucleotides (including antisense) , vectors containing same, transformed host cells, polypeptides and combinations thereof, as well as antibodies of the present invention, can be used to screen compounds in vitro to determine whether a compound functions as a potential agonist or antagonist to invention polypeptides. These in vitro screening assays provide information regarding the function and activity of invention polypeptides, which can lead to the identification and design of compounds that are capable of specific interaction with one or more types of polypeptides, peptides or proteins. In accordance with still another embodiment of the present invention, there is provided a method for identifying compounds which bind to EHOC-l polypeptides. The invention proteins may be employed in a competitive binding assay. Such an assay can accommodate the rapid screening of a large number of compounds to determine which compounds, if any, are capable of binding to EHOC-l proteins. Subsequently, more detailed assays can be carried out with those compounds found to bind, to further determine whether such compounds act as modulators, agonists or antagonists of invention proteins. In another embodiment of the invention, there is provided a bioassay for identifying compounds which modulate the activity of invention polypeptides. According to this method, invention polypeptides are contacted with an "unknown" or test substance (in the presence of a reporter gene construct when antagonist activity is tested) , the activity of the polypeptide is monitored subsequent to the contact with the "unknown" or test substance, and those substances which cause the reporter gene construct to be expressed are identified as functional ligands for EHOC-l polypeptides. In accordance with another embodiment of the present invention, transformed host cells that recombinantly express invention polypeptides can be contacted with a test compound, and the modulating effect(s) thereof can then be evaluated by comparing the EHOC-1-mediated response (via reporter gene expression) in the presence and absence of test compound, or by comparing the response of test cells or control cells (i.e., cells that do not express EHOC-l polypeptides) , to the presence of the compound. As used herein, a compound or a signal that "modulates the activity" of invention polypeptides refers to a compound or a signal that alters the activity of EHOC- 1 polypeptides so that the activity of the invention polypeptide is different in the presence of the compound or signal than in the absence of the compound or signal. In particular, such compounds or signals include agonists and antagonists. An agonist encompasses a compound or a signal that activates EHOC-l protein expression. Alternatively, an antagonist includes a compound or signal that interferes with EHOC-l protein expression. Typically, the effect of an antagonist is observed as a blocking of agonist-induced protein activation. Antagonists include competitive and non-competitive antagonists. A competitive antagonist (or competitive blocker) interacts with or near the site specific for agonist binding. A non-competitive antagonist or blocker inactivates the function of the polypeptide by interacting with a site other than the agonist interaction site. As understood by those of skill in the art, assay methods for identifying compounds that modulate EHOC-l activity generally require comparison to a control. One type of a "control" is a cell or culture that is treated substantially the same as the test cell or test culture exposed to the compound, with the distinction that the "control" cell or culture is not exposed to the compound. For example, in methods that use voltage clamp electrophysiological procedures, the same cell can be tested in the presence or absence of compound, by merely changing the external solution bathing the cell. Another type of "control" cell or culture may be a cell or culture that is identical to the transfected cells, with the exception that the "control" cell or culture do not express native proteins. Accordingly, the response of the transfected cell to compound is compared to the response (or lack thereof) of the "control" cell or culture to the same compound under the same reaction conditions. In yet another embodiment of the present invention, the activation of EHOC-l polypeptides can be modulated by contacting the polypeptides with an effective amount of at least one compound identified by the above- described bioassays. The invention will now be described in greater detail with reference to the following non-limiting examples. EXAMPLE 1 Construction of BAC Contig BAC library construction of total human genomic DNA was performed as described in Shizuya et al., Proc. Natl . Acad. Sci . USA .89:8794-8797 (1992). BAC clones were screened by PCR using STSs (PFKL, D21S25, D21S154, CD18) . The loci of these BAC clones were confirmed by fluorescence in-εitu hybridization. Insert size of BAC clones was measured by running pulsed-field gel electrophoresis after digesting DNA with Notl. EXAMPLE 2 Direct cDNA Selection Direct selection procedures were similar to those described in Morgan et al., Nucleic Acid Res . 20:5173-5179 (1992) with some modifications. Total RNA was isolated from 14 week trisomy 21 fetal brain using TRI region TM (Molecular Research Center, Inc.). Poly (A) * RNA was isolated using Poly (A) Quick mRNA isolation kit (STRATAGENE) . Double stranded cDNA was synthesized using Superscript TM Choice System (GIBCO BRL) from 5μg trisomy 21 fetal brain poly (A) * RNA using lμg oligo (dT) 15 or O.lμg random hexamer. The entire synthesis reaction was purified by Gene Clean II kit (BI0101, Inc.) and then kinased. Sau3AI linker was attached to the cDNA which was subsequently digested with Sau3AI. The reaction was purified using Gene Clean. Mbol linker was attached to the cDNA and the reaction purified by Gene Clean (Morgan et al.. Nucleic Acid Res . .20:5173-5179 (1992)). The synthesized product was amplified by PCR using one strand Of Mbol linker (5'CCTGATGCTCGAGTGAATTC3•) (SEQ ID NO:3) as a primer. PCR cycling conditions were 40 cycles of 94°C/15 seconds, 60°C/23 seconds, 72°C/2 minutes in a lOOμl of lx PCR buffer (Promega) , 3mM MgCl 2 , 5.0 units of Tag polymerase (Promega) , 2μM primer and 0.2mM dNTPs. Five BAC DNAs (total 2.5μg) were prepared using QIAGEN plasmid kit and were biotinylated using Nick Translation Kit and biotin-16-dUTP (Boehringer Mannehei ) . 