CA2315268A1 - Oligophrenin-1, its expression product, and the diagnostic and therapeutic applications thereof - Google Patents

Abstract

The present invention relates to the identification of a new gene, called oligophrenin 1, its expression product, and the diagnostic and therapeutic applications of these nucleotide and peptide sequences.

Description

" A new gene called oligophrenin 1, its expression product, and the diagnostic and therapeutic -applications thereof".
The present invention relates to the identification of a new gene, called oligophrenin 1, and its expression product, as well as to the diagnostic and therapeutic applications of these nucleotide and peptide sequences.
A major challenge for human genetics is the identification of new causes of mental retardation, which, although present in about 3 °~ of individuals, is unexplained in over half of all cases. X-linked mental retardation is acknowledged to be a major cause of severe seaming difficulties. Surveys have shown an excess of males over females with severe mental retardation and later studies suggested that the excess was the result of an X-linked condition. X-linked mental retardation (XLMR) is a vastly heterogeneous group of disorders which can be roughly categorized as syndromic (MRXS) or non specific (MRX). Families with syndromic disorders usually have a quite distinct phenotypic presentation whereas families with non specific disorders present no distinctive somatic features. Despite recent advances in identifying genes such as FMR1 (Verkerk et .al., 1991 ), FRAXE (Knight et al., 1993 ; Gecz et al., 1996), L1-CAM (Vits et al., 1994), FGD1 and XH2 (Gibbons et al., 1995), involved in MRXS conditions, so far no gene significantly involved in MRX has yet been identified or cloned. Compilation of the literature and McKusick's catalogue data revealed at least 95 X-linked disorders in which mental retardation appears as the main feature. Of these 95, 40 have been regionally mapped on the X chromosome by conducting linkage studies using DNA
markers in single large families or in a collection of families with the same XLMR syndrome. Several loci appear to be located in the proximal Xq region.
However, it is impossible to evaluate how many MR genes there are in reality, partly because of the broad localisation and the presence of several overlaps between intervals of assignment. Thus, fine mapping and identification of genes implicated in nonspecific X-linked MR essentially depend on thorough investigation of molecular abnormalities such as balanced translocation, inversion or contiguous gene deletion associated with MR.

Bienvenu et al. (1997) have recently reported a molecular cytogenetic investigation of an X;12 balanced translocation observed in a female affected with a mild mental retardation and have localised the breakpoint in Xq12.
S The authors of the present invention have now cloned the gene responsible for MRX, which they have called the oligophrenin 1 gene.
In order to define the genomic structure of this gene, the authors of the present invention constructed and investigated a cosmidlphage contig that covers the gene. Determination of axon-intron boundaries was performed l0 through sequence comparison between cDNA clones and genomic DNA, which led to the identification of 25 axons.
The authors of the present invention have thus isolated and characterized the oligophrenin 1 transcripts. Said transcripts contain an open-reading frame (ORF) which is encoded by axon 2 to axon 24. This ORF is 15 2406 bases long and encodes a protein of 802 amino acids, called the oligophrenin 1 protein.
A subject of the present invention is thus an isolated nucleic acid having a sequence selected from the group consisting of sequence SEQ ID
n°
20 1 to SEQ ID n° 25, and a homologous nucleotide sequence thereof.
SEQ ID n° 1 represents the 5' fragment of the genomic DNA of the human oligophrenin 1 gene.
SEQ ID n° 2 to SEQ 1D n°25 represent fragments of the genomic DNA of the human oiigophrenin 1 gene including axons as shown in table 1.

Table 1 : identification of the sequences Sequence Identification exon inGuded in the fragment of the genomic DNA of the oligophrenin 1 gene SEQ ID n 2 exon 1 and exon 2 SEQ ID n 3 exon 3 SEQ lD n4 exon 4 SEQ ID n5 exon 5 SEQ ID n8 exon 8 SEQ ID n7 axon 7 SEQ ID n8 exon 8 SEQ ID n9 axon 9 SEQ ID n10 axon 10 SEQ ID n11 exon 11 SEQ ID n12 exon 12 SEQ ID n13 exon 13 SEQ ID n14 exon 14 SEQ ID n15 axon 15 SEQ ID n16 exon 18 SEQ ID n17 axon 17 SEQ ID n18 exon 18 SEQ ID n19 exon 19 SEQ ID n20 exon 20 SEQ ID n21 axon 21 SEQ ID n22 exon 22 SEQ ID n23 exon 23 SEQ ID n24 axon 24 SEQ ID n25 exon 25 SEQ ID n° 26 represents the cDNA fragment corresponding to the common open-reading frame (ORF).
A subject of the present invention is also an isolated nucleic acid having a sequence selected from the group consisting of axon 1 to exon 25 as identified in the sequence listing and in table 2, and a homologous nucleotide sequence thereof.

WO 99/31230 1'CTIEP98/08557 Table 2 : identification of axon sequences axon from nucleotideto nucleotide Sequence which n n includes said axon axon 1 1 634 SEQ ID n 2 axon 2 778 935 axon 3 403 498 SEa ID n 3 axon 4 483 544 SEQ ID n4 axon 5 451 522 SEQ ID n5 axon 8 416 517 SEA iD n6 axon 7 484 574 SEQ IO n7 axon 8 244 348 SEQ ID n8 axon 9 134 263 SEQ ID n9 axon 10 383 483 SEQ ID n10 axon 11 107 198 SEQ ID n11 axon 12 211 289 SEQ ID n12 axon 13 212 245 SEQ ID n13 axon 14 172 234 SEQ ID n14 axon 15 207 281 SEQ ID n15 axon 16 270 354 SEQ ID n18 axon 17 355 413 SEQ ID n17 axon 18 80 185 SEQ ID n18 axon 19 79 238 SEQ ID n19 axon 20 230 377 SEQ ID n20 axon 21 185 508 SEQ ID n21 axon 22 320 485 SEQ ID n22 axon 23 211 261 SEQ ID n23 ~

axon 24 115 156 SEQ ID n24 "A homologous nucleotide sequence" is understood as meaning a sequence which differs from the sequences to which it refers by mutation, insertion, deletion or substitution of one or more bases.
Preferably, such homologous sequences show at least 70 °~ of homology, preferably 80 °~ of homology, more preferably 90 °~ of homology to with any of sequences SEQ ID n° 1 to SEQ ID n° 26.