3μg of heat denatured PCR amplified cDNA was annealed with 3μg of heat denatured COTl DNA (BRL) in lOOμl hybridization buffer (750mM NaCl, 50mM NaPO (pH7.2) , 5mM EDTA, 5x Denhardt's, 0.05% SDS and 50% formamide) at 42°C for two hours. After prehybridization, 1.2μg of heat denatured biotinylated BAC DNA was added and incubated at 42°C for 16 hours. cDNA-BAC DNA hybrids were precipitated with EtOH and dissolved in 60μl of lOmM Tris-HCl (pH 8.0), ImM EDTA. After addition of 40μl 5M NaCl, the DNA was incubated with magnetic beads (Dynabeads M-280, Dynal) at 25°C for 1 hour with gentle rotating to allow attachment of the DNA to the magnetic beads. The beads were then washed twice by pipetting in 400μl of 2x SSC, setting in magnet holder (MPC-E TM , Dynal) for 30 seconds and removing the supernatant. Four additional washes were performed in 0.2x SSC at 68°C for 10 minutes each with transfer of the beads to new tubes at each wash. cDNAs were eluted in lOOμl of distilled water for 10 minutes at 80°C with occasional mixing. The eluted cDNAs were amplified by PCR as described above. After twice repeating the selection procedure using magnetic beads, amplified cDNAs were digested with EcoRI and subcloned into pBluescript II. EXAMPLE 3 Southern blot analysis Gel electrophoresis of DNA was carried out on 0.8% agarose gels in lx TBE. Transfer of nucleic acids to Nybond N+ nylon membrane (Amersham) was performed according to the manufacturer's instruction. Probes were labelled using RadPri e Labeling System (BRL) . Hybridization was carried out at 42°C for 16 hours in 50% forma ide, 5x SSPE, 5x Denhardt's 0.1% SDS, lOOμg/ml denatured salmon sperm DNA. The filters were washed once in lx SSC, 0.1% SDS at room temperature for 20 minutes, and twice in O.lx SSC, 0.1% SDS for 20 minutes at 65°C. The blots were exposed onto X-ray film (Kodak, X-OMAT-AR) . EXAMPLE 4 cDNA Library Screening A trisomy 21 fetal brain cDNA library was constructed using ZAP-cDNA synthesis kit (STRATAGENE) which generates a unidirectional cDNA library. Briefly, double- stranded cDNA was synthesized from 5μg trisomy 21 fetal brain poly(A) * RNA using a hybrid oligo(dT)-Xhol linker primer with 5-methyl dCTP. An EcoRI linker was attached to the cDNA which was subsequently digested with EcoRI and Xhol, and then cloned into UNI-ZAP XR vector. The library * was packaged using Gigapack II Gold packaging extract. The titer of the original library was 1.1 x 10 p.f.u./package. The library was amplified once. A blue-white color assay indicated that 99% of the clones had inserts. The average size of the inserts was 1.9 kb, as calculated from 14 clones. Screening of the trisomy 21 fetal brain cDNA library was performed using selected cDNA fragments. Phages were plated to an average density of 1 x 10 per 175 cm 2 plate. Plaque lifts of 20 plates (2 x 10 phages) were made using duplicated nylon membranes (Hybond-N+; Amersham) . Hybridized membranes were washed to final stringency of 0.2x SSC, O.lx SDS at 65°C. The filters were exposed overnight onto X-ray film. Phages were subcloned into the plasmid vector pBluescript II SK(-) by M13- mediated excision for further analysis. EXAMPLE 5 Northern Blot Analysis cDNA inserts were excised from the vector by digestion with Xhol and EcoRI. After labeling using the random priming method, the fragments were used a probes for Northern hybridization using Multiple Tissue Northern Blot (Clontech) . EXAMPLE 6 Metaphase Preparation Chromosomes were prepared using a BrdU block, (Zabel et al. in Proc . Natl . Acad . Sci . USA 80:6932-6936 (1983)) with some modification. Briefly, human peripheral lymphocytes were grown for 72 hours at 37°C in RPMI 1640 (GIBCO BRL, Gaithersburg, MD) supplemented with L-glutamine (2mM) , 15% fetal calf serum, penicillin (100 IU/ml) , streptomycin (0.05mg/ml) and 0.02% phytohemagglutinin. The cells were blocked in S-phase by adding 5-bromo- deoxyuridine (0.8mg/ml) for 16 hours. Cells were washed once with HBSS (Hanks Balanced Salt Solution) (GIBCO BRL, Gaithersburg, MD) to remove the synchronizing agent and were released by incubating for five to six more hours in medium supplemented with 2.5μg/ml thy idine. Cultures were harvested by the addition of O.lμg/ml colcemid for 10 minutes followed by 0.075 M KCl hypotonic solution for 15 minutes at 37°C prior to fixation with a 3:1 mixture of methanol and acetic acid, for 1-5 minutes. EXAMPLE 7 High Quality Chromosome Preparation To obtain high quality chromosome preparations, the metaphase spreads were prepared by letting one drop of suspension fall onto alcohol-cleaned slides, evenly and flat, completely free of cytoplasm. The slides were then placed above a container filled with heated water for 20-60 seconds depending on the ambient humidity. The optimum conditions were determined by first checking several slides under a phase contrast microscope. For best results, the slides should be aged at room temperature for at least 2-3 weeks prior to in situ hybridization. Slides are best stored at -70°C after aging. The day before denaturation, slides are removed from -70°C and kept overnight at 4°C. Slides are then left at room temperature for 1-2 hours before baking them at 55-60°C for 2 hours, followed by denaturation in 70% formamide (at 66-70°C) . For best results, fresh slides are denatured at 66°C. EXAMPLE 8 Probe Fragment Size for Mapping Small DNA Probes The fragment size of a probe labeled by nick translation is preferably around 100-200 bp and the concentration of probe DNA in the hybridization mixture is preferably in the range of 20-40 ng/μl (i.e., 200-400 ng/slide) . This concentration can increase non-specific binding, but generally will produce good signal/noise ratio. For reducing non-specific background hybridization of repeat sequences, a small amount of CotI DNA can be used with cDNA probes (i.e., l-3μg of CotI with 200-400 ng of probe DNA) . EXAMPLE 9 Biotinylation of Probes DNA probes were labeled with biotin-14-dATP by nick translation (GIBCO BRL) . Unincorporated nucleotides were separated by chromatography (Sephadex G-50) . The mean fragment size of all DNA probes was set at around 200 bp, with a range of 100-600 bp, established by nondenaturing agarose gels. EXAMPLE 10 In situ Hybridization To obtain specific hybridization of single-copy sequences with small DNA probes, the FISH method described by Lichter et al. in Science 247:64-69 (1990) was modified. RNase treatment is unnecessary for slides made more than one year prior to their use. When necessary, RNase treatment consisted of lOOμg/ml RNase, placed on slides for 20 minutes at 37°C, followed by dehydration through a cold ethanol series of 70%, 90% and 100%. The denaturation of slides was performed at 67-70°C in 70% formamide/2x SSC (0.15 M NaCl, 0.015 M Sodium Citrate) for 1-2 minutes (fresh chromosome preparations require a shorter time) . The hybridization buffer (lOμl) containing the biotinylated probes together with 3μg Cot 1 DNA and 7μg sonicated salmon sperm DNA, (the ratio of probe: Cot 1 DNA is 1:15-30; total DNA concentration at lμg/μl) was placed on each slide. A coverslip was then applied and sealed with rubber cement, followed by overnight incubation in a humidified chamber at 37°C. EXAMPLE 11 Detection of Probe Signals by I munofluorescence After overnight hybridization, the slides were washed at 44°C in 50% formamide (v/v) /2X SSC (4 x 5 minutes) and then at 50-60°C in 2X SSC (3 x 5 minutes) . The rinsing time was shorter (3 x 2 minutes) for small probes (insert size <1.5 Kb). To increase the intensity of the fluorescence signal produced by smaller probes, it is necessary to reduce the stringency of post-hybridization washes (2X SSC, at 45-50°C) . Slides were then incubated for 10-20 minutes at 37°C with lOOμl of 4X SSC containing 3% bovine serum albumin (BSA) and 0.1% Tween 20 for blocking. The buffer was then replaced with FITC-Avidin (0.5μg/ml in 4X SSC/1% BSA/0.1% Tween 20), for 30 minutes at 37°C. After three 5 minutes washes at 45 °C in 2X SSC/0.1% Tween 20, the hybridization signal was amplified by the addition of a biotinylated goat anti-avidin antibody layer (5μg/ml) for 30 minutes at 37°C. The slides were preincubated in 5% goat serum/2X SSC/3% BSA/0.1% Tween 20 for 10-20 minutes prior to the amplification step to reduce i munological non-specific binding. After washing, a second layer of FITC-avidin was added to the slides as previously described. To increase the intensity of the hybridization signal, a second round of amplification can be applied as necessary. The slides were then washed in 2X SSC/0.1% Tween 20 three times at 45°C and briefly left to drain. EXAMPLE 12 Chromosome R-Banding To view the chromosome bands and fluorescence signals simultaneously, the dyes chromomycin A3 and dista ycin A were used as counterstain (Schweizer in Chromoεoma 51:307-324 (1976); Schweizer in Hum . Genet 57:1-14 (1981); and Magenis et al. in Hum . Genet 69:300-303 (1985)). Immediately following the last detection, the slides were rinsed briefly in Mcllavane's buffer (pH 8.5- 9.0) Schweizer et al., supra , (diluted 1:1 with distilled water) prior to staining. Subsequently, lOOμl of chromomycin A3 (0.5mg/ml in 1/2 Mcllavane's buffer pH 8.5- 9.0) was placed on slides for 10 minutes - 1 hour at room temperature in the dark. The staining time required depends on the freshness of the slides; fresher slides need longer staining times, whereas aged slides may need only 10 minutes. After a first round of staining, the slides were rinsed for 1 minute in 1/2 Mcllavane's buffer and the excess fluid was shaken off. This was followed by a second round of staining by placing 50μl of O.lmg/ml distamycin A on the slide, incubating for 1-2 minutes at room temperature, followed by rinsing, as above. If using DNA probes having small inserts (e.g., expected to produce faint signals) , at this point, the slides can be mounted with a thin layer of anti-fade solution containing p-phenylenediamine in phosphate buffer (Johnson et al. in J. Immunol. Methods 4_3:349-350 (1981)). Maximum banding resolution and minimal fading is obtained by two rounds of chromomycin A3 and distamycin A staining. In addition, the best banding results are obtained by staining the slides immediately following the detection step, but no later than 2-3 days. Slides are best viewed through the microscope immediately after staining, but no later than 3 days for small probes (1.0 Kb - 2.5 Kb) or 1-2 weeks for larger probes (>2.5-3 Kb). Only a light counterstain is needed before examining the slide under a fluorescence microscope. In fact, an advantage of this banding technique is that the counterstaining step can be repeated at any point after hybridization to make the chromosomes brighter, if necessary. Further, after image capture, using computerized systems, the banding pattern may be selectively bleached to more clearly view the FITC hybridization signal. EXAMPLE 13 Microscopy and Photography The slides were viewed with a Zeiss Axiophot 100 or Axiovert 135 fluorescence microscope (Zeiss, Inc., Thornwood, NY) . The FITC and chromomycin A3 are both excited by using a 400-490nm band pass exciter, 460nm dichroic, and 470nm barrier (Zeiss filter set #05) . The hybridized segments appear as bright green-blue or bright yellow-green spots, while the rest of the chromosome bands appear dim green. Kodak Technical pan (ASA100) film was used for black and white photographs. Color images were captured by a cooled-CCD camera (Photometries CH 250, Photometries Ltd., Tuscon, AZ) using the BDS (Biological Detection Systems, Pittsburgh, PA) imaging software. While the invention has been described in detail with reference to certain preferred embodiments thereof, it will be understood that modifications and variations are within the spirit and scope of that which is described and claimed. 