WO 99/31230 PCTIEP98/0855~
A polynucleotide of the invention, having a homologous sequence, hybridizes to the sequences to which it refers (any of sequences SEQ ID n° 1 to SEQ ID n° 26), preferably under stringent conditions.
Parameters that define the conditions of stringency depend upon the 5 temperature at which 50 °~ of annealed strands separate (Tm).
For sequences comprising more than 30 nucleotides, Tm is calculated as follows Tm = 81.5 + 0.41 (% G + C) + 16.6 Log (positive ion concentration) - 0.63 (%
formamide) - (6001polynucleotide size in base pairs) (Sambrook et al, 1989).
For sequences comprising less than 30 nucleotides, Tm is calculated as follows T",=4(G+C)+2(A+T).
Under appropriate stringent conditions avoiding the hybridization of non specific sequences, hybridization temperature is around from 5°C
to 30°C, preferably from 5°C to 10° C below the calculated Tm, and hybridization buffer solutions that are used are preferably solutions with high ionic strength, such as an aqueous 6 X SSC solution for example.
A nucleotide sequence homologous to the open-reading from SEQ ID n° 26 means a nucleotide sequence which differs from the sequences to which it refers by mutation, insertion, deletion or substitution of one or more bases, or by the degeneracy of the genetic code so long as it codes for a polypeptide having the biological activity of oligophrenin 1 protein, as defined below.
Said homologous sequences include mammalian genes coding for the oligophrenin 1 protein, preferably of primate, cattle, sheep, swine, or rodent, as well as allelic variants.
The nucleic acid sequences of the invention are useful for the detection of an abnormality, such as a mutation, in the oligophrenin 1 gene or in the transcripts of the oiigophrenin 1 gene. Such an analysis allows in vitro diagnosis of a neurological disorder associated with said abnormality.

A subject of the present invention is a method of in vitro diagnosis of a neurological disorder associated with an abnormality in the oligophrenin gene or in the transcripts of the oligophrenin 1 gene, wherein one or more mutation(s), preferably inducing a modification of the expression of the oligophrenin 1 gene is detected in the oligophrenin 1 gene or in the transcripts of the oligophrenin 1 gene.
A subject of the present invention is also a nucleic acid comprising a sequence identical to SEQ ID n° 26 or to homologous sequences thereof, except for a one base deletion of the nucleotide 1578 as shown in SEQ ID n° 26.
The present invention relates to methods of in vitro diagnosis wherein the nucleic acid sequences of the invention or probes or primers derived thereof are used to detect aberrant synthesis or genetic abnormalities at the oligophrenin 1 gene level.
The present invention is more particularly directed to a method of in vitro diagnosis comprising the steps of - contacting a biological sample containing DNA with specific oligonucleotides permitting the amplification of all or part of the oligophrenin 1 gene, the DNA contained in the sample having being rendered accessible, where appropriate, to hybridization, and under conditions permitting a hybridization of the primers with the DNA contained in the biological sample ;
amplifying said DNA ;
- detecting the amplification products ;
- comparing the amplified products as obtained to the amplified products obtained with a normal control biological sample, and thereby detecting a possible abnormality in the oligophrenin 1 gene.

The method of the invention can also be applied to the detection of an abnormality in the transcript of the oligophrenin 1 gene, by amplifying the mRNAs contained in a biological sample, for example by RT-PCR.
Thus another subject of the present invention is a method of in vitro diagnosis, as previously defined comprising the steps of - producing cDNA from mRNA contained in a biological sample ;
- contacting said cDNA with specific oligonucleotides permitting the amplification of al! or part of the transcript of the oiigophrenin 1 gene, under conditions permitting a hybridization of the primers with said cDNA ;
l0 - amplifying said cDNA ;
- detecting the amplification products ;
- comparing the amplified products as obtained to the amplified products obtained with a normal control biological sample, and thereby detecting a possible abnormality in the transcript of the oligophrenin 1 gene.
This comparison of the amplified products obtained from the biological sample with the amplified products obtained with a normal biological sample can be carried out for example by specific probe hybridization, by sequencing or by restriction site analysis.
A subject of the present invention is also a nucleic acid sequence which specifically hybridizes with a nucleic acid sequence of the invention as previously defined or with their complementary sequences.
"A sequence which specifically hybridizes [...J" is understood as meaning a sequence which hybridizes with the sequences to which it refers under the conditions of high stringency (Sambrook et al, 1989). Such sequences are preferably oligonucleotides having at least 15, and more preferably at least 20 bases.
3o Such sequences, which are useful as primers or probes for the diagnosis methods according to the present invention may be preferably selected from the group consisting of nucleic acid fragments of SEQ ID
N° 2 to SEQ ID N° 2fi as shown in table 3, or the complementary sequences thereof.