39 SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: KORENBERG, JULIE R. YAMAKAWA, KAZUHIRO (ii) TITLE OF INVENTION: A NOVEL CHROMOSOME 21 GENE MARKER, COMPOSITIONS AND METHODS USING SAME (iii) NUMBER OF SEQUENCES: 6 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: PRETTY, SCHROEDER, BRUEGGEMANN & CLARK (B) STREET: 444 SOUTH FLOWER STREET, SUITE 2000 (C) CITY: LOS ANGELES (D) STATE: CA (E) COUNTRY: USA (F) ZIP: 90071 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 08/337,690 (B) FILING DATE: 09-NOV-1994 (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: REITER, STEPHEN E. (B) REGISTRATION NUMBER: 31,192 (C) REFERENCE/DOCKET NUMBER: FP41 90079 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 619-546-1995 (B) TELEFAX: 619-546-9392 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5107 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 138..3710 (D) OTHER INFORMATION: /product= "EHOC-l" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GCGGCGCAAC CGGCTCCGGA GCTGCCTGGC GCGGCCGGGC GGGCGGCGCC GCTCAGGCTC 60 GGGCTCCGGC TGGGCCCGGC GCGGCCTCGG GGCTGCCCAT GGGGCGCGGG GGGCCGGGCC 12 GGTGACGCCG GACGCCC ATG GAC GCC TCT GAG GAG CCG CTG CCG CCG GTG 170 Met Asp Ala Ser Glu Glu Pro Leu Pro Pro Val 1 5 10 ATC TAC ACC ATG GAG AAC AAG CCC ATC GTC ACC TGT GCT GGA GAT CAG 218 lie Tyr Thr Met Glu Asn Lys Pro lie Val Thr Cys Ala Gly Asp Gin 15 20 25 AAT TTA TTT ACC TCT GTT TAT CCA ACG CTC TCT CAG CAG CTT CCA AGA 26 Asn Leu Phe Thr Ser Val Tyr Pro Thr Leu Ser Gin Gin Leu Pro Arg 30 35 40 GAA CCA ATG GAA TGG AGA AGG TCC TAT GGC CGG.GCT CCG AAG ATG ATT 314 Glu Pro Met Glu Trp Arg Arg Ser Tyr Gly Arg Ala Pro Lys Met lie 45 50 55 CAC CTA GAG TCT AAC TTT GTT CAA TTC AAA GAG GAG CTG CTG CCC AAA 362 His Leu Glu Ser Asn Phe Val Gin Phe Lys Glu Glu Leu Leu Pro Lys 60 65 70 75 GAA GGA AAC AAA GCT CTG CTC ACG TTT CCC TTC CTC CAT ATT TAC TGG 410 Glu Gly Asn Lys Ala Leu Leu Thr Phe Pro Phe Leu His lie Tyr Trp 80 85 90 ACA GAG TGC TGT GAT ACC GAA GTG TAT AAA GCT ACA GTA AAA GAT GAC 458 Thr Glu Cys Cys Asp Thr Glu Val Tyr Lys Ala Thr Val Lys Asp Asp 95 100 105 CTC ACC AAG TGG CAG AAT GTT CTG AAG GCT CAT AGC TCT GTG GAC TGG 506 Leu Thr Lys Trp Gin Asn Val Leu Lys Ala His Ser Ser Val Asp Trp 110 115 120 TTA ATA GTG ATA GTT GAA AAT GAT GCC AAG AAA AAA AAC AAA ACC AAC 554 Leu lie Val lie Val Glu Asn Asp Ala Lys Lys Lys Asn Lys Thr Asn 125 130 135 ATC CTT CCC CGA ACC TCT ATT GTG GAC AAA ATA AGA AAT GAT TTT TGT 602 lie Leu Pro Arg Thr Ser lie Val Asp Lys lie Arg Asn Asp Phe Cys 140 145 150 155 AAT AAA CAG AGT GAC AGG TGT GTT GTG CTC TCC GAC CCC TTG AAG GAC 650 Asn Lys Gin Ser Asp Arg Cys Val Val Leu Ser Asp Pro Leu Lys Asp 160 165 170 TCT TCT CGA ACT CAG GAA TCC TGG AAT GCC TTC CTG ACC AAA CTC AGG 698 Ser Ser Arg Thr Gin Glu Ser Trp Asn Ala Phe Leu Thr Lys Leu Arg 175 180 185 ACA TTG CTT CTT ATG TCT TTT ACC AAA AAC CTA GGC AAG TTT GAG GAT 746 Thr Leu Leu Leu Met Ser Phe Thr Lys Asn Leu Gly Lys Phe Glu Asp 190 195 200 GAC ATG AGA ACC TTG AGG GAG AAG AGG ACT GAG CCA GGC TGG AGC TTT 794 Asp Met Arg Thr Leu Arg Glu Lys Arg Thr Glu Pro Gly Trp Ser Phe 205 210 215 TGT GAA TAT TTC ATG GTT CAG GAG GAG CTT GCC TTT GTT TTC GAG ATG 842 Cys Glu Tyr Phe Met Val Gin Glu Glu Leu Ala Phe Val Phe Glu Met 220 225 230 235 CTG CAG CAG TTC GAG GAC GCC CTG GTG CAG TAC GAC GAA CTG GAC GCC 890 Leu Gin Gin Phe Glu Asp Ala Leu Val Gin Tyr Asp Glu Leu Asp Ala 240 245 250 CTC TTC TCT CAG TAT GTG GTC AAC TTC GGG GCC GGG GAT GGT GCC AAC 938 Leu Phe Ser Gin Tyr Val Val Asn Phe Gly Ala Gly Asp Gly Ala Asn 255 260 265 TGG CTG ACT TTT TTC TGC CAG CCA GTG AAG AGC TGG AAC GGA TTG ATC 986 Trp Leu Thr Phe Phe Cys Gin Pro Val Lys Ser Trp Asn Gly Leu lie 270 275 280 CTC CGA AAA CCC ATA GAT ATG GAG AAG CGG GAA TCG ATC CAG AGG CGA 1034 Leu Arg Lys Pro lie Asp Met Glu Lys Arg Glu Ser lie Gin Arg Arg 285 290 295 GAA GCC ACC CTG TTA GAT CTG CGC AGT TAC CTG TTC TCT CGC CAG TGC 1082 Glu Ala Thr Leu Leu Asp Leu Arg Ser Tyr Leu Phe Ser Arg Gin Cys 300 305 310 315 ACC TTG CTG CTC TTC CTG CAG AGG CCG TGG GAG GTG GCC CAG CGC GCC 1130 Thr Leu Leu Leu Phe Leu Gin Arg Pro Trp Glu Val Ala Gin Arg Ala 320 325 330 CTA GAG CTG CTG CAC AAC TGC GTG CAG GAA CTG AAG CTC TTA GAA GTC 1178 Leu Glu Leu Leu His Asn Cys Val Gin Glu Leu Lys Leu Leu Glu Val 335 340 345 TCT GTC CCA CCT GGT GCT CTG GAC TGC TGG GTG TTT CTG AGC TGT CTG 1226 Ser Val Pro Pro Gly Ala Leu Asp Cys Trp Val Phe Leu Ser Cys Leu 350 355 360 GAG GTG TTG CAG AGG ATA GAA GGC TGC TGT GAC CGG GCA CAG ATC GAC 1274 Glu Val Leu Gin Arg lie Glu Gly Cys Cys Asp Arg Ala Gin lie Asp 365 370 375 TCA AAC ATT GCC CAC ACT GTG GGG CTA TGG AGC TAT GCC ACA GAA AAG 1322 Ser Asn lie Ala His Thr Val Gly Leu Trp Ser Tyr Ala Thr Glu Lys 380 385 390 395 TTA AAG TCC TTG GGC TAT CTA TGT GGA CTT GTG TCA GAG AAA GGA CCT 1370 Leu Lys Ser Leu Gly Tyr Leu Cys Gly Leu Val Ser Glu Lys Gly Pro 400 405 410 AAC TCA GAA GAT CTC AAC AGG ACA GTT GAC CTT TTG GCA GGT TTG GGA 1418 Asn Ser Glu Asp Leu Asn Arg Thr Val Asp Leu Leu Ala Gly Leu Gly 415 420 425 GCT GAG CGA CCA GAA ACA GCC AAC ACA GCT CAG AGT CCT TAT AAG AAA 1466 Ala Glu Arg Pro Glu Thr Ala Asn Thr Ala Gin Ser Pro Tyr Lys Lys 430 435 440 CTG AAA GAA GCA TTA TCG TCA GTG GAA GCT TTT GAA AAA CAC TAC TTA 1514 Leu Lys Glu Ala Leu Ser Ser Val Glu Ala Phe Glu Lys His Tyr Leu 445 450 455 GAT TTG TCC CAT GCC ACC ATT GAA ATG TAT ACA AGC ATT GGG AGG ATT 1562 Asp Leu Ser His Ala Thr He Glu Met Tyr Thr Ser He Gly Arg He 460 465 470 475 CGA TCT GCT AAG TTT GTT GGA AAA GAT CTG GCA GAG TTT TAC ATG AGG 1610 Arg Ser Ala Lys Phe Val Gly Lys Asp Leu Ala Glu Phe Tyr Met Arg 480 485 490 AAA AAG GCT CCA CAA AAG GCA GAA ATC TAT CTT CAA GGA GCA CTG AAA 1658 Lys Lys Ala Pro Gin Lys Ala Glu He Tyr Leu Gin Gly Ala Leu Lys 495 500 505 AAC TAC CTG GCT GAG GGC TGG GCA CTC CCC ATC ACA CAC ACA AGG AAG 1706 Asn Tyr Leu Ala Glu Gly Trp Ala Leu Pro He Thr His Thr Arg Lys 510 515 520 CAG CTG GCC GAA TGT CAA AAG CAC CTT GGA CAA ATT GAA AAC TAC CTG 1754 Gin Leu Ala Glu Cys Gin Lys His Leu Gly Gin He Glu Asn Tyr Leu 525 530 535 CAG ACC AGC AGC CTC TTA GCC AGT GAC CAC CAC CTC ACT GAA GAG GAG 1802 Gin Thr Ser Ser Leu Leu Ala Ser Asp His His Leu Thr Glu Glu Glu 540 545 550 555 CGC AAG CAC TTC TGC CAG GAG ATA CTT GAC TTT GCC AGC CAG CCG TCA 1850 Arg Lys His Phe Cys Gin Glu He Leu Asp Phe Ala Ser Gin Pro Ser 560 565 570 GAC AGC CCA GGT CAT AAG ATA GTG CTA CCC ATG CAT TCC TTT GCA CAA 1898 Asp Ser Pro Gly His Lys He Val Leu Pro Met His Ser Phe Ala Gin 575 580 585 CTG CGA GAT CTC CAT TTT GAT CCC TCC AAT GCC GTG GTC CAC GTG GGC 1946 Leu Arg Asp Leu His Phe Asp Pro Ser Asn Ala Val Val His Val Gly 590 595 600 GGC GTT TTG TGC GTT GAG ATA ACC ATG TAC AGC CAG ATG CCT GTG CCT 1994 Gly Val Leu Cys Val Glu He Thr Met Tyr Ser Gin Met Pro Val Pro 605 610 615 GTT CAC GTG GAG CAG ATT GTG GTC AAT GTC CAC TTC AGC ATT GAG AAA 2042 Val His Val Glu Gin He Val Val Asn Val His Phe Ser He Glu Lys 620 625 630 635 AAC AGC TAC CGG AAG ACT GCG GAG TGG CTT ACC AAG CAC AAG ACG TCC 2090 Asn Ser Tyr Arg Lys Thr Ala Glu Trp Leu Thr Lys His Lys Thr Ser 640 645 650 AAT GGG ATC ATT AAC TTT CCA CCC GAG ACC GCA CCT TTC CCT GTA TCC 2138 Asn Gly He He Asn Phe Pro Pro Glu Thr Ala Pro Phe Pro Val Ser 655 660 665 CAA AAC AGT TTG CCC GCG CTG GAG TTG TAT GAA ATG TTT GAG AGA AGC 2186 Gin Asn Ser Leu Pro Ala Leu Glu Leu Tyr Glu Met Phe Glu Arg Ser 670 675 680 CCA TCT GAT AAC TCC TTG AAC ACG ACT GGG ATT ATC TGC AGA AAC GTC 2234 Pro Ser Asp Asn Ser Leu Asn Thr Thr Gly He He Cys Arg Asn Val 685 690 695 CAC ATG CTC CTG AGA AGG CAG GAG AGC AGC TCC TCT CTA GAG ATG CCC 2282 His Met Leu Leu Arg Arg Gin Glu Ser Ser Ser Ser Leu Glu Met Pro 700 705 710 715 TCA GGG GTG GCT CTG GAG GAG GGT GCC CAC GTG CTG AGG TGC AGC CAC 2330 Ser Gly Val Ala Leu Glu Glu Gly Ala His Val Leu Arg Cys Ser His 720 725 730 GTG ACC CTG GAA CCA GGG GCC AAC CAG ATA ACA TTC AGG ACT CAG GCC 2378 Val Thr Leu Glu Pro Gly Ala Asn Gin He Thr Phe Arg Thr Gin Ala 735 740 745 AAG GAA CCT GGA ACG TAT ACA CTC AGG CAG CTG TGC GCC TCG GTG GGC 2426 Lys Glu Pro Gly Thr Tyr Thr Leu Arg Gin Leu Cys Ala Ser Val Gly 750 755 760 TCC GTG TGG TTC GTC CTC CCT CAC ATC TAC CCC ATT GTG CAG TAC GAC 2474 Ser Val Trp Phe Val Leu Pro His He Tyr Pro He Val Gin Tyr Asp 765 770 775 GTG TAC TCA CAG GAG CCC CAG CTG CAC GTG GAG CCG CTG GCT GAT AGC 2522 Val Tyr Ser Gin Glu Pro Gin Leu His Val Glu Pro Leu Ala Asp Ser 780 785 790 795 CTT CTG GCA GGC ATT CCT CAG AGA GTC AAG TTC ACT GTC ACT ACC GGC 2570 Leu Leu Ala Gly He Pro Gin Arg Val Lys Phe Thr Val Thr Thr Gly 800 805 810 CAT GAT ACG ATA AAG AAT GGA GAC AGC CTG CAG CTT AGC AAT GCC GAA 2618 His Asp Thr He Lys Asn Gly Asp Ser Leu Gin Leu Ser Asn Ala Glu 815 820 825 GCC ATG CTC ATC CTG TGC CAG GCG GAG AGC AGG GCT GTG GTC TAC TCC 2666 Ala Met Leu He Leu Cys Gin Ala Glu Ser Arg Ala Val Val Tyr Ser 830 835 840 AAC ACG AGA GAA CAG TCT TCT GAG GCC GCG CTC CGG ATT CAG TCC TCC 2714 Asn Thr Arg Glu Gin Ser Ser Glu Ala Ala Leu Arg He Gin Ser Ser 845 850 855 GAC AAG GTC ACG AGC ATC AGT CTG CCT GTT GCG CCT GCG TAC CAC GTG 2762 Asp Lys Val Thr Ser He Ser Leu Pro Val Ala Pro Ala Tyr His Val 860 865 870 875 ATC GAA TTT GAA CTG GAA GTT CTC TCT TTA CCT TCA GCC CCA GCA CTC 2810 He Glu Phe Glu Leu Glu Val Leu Ser Leu Pro Ser Ala Pro Ala Leu 880 885 890 GGA GGG GAG AGT GAC ATG CTG GGG ATG GCA GAG CCC CAC AGG AAG CAT 2858 Gly Gly Glu Ser Asp Met Leu Gly Met Ala Glu Pro His Arg Lys His 895 900 905 AAG GAC AAA CAG AGA ACT GGC CGC TGC ATG GTT ACC ACA GAC CAC AAA 2906 Lys Asp Lys Gin Arg Thr Gly Arg Cys Met Val Thr Thr Asp His Lys 910 915 920 GTG TCG ATT GAC TGC CCG TGG TCC ATC TAC TCC ACA GTC ATC GCA CTG 2954 Val Ser He Asp Cys Pro Trp Ser He Tyr Ser Thr Val He Ala Leu 925 930 935 ACC TTC AGC GTA CCC TTC AGG ACC ACA CAC AGC CTC CTG TCC TCA GGA 3002 Thr Phe Ser Val Pro Phe Arg Thr Thr His Ser Leu Leu Ser Ser Gly 940 945 950 955 ACA CGG AAA TAT GTT CAA GTT TGT GTC CAG AAT TTG TCA GAA CTT GAC 3050 Thr Arg Lys Tyr Val Gin Val Cys Val Gin Asn Leu Ser Glu Leu Asp 960 965 970 TTT CAG CTG TCA GAT AGT TAT CTT GTA GAT ACC GGT GAT AGT ACC GAC 3098 Phe Gin Leu Ser Asp Ser Tyr Leu Val Asp Thr Gly Asp Ser Thr Asp 975 980 985 CTG CAA CTA GTA CCA CTG AAC ACG CAG TCC CAG CAG CCC ATC TAC AGC 3146 Leu Gin Leu Val Pro Leu Asn Thr Gin Ser Gin Gin Pro He Tyr Ser 990 995 1000 AAG CAG TCG GTG TTC TTC GTC TGG GAA CTC AAG TGG ACA GAA GAG CCT 3194 Lys Gin Ser Val Phe Phe Val Trp Glu Leu Lys Trp Thr Glu Glu Pro 1005 1010 1015 CCC CCT TCT CTG CAT TGC CGG TTC TCT GTT GGA TTT TCC CCA GCT TCT 3242 Pro Pro Ser Leu His Cys Arg Phe Ser Val Gly Phe Ser Pro Ala Ser 1020 1025 1030 1035 GAG GAA CAG CTG TCT ATC TCC TTA AAG CCG TAT ACT TAT GAA TTT AAA 3290 Glu Glu Gin Leu Ser He Ser Leu Lys Pro Tyr Thr Tyr Glu Phe Lys 1040 1045 1050 GTG GAA AAT TTT TTT ACA TTA TAC AAC GTG AAG GCT GAG ATC TTT CCC 3338 Val Glu Asn Phe Phe Thr Leu Tyr Asn Val Lys Ala Glu He Phe Pro 1055 1060 1065 CCT TCG GGA ATG GAG TAT TGC AGA ACA GGC TCC CTC TGC TCC CTG GAG 3386 Pro Ser Gly Met Glu Tyr Cys Arg Thr Gly Ser Leu Cys Ser Leu Glu 1070 1075 1080 GTT TTG ATC ACG AGG CTC TCA GAC CTC TTG GAG GTG GAT AAA GAT GAA 3434 Val Leu He Thr Arg Leu Ser Asp Leu Leu Glu Val Asp Lys Asp Glu 1085 1090 1095 GCA CTG ACT GAA TCT GAT GAG CAT TTT TCG ACA AAG CTT ATG TAT GAA 3482 Ala Leu Thr Glu Ser Asp Glu His Phe Ser Thr Lys Leu Met Tyr Glu 1100 1105 1110 1115 GTT GTC GAC AAC AGT AGC AAC TGG GCA GTG TGT GGG AAA AGC TGC GGT 3530 Val Val Asp Asn Ser Ser Asn Trp Ala Val Cys Gly Lys Ser Cys Gly 1120 1125 1130 GTC ATC TCC ATG CCA GTG GCT GCT CGG GCC ACT CAC AGG GTC CAC ATG 3578 Val He Ser Met Pro Val Ala Ala Arg Ala Thr His Arg Val His Met 1135 1140 1145 GAA GTG ATG CCG CTC TTC GCC GGG TAT CTC CCC CTG CCC GAC GTC AGG 3626 Glu Val Met Pro Leu Phe Ala Gly Tyr Leu Pro Leu Pro Asp Val Arg 1150 1155 1160 CTG TTC AAG TAC CTC CCC CAT CAT TCT GCA CAC TCC TCC CAA CTG GAC 3674 Leu Phe Lys Tyr Leu Pro His His Ser Ala His Ser Ser Gin Leu Asp 1165 1170 1175 GCT GAC AGC TGG ATA GAA AAC GCA GCC TGT CAG TAGACAAGCA CGGGGACGAC 3727 Ala Asp Ser Trp He Glu Asn Ala Ala Cys Gin 1180 1185 1190 CAGCCGGACA GCAGCAGCCT CAAGAGCAGG GGCAGCGTGC ATTCGGCCTG CAGCAGCGAG 3787 CACAAAGGCC TACCCATGCC CCGGCTGCAG GCACTGCCGG CCGGCCAGGT CTTCAACTCC 3847 AGCTCGGGCA CACAAGTCCT GGTCATCCCC AGCCAAGATG ACCACGTCCT GGAAGTCAGT 3907 GTAACATGAC AACGCCAGGG TGAACACACG CCACTTCCCA GCTAGGAGTG CACTTTATGG 3967 GACTGTGACT GGACTCTTCC GTTCTGGCTC CAGCCAGACC TTCAGTGGTC CTGCCTGGCC 4027 GTGGGGACAT CAGAGAGTGT CATCACGCAG CTGGCCAGCT GAGTTCTGTT GTTGTTTTCA 4087 TGCCGCCTGT GATCTCAGAT TCCTGCTTTT CTCACCCCGT CCCCATGCTG GTGTCCGACG 4147 CCGCTTACTC AGAGCCCTGG CCTCCCTCCC CCTACCTCAC ACGCTGCTCA TGAAAGTTTC 4207 CACCCACGCT GTCTCCACGG AACAGCCTCC GTCTGCTGGC TCTTCGTGGA AGGCCATTTG 4267 TCTTTCAGGT AGACACTCAG CAGCCCTCAC GGTCTTAGTG ACGTGTGTGC CTTTCTGGTC 4327 ACACAGCTGC CCAGTTTCCT GATCGGGGTG GATTTGTGTC CCCTAAGGGG TAAAACAGCC 4387 GTTTACCGCA GATCCTCTCA TACACCCTTC TAGGGGAGGC GGGTGGGGGA GGGAGGGATC 4447 ATAACCCCTT CTGTGCCTTG GGATGCCGGA GCTGGGGGAC CTGGAGGCCC ATCAGCCGGA 4507 GCCACGTGAA AGGTACTGAA GAAAGCTGAG ACCCGGCTGT GAGGAGCGCC TCAGCGGTGA 4567 GGTGGTTTAG GGATAAATGT TTCTGGAACC CTGTGGTCCC CCATAATGTT GATAGATATC 4627 ATATGCACTG GGAGTTAAAT ATATTTAATT TAATGATCAT TATATATGTG GGGGTTAATA 4687 TGTTGTTTTT CTGTCCCTTT AAAGTCTTTA CATGTAATTG TAGCTGTATA ATCGTTATTT 4747 TTCTTTTGCA TCTTAAGTCT TAGAAATTAA GATATTCCAT CGTGAGGATG AGAGAGGTCC 4807 