wo 99r~mo PcT~r9sosss7 Table 3 : identification of the oligonucleotide sequences Sequence which includes said Oli onucleotides oli onucleotide from nucleotide n to nucleotide n 727 746 SEQ m n 2 375 394 SEQ ID n 3 418 437 SEQ m n4 5s1 570 :123 445 SEQ iD n5 388 407 SEQ ID n6 5:40 559 436 458 SEQ m n7 219 239 SEQ ID n8 108 128 SEQ m n9 361 380 SEQ ID n10 81 100 SEQ ID n 11 188 207 SEQ ~ n 12 166 189 SEQ m n 13 '259 278 133 152 SEQ ID n14 151 170 SEQ m n15 221 244 SEQ ID n 16 305 324 SEQ ID n 17 25 44 SEQ m n 18 51 70 SEQ ID n 19 2~ 225 SEQ m n20 151 170 SEQ ~ n21 294 313 SEQ m n22 179 198 SEQ m n23 65 84 SEQ m n24 5~ ~6 SEQ m n 26 One skilled in the art very well knows the standard methods for analysing the DNA contained in a biological sample and for diagnosing a genetic disorder. Many strategies for genotypic analysis are available (Antonarakis et al., 1989, Cooper et al., 1991 ).
5 Preferably, one can use the DGGE method (Denaturing Gradient Gel Electrophoresis), or the SSCP method (Single Strand Conformation Polymorphism) for detecting an abnormality in the oligophrenin 1 gene. Such methods are preferably followed by direct sequencing. The RT-PCR method may be advantageously used for detecting abnormalities in the oligophrenin 1 10 transcript, as it allows to visualize the consequences of a splicing mutation such as axon skipping or aberrant splicing due to the activation of a cryptic site. This method is preferably followed by direct sequencing as well. The more recently developed technique using DNA chip can also be advantageously implemented for detecting an abnormality in the oligophrenin 1 gene (Bellis et I5 al., 1997).
The cloning of the oligophrenin 1 gene, as well as the identification of various mutations responsible for neurological disorders according to the invention, allow direct or semi-direct diagnosis. The specificity and reliability of such diagnosis methods are more particularly appreciable for prenatal diagnosis. The nucleic acid sequences of the present invention represent a highly interesting tool for genetic counseling.
The authors of the present invention have shown that the oligophrenin 1 protein is a rho-GAP protein and that the constitutional loss of oligophrenin 1 activity in humans results in cognitive impairment. Defects in the oligophrenin 1 gene, or in the oligophrenin 1 gene product may cause inactivation of the oligophrenin 1 protein, which leads to constitutive activation of its target GTPases.
Such constitutive activation of fio family members has been shown to produce marked changes in certain actin-based processes, to alter the cytoskeleton in cultured cells (Nobes et al., 1995) and to affect cell migration and axon outgrowth in vivo (Luo et al., 1994). In addition, constitutively active Rac1 rho-GTPase produced neuropathological changes in transgenic mice such as defects in axon outgrowth and dendritic spine morphogenesis in mouse Purkinje cells (Luo et al., 1996).
In the same manner, constitutive activation of a rho-GTPase resulting from the loss-of-function of oligophrenin 1 protein leads to a dysfunctioning of signal transduction pathways involved in cell migration and axon outgrowth during development of the nervous system. Mental retardation may be the clinical expression of such neuropathoiogical changes.
The oligophrenin 1 gene would thus be involved in disorders due l0 to an abnormal neurone migration. Such disorders include not only genetic disorders such as nonspecific X-linked mental retardation but also incurable cryptogenic epilepsies and neurodegenerative diseases, such as Alzheimer's disease and cognitive impairments related to aging.
The present invention also provides transgenic non-human animals or cells thereof.
Said transgenic animal can have a exogenous oligophrenin 1 protein of this invention due to the presence of a gene encoding and expressing that protein or part of that protein.
Transgenic animals are generally well known, as are their methods of production.
The present invention contemplates a non-human animal containing a oligophrenin 1 gene of the present invention integrated in the genome of the animal's somatic and germ cells, i.e., a transgenic animal;
preferably transgenic mammals.
Animals containing a transgene encoding a oligophrenin 1 protein of the present invention are typically prepared using the standard transgenic technology described in Hogan et al. (1987) and Palmiter et al.
(1986). Production of tra~sgenic mammals is also possible using the homologous recombination transgenic systems described by Capecchi (1989).
Preparation of transgenic mammals has also been described in WQ94/21670.