TCAGTGTGTT TTTGGTCTGG TTGTAGGGAA GGACTCAAGT CCTGGAATGT CCTCCACTGG 4867 TCTACTGAGT TGCAGTCACA CTGTTCCAAT GGATTATTTG CTTTCGGTTG TAAATTTAAT 4927 TGTACATATG GTTGATTTAT TATTTTTAAA AATACAGACT AACTGATGTA ATGTTTATGT 4987 ATAAGTTGCA CCAAAAATCA AGGACAAAAA TAAGTGTGTT TGTTTTTACA GGTGTGAAAG 5047 TCACAGCTTG TAAATAAGTG TTGTATGTAT TAAACCTTTT CCAGTTCTCC AAAGCGATGT 5107 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1190 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Asp Ala Ser Glu Glu Pro Leu Pro Pro Val He Tyr Thr Met Glu 1 5 10 15 Asn Lys Pro He Val Thr Cys Ala Gly Asp Gin Asn Leu Phe Thr Ser 20 25 30 Val Tyr Pro Thr Leu Ser Gin Gin Leu Pro Arg Glu Pro Met Glu Trp 35 40 45 Arg Arg Ser Tyr Gly Arg Ala Pro Lys Met He His Leu Glu Ser Asn 50 55 60 Phe Val Gin Phe Lys Glu Glu Leu Leu Pro Lys Glu Gly Asn Lys Ala 65 70 75 80 Leu Leu Thr Phe Pro Phe Leu His He Tyr Trp Thr Glu Cys Cys Asp 85 90 95 Thr Glu Val Tyr Lys Ala Thr Val Lys Asp Asp Leu Thr Lys Trp Gin 100 105 110 Asn Val Leu Lys Ala His Ser Ser Val Asp Trp Leu He Val He Val 115 120 125 Glu Asn Asp Ala Lys Lys Lys Asn Lys Thr Asn He Leu Pro Arg Thr 130 135 140 Ser He Val Asp Lys He Arg Asn Asp Phe Cys Asn Lys Gin Ser Asp 145 150 155 160 Arg Cys Val Val Leu Ser Asp Pro Leu Lys Asp Ser Ser Arg Thr Gin 165 170 175 Glu Ser Trp Asn Ala Phe Leu Thr Lys Leu Arg Thr Leu Leu Leu Met 180 185 190 Ser Phe Thr Lys Asn Leu Gly Lys Phe Glu Asp Asp Met Arg Thr Leu 195 200 205 Arg Glu Lys Arg Thr Glu Pro Gly Trp Ser Phe Cys Glu Tyr Phe Met 210 215 220 Val Gin Glu Glu Leu Ala Phe Val Phe Glu Met Leu Gin Gin Phe Glu 225 230 235 240 Asp Ala Leu Val Gin Tyr Asp Glu Leu Asp Ala Leu Phe Ser Gin Tyr 245 250 255 Val Val Asn Phe Gly Ala Gly Asp Gly Ala Asn Trp Leu Thr Phe Phe 260 265 270 Cys Gin Pro Val Lys Ser Trp Asn Gly Leu He Leu Arg Lys Pro He 275 280 285 Asp Met Glu Lys Arg Glu Ser He Gin Arg Arg Glu Ala Thr Leu Leu 290 295 300 Asp Leu Arg Ser Tyr Leu Phe Ser Arg Gin Cys Thr Leu Leu Leu Phe 305 310 315 320 Leu Gin Arg Pro Trp Glu Val Ala Gin Arg Ala Leu Glu Leu Leu His 325 330 335 Asn Cys Val Gin Glu Leu Lys Leu Leu Glu Val Ser Val Pro Pro Gly 340 345 350 Ala Leu Asp Cys Trp Val Phe Leu Ser Cys Leu Glu Val Leu Gin Arg 355 360 365 He Glu Gly Cys Cys Asp Arg Ala Gin He Asp Ser Asn He Ala His 370 375 380 Thr Val Gly Leu Trp Ser Tyr Ala Thr Glu Lys Leu Lys Ser Leu Gly 385 390 395 400 Tyr Leu Cys Gly Leu Val Ser Glu Lys Gly Pro Asn Ser Glu Asp Leu 405 410 415 Asn Arg Thr Val Asp Leu Leu Ala Gly Leu Gly Ala Glu Arg Pro Glu 420 425 430 Thr Ala Asn Thr Ala Gin Ser Pro Tyr Lys Lys Leu Lys Glu Ala Leu 435 440 445 Ser Ser Val Glu Ala Phe Glu Lys His Tyr Leu Asp Leu Ser His Ala 450 455 460 Thr He Glu Met Tyr Thr Ser He Gly Arg He Arg Ser Ala Lys Phe 465 470 475 480 Val Gly Lys Asp Leu Ala Glu Phe Tyr Met Arg Lys Lys Ala Pro Gin 485 490 495 Lys Ala Glu He Tyr Leu Gin Gly Ala Leu Lys Asn Tyr Leu Ala Glu 500 505 510 Gly Trp Ala Leu Pro He Thr His Thr Arg Lys Gin Leu Ala Glu Cys 515 520 525 Gin Lys His Leu Gly Gin He Glu Asn Tyr Leu Gin Thr Ser Ser Leu 530 535 540 Leu Ala Ser Asp His His Leu Thr Glu Glu Glu Arg Lys His Phe Cys 545 550 555 560 Gin Glu He Leu Asp Phe Ala Ser Gin Pro Ser Asp Ser Pro Gly His 565 570 575 Lys He Val Leu Pro Met His Ser Phe Ala Gin Leu Arg Asp Leu His 580 585 590 Phe Asp Pro Ser Asn Ala Val Val His Val Gly Gly Val Leu Cys Val 595 600 605 Glu He Thr Met Tyr Ser Gin Met Pro Val Pro Val His Val Glu Gin 610 615 620 He Val Val Asn Val His Phe Ser He Glu Lys Asn Ser Tyr Arg Lys 625 630 635 640 Thr Ala Glu Trp Leu Thr Lys His Lys Thr Ser Asn Gly He He Asn 645 650 655 Phe Pro Pro Glu Thr Ala Pro Phe Pro Val Ser Gin Asn Ser Leu Pro 660 665 670 Ala Leu Glu Leu Tyr Glu Met Phe Glu Arg Ser Pro Ser Asp Asn Ser 675 680 685 Leu Asn Thr Thr Gly He He Cys Arg Asn Val His Met Leu Leu Arg 690 695 700 Arg Gin Glu Ser Ser Ser Ser Leu Glu Met Pro Ser Gly Val Ala Leu 705 710 715 720 Glu Glu Gly Ala His Val Leu Arg Cys Ser His Val Thr Leu Glu Pro 725 730 735 Gly Ala Asn Gin He Thr Phe Arg Thr Gin Ala Lys Glu Pro Gly Thr 740 745 750 Tyr Thr Leu Arg Gin Leu Cys Ala Ser Val Gly Ser Val Trp Phe Val 755 760 765 Leu Pro His He Tyr Pro He Val Gin Tyr Asp Val Tyr Ser Gin Glu 770 775 780 Pro Gin Leu His Val Glu Pro Leu Ala Asp Ser Leu Leu Ala Gly He 785 790 795 800 Pro Gin Arg Val Lys Phe Thr Val Thr Thr Gly His Asp Thr He Lys 805 810 815 Asn Gly Asp Ser Leu Gin Leu Ser Asn Ala Glu Ala Met Leu He Leu 820 825 830 Cys Gin Ala Glu Ser Arg Ala Val Val Tyr Ser Asn Thr Arg Glu Gin 835 840 845 Ser Ser Glu Ala Ala Leu Arg He Gin Ser Ser Asp Lys Val Thr Ser 850 855 860 He Ser Leu Pro Val Ala Pro Ala Tyr His Val He Glu Phe Glu Leu 865 870 875 880 Glu Val Leu Ser Leu Pro Ser Ala Pro Ala Leu Gly Gly Glu Ser Asp 885 890 895 Met Leu Gly Met Ala Glu Pro His Arg Lys His Lys Asp Lys Gin Arg 900 905 910 Thr Gly Arg Cys Met Val Thr Thr Asp His Lys Val Ser He Asp Cys 915 920 925 Pro Trp Ser He Tyr Ser Thr Val He Ala Leu Thr Phe Ser Val Pro 930 935 940 Phe Arg Thr Thr His Ser Leu Leu Ser Ser Gly Thr Arg Lys Tyr Val 945 950 955 960 Gin Val Cys Val Gin Asn Leu Ser Glu Leu Asp Phe Gin Leu Ser Asp 965 970 975 Ser Tyr Leu Val Asp Thr Gly Asp Ser Thr Asp Leu Gin Leu Val Pro 980 985 990 Leu Asn Thr Gin Ser Gin Gin Pro He Tyr Ser Lys Gin Ser Val Phe 995 1000 1005 Phe Val Trp Glu Leu Lys Trp Thr Glu Glu Pro Pro Pro Ser Leu His 1010 1015 1020 Cys Arg Phe Ser Val Gly Phe Ser Pro Ala Ser Glu Glu Gin Leu Ser 1025 1030 1035 1040 He Ser Leu Lys Pro Tyr Thr Tyr Glu Phe Lys Val Glu Asn Phe Phe 1045 1050 1055 Thr Leu Tyr Asn Val Lys Ala Glu He Phe Pro Pro Ser Gly Met Glu 1060 1065 1070 Tyr Cys Arg Thr Gly Ser Leu Cys Ser Leu Glu Val Leu He Thr Arg 1075 1080 1085 Leu Ser Asp Leu Leu Glu Val Asp Lys Asp Glu Ala Leu Thr Glu Ser 1090 1095 HOO Asp Glu His Phe Ser Thr Lys Leu Met Tyr Glu Val Val Asp Asn Ser 1105 1110 1115 1120 Ser Asn Trp Ala Val Cys Gly Lys Ser Cys Gly Val He Ser Met Pro 1125 1130 1135 Val Ala Ala Arg Ala Thr His Arg Val His Met Glu Val Met Pro Leu 1140 1145 1150 Phe Ala Gly Tyr Leu Pro Leu Pro Asp Val Arg Leu Phe Lys Tyr Leu 1155 1160 1165 Pro His His Ser Ala His Ser Ser Gin Leu Asp Ala Asp Ser Trp He 1170 1175 1180 Glu Asn Ala Ala Cys Gin 1185 1190 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: CCTGATGCTC GAGTGAATTC 20 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1801 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vii) IMMEDIATE