One technique for transgenically altering a mammal is to microinject a rDNA into the male pronucleus of the fertilized mammalian egg to cause one or more copies of the rDNA to be retained in the cells of the developing mammal.
Alternative methods for producing a non-human mammal containing a rDNA of the present invention include infection of fertilized eggs, embryo-derived stem cells, totipotent embryonal carcinoma (Ec) cells, or early cleavage embryos with viral expression vectors containing the rDNA (Palmiter et al., (1986)).
A transgenic animal can also have a mutation in its own native oligophrenin 1 gene, thereby rendering the oligophrenin 1 protein non-functional (i.e., a "knockout" trangenic animal). Such an animal is useful as it presents the clinical conditions associated with the defects in the mutated oligophrenin 1 protein, and further can be a model for evaluation of candidate therapeutics that would treat subjects with defects in that protein.
More particularly, transgenic non-human animals or cells in culture, that overexpress oligophrenin 1 protein or preferably express a native oligophrenin 1 protein that has been rendered non-functional ("knock-out"
transgenic animal) may be useful in a method for screening chemical entities or drugs likely to act on the signaling pathway to which the rho-GAP""R"
protein (oligophrenin) belongs.
Transgenic non-human animals or cells thereof that overexpress oligophrenin 1 protein refer to animals or cells thereof that express an exogenous oligophrenin 1 protein of the invention in addition not the native protein.
In one embodiment, the screening method of the invention comprises the steps of i) administering a drug to be tested to a transgenic non-human animal that overexpress oligophrenin 1 protein or preferably express a native oligophrenin 1 protein that has been rendered non-functional ; and ii) observing clinical expression of neuronal changes in vivo andlor in vitro culturing nervous cells from said animal and observing the stimulation or recovery of axon outgrowth or morphogenesis.
In another embodiment, the screening method of the invention comprises the steps of i) contacting a drug to be tested with nervous cells or nervous tissue cultures that overexpress oligophrenin 1 protein or preferably express a native oiigophrenin 1 protein that has been rendered non-functional ; and ii) measuring the axon outgrowth.
In the above embodiment, said cells are either obtained from said transgenic animals or are established cell lines, such as neuroblastoma or primary cultures of neuronal cells, which have been transfected by a DNA
construct, e.g by means of a viral vector, allowing the expression of exogenous oligophrenin 1 protein or rendering the native oligophrenin 1 non functional.
Drugs selected by the methods of screening as above-defined, and pharmaceutical compositions containing such a drug in association with a pharmaceutically acceptable carrier, are also encompassed by the present invention.
2o The ORF of the oligophrenin 1 gene as shown in SEQ ID n° 2fi according to the invention encodes a protein of 802 amino acids with a relative molecular mass of 91 kD. Hydropathy analysis (Kyte and Doolittle, 1982) suggests that the oligophrenin 1 protein is hydrophilic. Based on consensus motifs in PROSITE database (Bairoch et al., 1997), many potential phosphorylation sites were predicted includingvra tyrosine kinase phosphorylation site at position 142. Comparison of the protein sequence with other sequences in the databases indicated that the oligophrenin 1 gene encodes a fio-GAP containing protein.
Sequence alignment shown in figure 3b illustrates the remarkable similarity between the predicted oligophrenin 1 domain and other members of the rho-GAP subfamily. This similarity extends over 180 residue region localised in the central part of the predicted protein and concerns the three structurally conserved regions (SCRs) that are specific to the rho-GAP
proteins.
Among rho-GAPs, the oligophrenin 1 protein showed the greatest similarity to the chicken Graf protein (Hildebrand et al., 1996). This similarity extends on both sides of the rho-GAP domains, but oligophrenin 1 does not contain the SH3 domain reported for the Graf protein. The rho-GAP activity of the oligophrenin 1 protein is consistent with the functional analysis of the chicken Graf protein, which has both part of the N-terminal and rho-GAP
domains identified in the oligophrenin 1 protein. Graf protein has been shown to preferentially stimulate the GTPase activity of the GTP-binding proteins RhoA and Cdc42 (Hildebrand et al., 1996).
The C-terminal part of oligophrenin 1 protein does not match any known sequence, whereas the N-terminal domain of oligophrenin 1 protein is similar to a highly conserved protein, of unknown function, identified in C.
elegans, mouse and human (Fig. 3c). This protein presents two isoforms identified as CELZK328 and CELT04C95 (Genbank), which correspond to two different ORF.
A further subject of the present invention is thus an isolated oligophrenin 1 polypeptide substantially comprising the aminoacid sequence of SEQ ID n° 27 or a homologous aminoacid sequence thereof.
The above term "substantially" is understood as meaning that said isolated oligophrenin 1 polypeptide exhibits the same biological andlor immunological properties, as the native oligophrenin 1 protein.
More particularly said aminoacid sequence may be SEQ ID n° 26, or a homologous aminoacid sequence thereof.
"A homologous aminoacid sequence" is understood as meaning a sequence which differs from the sequences to which it refers by mutation, insertion, deletion or substitution of one or more aminoacids, without inducing modification of biological andlor immunoiogical properties. Said derivative WO 99/311.30 PCT/EP98/08557 aminoacid sequence shows at least 60 °~ of homology, preferably 70 % of homology, preferably 80 % of homology with the oligophrenin 1 polypeptide of SEQ ID n° 26.
The "biological properties" of the poiypeptides of the invention 5 refer to the activity of the oligophrenin 1 protein, which enhances GTPase activity of small Ras-like GTPases and hence turns them off.
The "immunological properties" of the polypeptides of the invention refer to the ability of the polypeptides of the invention to induce an to immunological response mediated by antibodies which recognize the oligophrenin 1 polypeptide of the invention.
The polypeptides according to the invention can be obtained by any of the standard methods of purification of soluble proteins,' by peptide 15 synthesis or by genetic engineering. Said techniques comprise the insertion of a nucleic acid sequence coding for a peptide of the invention into an expression vector, such as a plasmid, and the transformation of host cells with the expression vector, by any of the methods available to the skilled person, like for instance electroporation.
The present invention thus relates to vectors for cloning andlor expression comprising a nucleic acid sequence of the invention and to host cell transfected with these vectors. The expression vector according to the invention comprises a nucleic acid sequence encoding a polypeptide of the invention, said nucleic acid sequence being operably linked to elements allowing its expression. Said vector advantageously contains a promoter sequence, signals for initiation and termination of translation, as well as appropriate regions for regulation of translation. Its insertion into the host cell may be transient or stable. Said vector may also contain specific signals for secretion of the translated protein.
These various control signals are selected according to the host cell which may be inserted into vectors which self-replicate in the selected host cell, or into vectors which integrate the genome of said host.

Host cells may be prokaryotic or eukaryotic, including but not limiting to bacteria, yeasts, insect cells, mammalian cells, including cell lines which are commercially available.
A subject of the present invention is also a method for producing a recombining oligophrenin 1 polypeptide, wherein said host cell is transfected with said expression vector and is cultured in conditions allowing the expression of a polypeptide according to the invention.
The present invention also relates to monoclonal or polyclonal antibodies, or fragments thereof, or chimeric or immunoconjugate antibobies, which are capable of specifically recognizing a polypeptide according to the invention.
Polyclonal antibodies can be obtained from serum of an animal immunized against the oligophrenin 1, which can be produced by genetic engineering for example, as above described, according to standard methods well-known by one skilled in the art.
Monoclonal antibodies can be obtained according to the standard method of hybridoma culture (Kohler and Milstein, 1975).
The antibodies of the present invention can be chimeric antibodies, humanized antibodies, or antigen binding fragments Fab and F(ab')2. They can also be immunoconjugated or labelled antibodies.
Said antibodies are particularly useful for detecting or purifying a oligophrenin 1 polypeptide according to the invention in a biological sample.
They are more particularly useful for detecting an abnormal expression of the oligophrenin 1 protein in connection with neurological disorders as above described.
Another subject of the present invention is a pharmaceutical composition comprising a purified oligophrenin 1 polypeptide of the invention andlor a homologous poiypeptide thereof, in association with a pharmaceutically acceptable carrier.