SOURCE: (B) CLONE: CEH25 (EHOC-l 3' Untranslated cDNA) (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 10 (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 310 (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 1149 (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 1155 (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 1798 (ix) FEATURE: (A) NAME/KEY: polyA_site (B) LOCATION: 1801 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: ATTTTTGTAC ACTTGAAATA GAGTACTCTT AATTTACTGG GCAAATGTGC TTGGAATTGA 60 ACTTGACAAG ATTAGCTCAA GCAGATAGAG TCGGGTCCAG CAGTGGGTGG CCCTCGTGTG 120 AATCCCCGTG GATGTGCAAG TTGTGGAGAG AAGGAGCACC GGGTTCCTGC CCAGCACTGT 180 GCTTGCGGGA GGCGGTGGGG CATGGGAGGA AGGAGGCACA GACCGGGGAA ATATGACAGC 240 CGTCATTTCC AGTATTCTCT GTGTTGTCTT TTAGCTCATT CAATAAATAA AGGTGGTGTG 300 ATTTTTTTTC CTCCTGTCTT TTTCATTTGT AGAAACTGGA GACGTGTAAA GAGATAAATA 360 ATTGGGTAAT TAAACTTTCC AGAAATTTAT CTTCCTCATG TGCAGTTAAC AAACTTGGTC 420 AAACTAGTTA GCAAATTAGA ACTTCAGAAT CTAATGATAG TTTAGGGTTT CTAANATAAG 480 GTTTNTTATT GTAAANATTG ACGATTGCCC TGCATTTCTA CCAAGTCCTG TGAATAAAGA 540 GATGGGAGTT TGATTCCGTC AGAAGAGACT GTAATCCGTG TCGTCAGCCT GGGAGCCTTC 600 CCCNGNGTAA TGTAGCTTTC TCTCTTACCT TCTGGAAGAG GGAATGTTTC ATTTATTACT 660 GTTGATTTCT TGTATCTGGT TCTACTCCCA GGATGAAATT ATCCAACTAC ATATATATTT 720 AGAGGAAGAA AGTGAAGGGG AAATTTAAAA TGTTTACGGC GCTTAATTGC CTGGAAATGA 780 AATGAAATCA AATTTATCAG TTTTTTTCCC CCTAATTACC CAAAAGATCT TTTGCAAACT 840 ATGTTACATG AATGCTTCTG CCTCTTTAAG ACAAAGAAGA ATGTCACCCA AAATTGTCAT 900 TTTTTTCTTA ATGTTCATCA TAAAAGTCCT AAAAAGGGTA ACCGTAATTG GATGTTTATT 960 GTTTTTATCT AAAGTAAGGT GTATGTGTTT GAGACAAGCT GGTTTTGTTG ATAAAGAGAT 1020 GTTAAATAAT TGTGAAGCCA GATATGCAAT GTGTATCTAA AAAGCAAGGA ATTTGCAGCC 1080 GTTTTACAAA TATCTGTGGA ACATGTAAAT ACTGTCAAAT GGAAAATAAA ATAAGTTATA 1140 ATTTTTGTGA ATTTCATGGG ATGTCCTATG ATTGGAAAAA TTATAACTCT TCTGATTCTA 1200 ATGTGGAAAT TGTATTTAAT CTGAAAATGA CTTTACCTAC AACAGTTCCA TTGTCAGCAC 1260 AGCCTAGGAG GGTCAGATTC CGTATTAATT GCTCTTAGTG GAGATGCCAG ATATCCCATA 1320 CAGAATTAGC AGAGAAAATA CAGACAGGCT TCTATTCAAA TTTTCTTTAG TGCTTAAAAT 1380 TAAGTTTTAA AATGAAATCA GACACTGCAG GTTTGTATAT AAAATGAAAA GCTATACTAC 1440 TTTTTATAAA AGGGCAAACT GGGCTGATGT AAATGTTTTA CTTTCAACTG TGTTCTTTAA 1500 AATAAATCCT ACCTGGTTTT TAAATTTTAT TTTTCATGAA AATGCTCCTT TCTCTACATT 1560 TATTCATCCT ATATACATCA GGCTGTAAGA CCCCCCCCAG TCATCATTAA TACAATGTGT 1620 TGGGATTCTG TGACTGGAAA AGGTGACAAG TTGGTGACTT TGACACTGCA GGTATTCCAT 1680 TTTCATGGTT TACTATGAAA AGTCATTTTT CATATTATGT AATATATTGT TAGATTAAAA 1740 CCATTGTATT AAGACTTTAA AATGTAAGCA TTGTAATTCT GAAAATACAC ATTTTAAGAA 1800 G 1801 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 469 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE: (A) ORGANISM: HUMAN (viii) POSITION IN GENOME: (A) CHROMOSOME/SEGMENT: 21q22.3 (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 117..263 (D) OTHER INFORMATION: /product= "EXON 7 OF EHOC-l cDNA" (ix) FEATURE: (A) NAME/KEY: intron (B) LOCATION: 1..116 (ix) FEATURE: (A) NAME/KEY: exon (B) LOCATION: 117..263 ( ix ) FEATURE : (A) NAME/KEY: intron (B) LOCATION: 264..469 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: AGATGAGGTG GCTGCTTGAA GGTGAGGTTT GCACGTTCAA GCTAGAGCAT GTGGGTTTGT 60 TCAAGCTAGA GCTGTCCCGG AGCACCCCTC ACACGTTCGC ATTTGCACCC CCACAGGTCT 120 CTGTCCCACC TGGTGCTCTG GACTGCTGGG TGTTTCTGAG CTGTCTGGAG GTGTTGCAGA 180 GGATAGAAGG CTGCTGTGAC CGGGCACAGA TCGACTCAAA CATTGCCCAC ACTGTGGGGC 240 TATGGAGCTA TGCCACAGAA AAGGTGCCTA CCTGCCCAAG TGTGGAATGC TCACGTTGTC 300 TCTGCGGCCA TGCCTGGGTG GCGGAGGAAG TCTGCTGTTT GGAGGAGAGG TGTTGCTCAT 360 TTAGATCACG ATGCATCCAC TTTAGTGGCC CTAGAAGGTG TCTGGGTGCA GCCAAAGAAG 420 TCATAGTTCC CTACCACATG TCGATGTAGT CAGCAGACAG CAAGCACTT 469 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 229 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vii) IMMEDIATE SOURCE: (A) LIBRARY: MOUSE FETAL BRAIN cDNA (B) CLONE: CM06 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..229 (D) OTHER INFORMATION: /product= "MOUSE EHOC-l" /note= "This mouse cDNA region has 75.4% homology to human cDNA corresponding to nucleotides 1913-2141 of SEQ ID NO:l." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: TTTTGACCCT CCCAATGCTG TGGTCCATGC GGGTGGTGTN CTGACTGTGG AGATAACGGT 60 ATACAGCCAG ATGCCCATCC CTGTCCACNT NGACCAGATT NCTGTCAATG TNCACTTNAG 120 CATCGAAAAA AACAACTACC GGAAGACAGC CGNNNNGCTG ACCAAGCACA AGACTTCCAA 180 TGGAATCATC ACCTTNNCAG CTGANNNCTC ACTGTTNCCT GCATCTCAG 229

Claims: What is claimed is: 1. Isolated nucleic acid encoding a human EHOC- 1 polypeptide. 2. Isolated nucleic acid according to claim 1, wherein said nucleic acid comprises DNA. 3. DNA according to claim 2, wherein said DNA is a cDNA. 4. DNA according to claim 2, wherein said DNA encodes the amino acid sequence set forth in SEQ ID NO:2. 5. DNA according to claim 2, wherein said DNA hybridizes under high stringency conditions to substantially the entire coding sequence (nucleotides 138- 3710) set forth in SEQ ID N0:1 or set forth in SEQ ID NO:6. 6. DNA according to claim 2, wherein said DNA has substantially the same nucleotide sequence as the nucleotide sequence set forth in SEQ ID NO:l or SEQ ID NO:6. 7. A vector comprising DNA according to claim 8. A host cell containing a vector according to claim 7 , wherein said cell is a procaryotic cell or a eucaryotic cell. 9. A host cell according to claim 8, wherein said cell expresses a functional EHOC-l protein. 10. A nucleic acid probe comprising at least 15 nucleotides capable of specifically hybridizing with a sequence of nucleic acids of the nucleotide sequence set forth in SEQ ID N0:1 or SEQ ID NO:6. 11. A nucleic acid probe according to claim 10, wherein said probe is labeled with a detectable marker. 12. A kit for detecting mutations and aneuploidies in chromosome 21 at locus q22.3 comprising a plurality of probes, wherein each probe comprises a nucleic acid sequence having at least 15 bp of contiguous nucleotides capable of specifically hybridizing with a sequence of nucleic acids of the nucleotide sequence set forth in SEQ ID N0:1 or SEQ ID NO:6, and wherein each individual probe corresponds to a specific locus on chromosome 2lq22.3. 13. Isolated mRNA complementary to DNA according to claim 2. 14. An oligonucleotide composition comprising chemical analogues of the nucleic acid of claim 2 operatively linked to a promoter of RNA transcription. 