A further subject of the present invention is a pharmaceutical composition comprising a nucleic acid encoding said polypeptides and a pharmaceutically acceptable carrier. Said nucleic acid, preferably inserted in a vector, may be administered in a naked form or in association with transfection facilitating agents.
A further subject of the invention is a pharmaceutical composition comprising an anti-sense sequence capable of specifically hybridizing with a nucleic acid sequence encoding said polypeptides, in association with a pharmaceutically acceptable carrier.
A still further subject of the invention is a pharmaceutical composition comprising an antibody directed against said polypeptides, in association with a pharmaceutically acceptable carrier.
Preferably the present invention is directed to a pharmaceutical composition comprising a purified oligophrenin 1 polypeptide of the invention andlor a homologous polypeptide thereof, in association with a pharmaceutically acceptable carrier.
The term "homologous pofypeptide", as active ingredient of a pharmaceutical composition, refers to a polypeptide with a homology of at least 40 °~, preferably of at least 60 % in comparison to the oiigophrenin 1 protein.
Such homologous polypeptides include any known protein which exhibits a rho-GAP activity. Preferably said homologous polypeptide is for example the protein CELZK328 or CELT04C95.
The pharmaceutical compositions of the invention are useful for preventing and/or treating neurological disorders, wherein the oligophrenin 1 protein. or a homologous protein thereof is implicated. As above underlined, the authors of the present invention have shown that defects in a Ras-like GTPase (rho-GAP) dependent signalling pathway are associated with cognitive impairment, resulting from misguided axon growth and/or defective cell migration. Consequently, the disorders which are more particularly aimed at are disorders of the central nervous system in connection with the axons!

development, more particularly a disorder associated with cognitive impairment. Such disorders include nonspecific X-linked mental retardation, as well as cryptogenic epilepsies or neurodegenerative diseases, such as Alzheimer's disease and cognitive impairments related to aging.
Another subject of the invention is the use of a purified oligophrenin 1 polypeptide of the invention andlor a homologous polypeptide thereof, , in association with a pharmaceutically acceptable carrier for the manufacture of a medecine for preventing and/or treating neurological l0 disorders, wherein the oligophrenin 1 protein or a homologous protein thereof is implicated.
The pharmaceutical compositions of the invention may be administered to a mammal, preferably to a human, in need of a such treatment, according to a dosage which may vary widely as a function of the age, weight and state of health of the patient, the nature and severity of the complaint and the route of administration.
The appropriate unit forms of administration comprise oral forms such as tablets, gelatin capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, subcutaneous, intramuscuiar, intravenous, intranasal or intraoccular administration forms and rectal administration forms.
A further subject of the present invention is a method of preventing andlor treating neurological disorders resulting from defects in the oligophrenin 1 gene or in the oligophrenin 1 protein or in a homologous gene or protein thereof, which comprises administering to a subject in need of a such treatment an amount of a pharmaceutical composition as above defined effective to prevent andlor alleviate said neurological disorders.
The present invention is further illustrated by, but not limited to, the figures and the examples that follow.

LEGENDS TO FIGURES
Figure 1. (a) Physical map of the Xq12 focus and genomic structure of the oligophrenin 1 gene spanning the X-chromosomal breakpoint. YAC, PAC and cosmid contigs are indicated with lines and STSs by vertical bars. The STS
C16T3 was generated from the distal end of the 9 kb Hindlll fragment containing the X-chromosomal breakpoint (represented by an arrow). The oligophrenin 1 gene spans at least 300 kb and it consists of 25 axons of which 23 are coding (white boxes). (b) Southern blot analysis of Hindlll digested genomic DNAs from the patient exhibiting the (X;12) translocation and a normal female using the STS C16 T3 as probe. The junction fragment is indicated by JF.
Figure 2. Fetal and multiple adult tissue northern blots containing poly(A)+
RNA (Clontech~) hybridized with C2 cDNA probe. A 7.5 kb transcript was observed after an overnight exposure at - 80°C. Hybridization of Northern blots with an actin probe was performed to assess differences between 2o amounts of loaded poly(A)+ RNA samples.
Figure 3. (a) Coding part of the cDNA and deduced amino acid sequences of the oligophrenin 1 gene. Nucleotides in bold con-espond to axon-axon boundaries. The GAP domain is underlined, Primers used in RT-PCR to study the expression of the gene in the patient with the X;12 translocation are double underlined and the internal primer used for hybridization to ascertain PCR products is shown in dotted line. The deleted nucleotide at position 1578 is indicated in italic bold case. (b) Sequence alignment of the GAP domain present in oligophrenin 1 and different rhoGAP proteins reviewed in Aelst et 3o al., 1997. The GAP domain contains three Structurally Conserved Regions (SCRs) (Boguski et al., 1993). CELZK328 corresponds to an ORF predicted from sequences of the C.elegans genome. (c). Sequence alignment showing the high conservation of oligophrenin 1 N-terminal domain. CELT04C95 corresponds to another C. elegans ORF, different from CELZK328.
EST483210 is part of the mouse homologue oligophrenin 1 cDNA and EST387042 corresponds to a human EST that was localized on chromosome 5 11 by the Whitehead Institute.
Identical residues are indicated by black shading, similar residues by grey.
The alignment was performed using Multalin (Corpet et al., 1988) and Boxshade softwares.
10 Figure 4. (a). Study of oligophrenin 1 transcript in the patient with the (X;12) translocation and in a normal female used as control (XX). Southern blot of PCR products amplified from total RNAs isolated from EBV transformed lymphoblastoid cell lines.
(b) Nucleotide sequences showing the one base pair deletion. Direct 15 sequencing of PCR products corresponding to axon 19 of IV-3 proband.
(c). Segregation analysis of the mutated allele in XLMR family D. Open squares, unaffected males; closed squares, affected males; open circles, unaffected females; dotted symbols, phenotypically normal carrier females.

EXAMPLE 1 : Identification of the oligophrenin 1 gene 1. Experimental procedures s Case report and family materials Clinical data and diagnosis concerning the female patient with the t(X;12) translocation were previously described (Luo et al., 1996).
Concerning the MRX families, a linkage study was reported in The European XLMR Consortium's report (1997).
YAC and PAC clones YAC clones of the Xq12 locus were obtained from the UK HGMP
Resource Centre. PAC clones were obtained from the German resource center (RZPD). Primer sequences corresponding to STSs are available in Genome Data Base. STSC16T3 is a 189 by fragment amplified with the primers:
C16T3F (5' CACAGCAAGCAATAAGCACT 3') and C16T3R (5' TGTCTCCTGTGCTCTTTCCA 3'}. Overlaps between clones and STS mapping were performed by a combination of STSIEST amplification and hybridization approaches.
cDNA isolation Approximately 1 X 106 recombinant clones of a Igt10 human fetal brain cDNA library (Clontech) were plated and screened following standard techniques (Sambrook et al., 1989}. Library screening was performed using as probe RT-PCR products generated with primers located within the predicted axons. Positive clones were plaque purified and their inserts were subcloned into bluescript vector and sequenced. 3' RACE PCR (Clontech kit) was used to obtain the 3' end of the cDNA. The full-length cDNA is a composite of 14 clones (only 4 clones are represented on figure 1a).

Genomic DNA sepuencina of human cosmid clones Human cosmid clones were detected in the Imperial Cancer Research Fund (ICRF) flow-sorted human X chromosome library (Nizetic et al., 1991 ) using B2 cDNA as a probe. Cosmid 2C6, 4D2 and 35 shown in figure 1 a are from a cosmid library corresponding to the YAC 4690. Exon-intron boundaries were identified through sequence comparisons between cDNA and genomic DNA clones. To generate genomic sequences, DNAs of cosmid clones were used as templates and primed with exonic oligonucleotides. ICRF
coordinates of cosmid clones shown in figure 1 are as follow: cos12:
ICRFc104J1515Q8, cos15: ICRFc104K1628Q8, cos7: ICRFc104P0212Q8, cos3: ICRFc104B1719Q8, cos11: ICRFc104F178Q8, cosy:
ICRFc104B1515Q8.
2. Results As a first step in identifying a potential gene in Xql2 involved in MRX, Bienvenu ef al. (1997) have reported a molecular cytogenetic investigation of an X;12 balanced translocation observed in a female affected with a mild mental retardation. The Xq breakpoint was localized within an ICRF
850 kb YAC clone y900H0493 (4690), positive for PGKP1 and DXS159 markers.
The authors of the present invention have used long-range restriction maps of the YAC clone 4690, FISH analyses and somatic hybrid cell lines containing the derivative chromosome 12 as their only human X-chromosomal component to regionally fine map the X chromosomal breakpoint. Figure 1 a depicts the location of the translocation breakpoint on the normal X and summarizes YAC, PAC and cosmid contigs spanning the breakpoint. The probe, STSC16T3, which detects the junction fragment and localized the breakpoint to a 9 kb Hindlll fragment (fig 1 b) was isolated from the cosmid clone 4D2 (Fig 1 a). Aberrant bands confirming the latter results were also obtained by hybridization of the same probe to a Southern blot containing DNA from the patient digested with several other enzymes.
Sequencing of randomly subcloned Hindlll fragments including the 9 kb fragment isolated from cosmid clones spanning the breakpoint, and searches for homology in data bases revealed sequence identities between the isolated sequences and those corresponding to the PAC clone 360E18 (Fig 1 a) which were generated by the Sanger Centre (Cambridge, UK).
Available sequences were then used for computational analyses and comparison with nucleotide and protein sequences. Some of the potential axons identified by GRAIL (Kel et al., 1993) and FEXHIHEXON (Lerman et al., 1987) programs showed a significant homology with the mouse EST 483210.
The predicted polypeptide corresponding to this EST revealed a significant homology with the human EST 387042 localised on chromosome 11 and with the C. elegans ORF CELT04C95. Further investigations suggested that the C.
elegans ORF is represented on the genomic sequences derived from the PAC
clone by 8 different potential axons scattered over 130 kb (Fig. 1 a). These predictions were confirmed by RT-PCR experiments using primers located within the potential axons and human fetal brain total RNA. Furthermore, hybridization of the RT-PCR products to a Northern blot containing polyA+
RNA detected a 7.5 kb transcript most highly expressed in fetal brain (Fig.
2).
Together, these results indicated the presence of a candidate gene located in the vicinity of the transiocation breakpoint.
In order to identify the full length cDNA the authors of the present invention used a combination of fetal brain cDNA library screening, PCR and rapid amplifications of cDNA ends (RACE). This approach enabled to obtain a composite full length nucleotide sequence of the cDNA (Figure 3a).
To confirm that the identified gene is disrupted by the translocation breakpoint the structure of the gene including the axon-intron boundaries was determined through sequence comparisons between cDNA
and cosmid genomic DNA clones isolated either from a cosmid library generated from YAC clone 4690 or the ICRF flow sorted X-specific cosmid WO 99/31230 PCT/EP9810855?