15. An antisense oligonucleotide capable of specifically binding to and modulating the translation of mRNA according to claim 13. 16. Isolated EHOC-l polypeptide and functional equivalents thereof. 17. Isolated EHOC-l polypeptide according to claim 16, wherein said polypeptide has substantially the same amino acid sequence as that set forth in SEQ ID NO:2. 18. Isolated EHOC-l polypeptide according to claim 16, wherein said polypeptide has the same amino acid sequence as that set forth in SEQ ID NO:2. 19. Isolated EHOC-l polypeptide according to claim 16, wherein said polypeptide is encoded by a nucleotide sequence that is substantially the same nucleotide sequence as that set forth in SEQ ID N0:1 or SEQ ID NO:6. 20. Isolated EHOC-l polypeptide according to claim 16, wherein said polypeptide is encoded by the nucleotide sequence set forth in SEQ ID NO:l or SEQ ID NO:6. 21. An EHOC-l polypeptide expressed recombinantly in a host cell. 22. An EHOC-l polypeptide according to claim 21, wherein said polypeptide is encoded by a nucleotide sequence that is substantially the same as the nucleotide sequence set forth in SEQ ID NO:l or SEQ ID NO:6. 23. An EHOC-l polypeptide according to claim 21, wherein said polypeptide is encoded by the nucleotide sequence set forth in SEQ ID NO:l or SEQ ID NO:6. 24. An antibody that specifically binds to a determinant on a human EHOC-l protein or active fragment thereof. 25. An antibody according to claim 24, wherein said antibody is a monoclonal antibody. 26. An antibody according to claim 24, wherein said antibody is a polyclonal antibody. 27. A composition comprising an amount of the antisense oligonucleotide according to claim 13 effective to modulate expression of a human EHOC-l polypeptide and an acceptable hydrophobic carrier capable of passing through a cell membrane. 28. A composition according to claim 27, wherein the oligonucleotide is coupled to a substance which inactivates mRNA. 29. A composition according to claim 28, wherein said substance is a ribozyme. 30. A composition comprising an amount of an antibody according to claim 24 effective to block binding of naturally occurring ligands to the human EHOC-l receptor and an acceptable carrier. 31. A transgenic nonhuman mammal expressing DNA encoding a human EHOC-l polypeptide. 32. A transgenic nonhuman mammal according to claim 31, wherein said DNA encoding said polypeptide has been mutated as to be incapable of normal polypeptide activity, and wherein the polypeptide so expressed is not native EHOC-l polypeptide. 33. A transgenic nonhuman mammal, the genome of which comprising antisense DNA complementary to DNA encoding a human EHOC-l polypeptide, wherein said antisense DNA is transcribed into antisense mRNA complementary to mRNA encoding a human EHOC-l polypeptide. 34. A transgenic nonhuman mammal according to claim 31, wherein said DNA is operatively linked to an inducible promoter. 35. A transgenic nonhuman mammal according to claim 31, wherein said DNA is operatively linked to tissue specific regulatory elements. 36. A transgenic nonhuman mammal according to claim 31, wherein the transgenic nonhuman mammal is a mouse. 37. A method for identifying nucleic acids encoding a human EHOC-l protein, said method comprising: contacting a sample containing nucleic acids with a probe according to claim 11, wherein said contacting is effected under high stringency hybridization conditions, and identifying compounds which hybridize thereto. 38. A method for identifying compound(s) which bind to a human EHOC-l polypeptide, said method comprising contacting cells according to claim 9 with said compound(s) and identifying compounds which bind thereto. 39. A method for detecting the presence of a human EHOC-l polypeptide on a cell surface, said method comprising contacting a test cell with an antibody according to claim 24, detecting the presence of an antibody-receptor complex, and therefor detecting the presence of a human EHOC-l polypeptide on the cell surface. 40. A method for diagnosing a predisposition to a disorder associated with the expression of a specific human EHOC-l polypeptide allele, said method comprising: contacting a sample containing nucleic acids with a plurality of probes, wherein each probe comprises a nucleic acid sequence having at least 15 bp of contiguous nucleotides capable of specifically hybridizing with a sequence of nucleic acids of the nucleotide sequence set forth in SEQ ID NO:l or SEQ ID NO:6, and wherein each individual probe corresponds to a specific locus on chromosome 21q22.3. 41. A method according to claim 40, wherein said disorder is selected from progressive myoclonus epilepsy, holoprosencephaly, or autoimmune polyglandular disease. 42. A method for deterring the onset of symptoms associated with particular disorder comprising administering a composition which modulates expression of gene. 43. A method for introducing changes at human chromosome locus 21q22.3 comprising transforming a sample of cells obtained from a subject having progressive myoclonus epilepsy with the nucleic acid according to claim 1 along with a selective marker gene; maintaining cells in selective media; and isolating viable cells containing a modified target sequence. 44. A method of supplying wild-type EHOC-l gene function to a cell which has a mutation/aneuploidy in the EHOC-l gene comprising introducing a wild-type EHOC-l gene or functional fragment thereof into said cell such that it is expressed. 45. Single strand DNA primers for amplification diagnosis of progressive myoclonus epilepsy, wherein said primers comprise a nucleic acid sequence derived from the nucleic acid sequence set forth as SEQ ID NO: 1, SEQ ID NO:5 or SEQ ID NO:6. 46. A method for detecting one or more EHOC-l alleles in a sample of nucleic acid comprising determining the presence or absence of variant nucleotide sequence in a gene contained in any of BAC clones.

Inventors: KORENBERG JULIE R, YAMAKAWA KAZUHIRO, KORENBERG, JULIE, R., YAMAKAWA, KAZUHIRO
Applicants: CEDARS SINAI MEDICAL CENTER, KORENBERG JULIE R, YAMAKAWA KAZUHIRO, CEDARS-SINAI MEDICAL CENTER, KORENBERG, JULIE, R., YAMAKAWA, KAZUHIRO
Application year: 1995
Main classification: C07K14
IPC codes: A01K67, A61K48, C07K14, C07K16, C12N15, C12Q1, C12R1, G01N33