library (fig 1 a). The physical mapping of the 25 axons allowed to demonstrate that the candidate MRX gene is transcribed from telomere to centromere and the translocation breakpoint maps within the second intron leading therefore to a displacement on the derivative chromosome 12 of the first two axons including the one containing the putative translation initiation codon.
Confirmation of this latter result was obtained by FISH analysis using as probe a cosmid clone containing the first two axons of the gene (cps 12, fig 1a).
This experiment showed that this cosmid maps exclusively on the derivative chromosome 12 (data not shown).
l0 To investigate the gene expression and examine whether both alleles are inactive, RT-PCR experiments were performed on RNA isolated from EBV-transformed lymphoblastoid cell lines (t_CL) of the patient and a female control using primers located in axons 1 and 2. These experiments failed to amplify a normal gene product on RNA of the patient (fig 4a).
Consistently, the normal X chromosome was found to be late replicating, indicating preferential X-inactivation of the normal X chromosome (Bienvenu et al., 1997), and the MRX candidate gene was found to undergo X-inactivation.
EXAMPLE 2 : Identification of mutations in MRX families 1. Experimental procedure Mutations analysis Genomic DNA was extracted from EBV - transformed lymphoblastoid cell lines using standard protocols. The 23 coding axons and axon-intron boundaries were individually amplified with specific primers. In each amplification one primer was a 5' psoralen-modified primer (Fernandez et al., 1993). PCR products were checked on standard agarose gels prior to analysis by the DGGE technique. When an aberrant pattern of migration was observed, the corresponding PCR product was purified and directly sequenced on both strands using the Dye Terminator Cycle Sequencing kit protocol (Applied Biosystems).
Figure 4(a) represents a Southern blot of PCR products amplified from ,total RNAs isolated from EBV transformed lymphoblastoid cell lines. RT
5 PCR were performed with (+) or without (-) reverse transcriptase and cDNAs were amplified for 40 cycles with primers located in axon 1 and 2 (figure 3, double underlined nucleotide sequences). After gel electrophoresis, the Southern blot was hybridized with an internal oligonucieotide (dotted line on figure 3). The 650 by fragment corresponds to an amplification from the 10 contaminating genomic DNA (intron 1 is 140 by long). The lane labelled DNA, corresponds to the PCR product obtained from a female total genomic DNA.
The negative control indicated by Ct(-) corresponds to a PCR experiment without template. RT-PCR amplification of the ubiquitously expressed transcript produced by the distal part of the dystrophin gene was used as 15 internal standard.
The mutation to co-segregate with the mental retardation phenotype as shown on figure 4c was detected by denaturing gradient gel. electrophoresis of PCR products corresponding to axon 19 of the oligophrenin 1 gene. Exon 19 was amplified by PCR with primers 19F (5' GTT AAT CTT GCC CCT TTT CT
20 3') and 19R (5' Psoralen- TA GGA AGA CAG GTA GTG AGA AT) yielding a 221 by product. 10 NI of each amplified product was mixed with 10 pl of normal control PCR product. Heteroduplexes were generated by denaturing for 10 min, and subsequent reannealing for 45 min at 56°C. The samples were electrophoresed through a 25-65°r6 denaturant 6% polyacrylamide gel for 7.5h 25 at 60°C and 160V. The characteristic shifted profile displayed by the mutated allele allow an easy study of the familial segregation.
2. R-Its In order to prove that the isolated gene is responsible for non-specific mental retardation, four unrelated probands from MRX families (The WO 99!31230 PCT/EP98/08557 European XLMR Consortium, 1997), previously mapped in genetic intervals which encompass the candidate gene, were analysed for the presence of point mutations. The strategy involved investigation by DGGE (Lerman et al., 1993) (denaturing gradient gel electrophoresis) of PCR products corresponding to all coding axons and sequencing of axons exhibiting abnormal migration profiles.
PCR primers were designed not only to amplify individual axons but also sequences flanking the axons. DGGE analyses of amplified products, corresponding to axon 19, from the proband IV-3 of the family D (fig 4c) showed an abnormal shift in mobility. Compared with the normal product, l0 sequence analysis showed that the aberrant product contained a one base pair deletion of the nucleotide 1578 (fig 4b); the resulting frameshift mutation was predicted to cause premature translation termination four codons downstream of the mutation. Cosegregation of the mutation with the disease which was confirmed in the large family using the DGGE technique (fig 2b) and the absence of this mutation in 100 control individuals indicate that the deletion does indeed cause the mutant phenotype.
EXAMPLE 3 : Expression of the oligophrenin 1 gene C2 cDNA clone was isolated from a fetal brain cDNA library. C2 DNA probe was used to hybridize poly(A)' RNA (Clontech~) contained in fetal and multiple adult tissues. A 7.5 kb transcript was observed after an overnight exposure at - 80°C. Hybridization of Northern blots with an actin probe was performed to assess differences between amounts of loaded poly(A)+ RNA
samples.
As shown in figure 2, the oligophrenin 1 transcript was most abundant in RNA from fetal brain. A lower level of expression was also detected in several other tissues including adult brain. To further investigate 3o the distribution of the transcript during development, in situ hybridization was used to examine the expression of the mouse homologous gene in embryonic days (E) 10.5, E12.5, E14, E18 and in postnatal day 1 of mouse embryos and postnatal tissues. In addition to a low expression in all tissues with no significant differences, it was found that the gene is expressed at a higher level in all parts of the developing neuroepithelium of the neural tube. During later differentiation stages and in the mature brain a significant level of expression is visible in different structures of the brain with no striking distribution of the mRNA expression.
Several lines of evidence show that defects in oligophrenin 1 are responsible for X-linked non-specific mental retardation. First, the oligophrenin 1 gene maps to a potential mental retardation genetic locus in Xq12 identified by linkage analyses (tubs et al., 1996, The European XLMR Consortium, 1997). Second, literature reports (Davies, 1997) concerning two patients with complete androgen insensitivity syndrome (CAIS) and mental retardation showed the presence of deletions which include several markers both proximal and distal to the AR gene and extend to DXS905 and DXS908. The above mapping data showed that these markers are located within the second and fifth intron of the oligophrenin 1 gene (Fig. 1a), demonstrating therefore that most of the axons are deleted in these two patients with mental retardation.
In contrast, deletions in two CAIS patients without mental retardation do not extend to the oligophrenin 1 gene as deletions are limited to the androgen receptor gene itself (Davies, 1997). Third, investigation of this gene in the female patient with mental retardation and a t(X;12) demonstrated an absence of expression of both alleles resulting from the disruption of one allele by the translocation breakpoint and a preferential inactivation of the second allele.
Fourth, a one base pair deletion within the rho-GAP domain, predicted to result in a severe abbreviated and nonfunctional oligophrenin 1 protein, cosegregates with a recessive mental retardation phenotype in a large affected family mapped to the pericentromeric region. Finally, the oligophrenin 1 mRNA
is highly expressed in fetal brain, a finding consistent with the disease phenotype.

REFERENCES
1. Aelst, L.V. & D'Souza-Schorey, C. Rho GTPases and signaling networks.
Genes & Dev. 11, 2295-2322 (1997).
2. Bairoch, A., Bucher, P. & Hofmann, K. The PROSITE database, its status in 1997. Nucleic Acids Res 25, 217-21 (1997).
3. Bienvenu, T., et al. Mapping of the X-breakpoint involved in a balanced X;12 translocation in a female with mild mental retardation. Eur. J. of Hum Genet 5, 105-109 ( 1997).
4. Boguski, M.S. 8~ McCormick, F. Proteins regulating Ras and its relatives.
Nature 366, 643-54 ( 1993).
5. Capecchi et al., Science 244, 1288-1292 (1989).
6. Corpet, F. Multiple sequence alignment with hierarchical clustering.
Nucleic Acids Res 16, 10881-90 ( 1988).
7. Davies, H. R., et al. Androgen insensitivity with mental retardation: a contiguous gene syndrome ? J Med Genet 34, 158-60 (1997).

8. Fernandez, E., et al. Use of chemical clamps in denaturing gradient gel electrophoresis: application in the detection of the most frequent Mediterranean beta-thalassemic mutations. PCR Methods Appl 3, 122-4 (1993).

9. Gecz, J., et al. Cloning and expression of the murine homologue of a putative human X-linked nuclear protein gene closely linked to PGK1 in Xq13.3. Hum Mol Genet 3, 39-44 (1994).

10. Gedeon; A. K., Donnelly, A.J., Mulley, J.C., Kerr, B. & Turner, G. How many X-linked genes for non-specific mental retardation (MRX) are there? [letter).
Am J Med Genet 64, 158-62 (1996).

11. Gecz J. et al., Identification of the gene FMR2, associated with FRAXE
mental retardation. Nature Genet 13, 105-108 (1996).

12. Gibbons RJ et al., Syndrome mental retardation due to mutations in a regulator of gene expression. Hum Mol Genet 4, 1705-1709 (1995).

13. Herbst, D.S. & Miller, J.R. Nonspecific X-linked mental retardation II:
the frequency in British Columbia. Am J Med Genet 7, 461-9 (1980).

14. Hildebrand, J.D., Taylor, J.M. & Parsons, J.T. An SH3 domain-containing GTPase-activating protein for Rho and Cdc42 associates with focal adhesion kinase. Mol Cell Biol 16, 3169-78 (1996).

15. Hogan et al., Cold Spring Harbor, NY Manipulating the mouse embryo ; a laboratory manual (1987).

16. Kel, A.E., et al. SITEVIDEO: a computer system for functional site analysis and recognition. Investigation of the human splice sites. Comput App! 6iosci 9, 617-27 (1993).

17. Knight SJI et al., Trinucleotide repeat amplification and hypermethylation of CpG islands in FRAXE mental retardation. Cell74, 127-134 (1993).

18. Kozak, M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44, 283-92 ( 1986).

19. Kyte, J. & Doolittle, R.F. A simple method for displaying the hydropathic character of a protein. J Mol Biol 757, 105-32 (1982).

20. Lamarche, N. & Hall, A. GAPs for rho-related GTPases. Trends Genet 10, 436-40 ( 1994).

21. Lerman, L.S. & Siiverstein, K. Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods Enzymol 155, 482-501 (1987).

22. Lubs, H.A., et a/. XLMR genes: update 1996. Am J.Med Genet 64, 147-57 (1996).

23. Luo, L., Liao, Y.J., Jan, L.Y. & Jan, Y.N. Distinct morphogenetic functions of similar small GTPases: Drosophila Drac1 is involved in axonal outgrowth and ~myoblast fusion. Genes Dev 8, 1787-802 (1994).

24. Luo, L., et al. Differential effects of the Rac GTPase on Purkinje cell axons and dendritic trunks and spines. Nature 379, 837-40 (1996).

25. Mermod, N., O'Neill, E.A., Kelly, T.J. & Tjian, R. The proline-rich transcriptional activator of CTFlNF-I is distinct from the replication and DNA
binding domain. Cell58, 741-53 (1989).

26. Morrissey, J., et aL A serinelproline-rich protein is fused to HRX in t(4;
11 ) acute leukemias. Blood 81, 1124-31 (1993).

27. Nobes, C.D. & Hall, A. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress 5 fibers, lamellipodia, and filopodia. CelI81, 53-62 (1995).

28. Nizetic, D., et al. Construction, arraying, and high-density screening of large insert libraries of human chromosomes X and 21: their potential use as reference libraries. Proc Natl Acad Sci U S A 88, 3233-7 (1991 ).

29. Palmiter et al., Ann Rev Genet 20, 465-499 (1986).
l0 30. Sambrook, J., Fritsch, E. F. & Maniatis, T. Molecular cloning: A
laboratory manual, Cold Spring Harbor Laboratory Press ( 1989).
31. Solovyev, V.V., Salamov, A.A. & Lawrence, C.B. Predicting internal axons by oligonucleotide composition and discriminant analysis of spliceable open reading frames. Nucleic Acids Res 22, 5156-63 (1994).
15 32. The European XLMR Consortium. X-linked non specific mental retardation (MRX): linkage studies in 25 unrelated families. 8th International Workshop on the Fragile X and X Linked Mental Retardation. Picton; Ontario, Canada, 17-22 August 7997. (1997).
33. Verkerk AJM et al., Identification of a gene (FMR-1 ) containing a CGG
20 repeat coincident with a breakpoint cluster region exhibiting length variation in fragile X syndrome. .Cell 74:905-914 (1991 ).
34. Vits L. et al., MASA syndrome is due to mutations in the neural cell adhesion gene LtCAM. Nature Genet7, 408-413 (1994).
35. Zipkin, LD., Kindt, R.M. & Kenyon, C.J. Role of a new rho family member in 25 cell migration and axon guidance in C.elegans. CeI190, 883-894 (1997).

SEQUENCE (L1]LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT:
(A) NAME: Institut National de la Sante et de la Recherche Medicale (B) STREET: 101 rue de Tolbiac (C) CITY: Paris (E) COUNTRY: FRANCE
(F) POSTAL CODE (ZIP): 75013 (ii) TITLE OF INVENTION: oligophrenin 1 gene and protein (iii) NUMBER OF SEQUENCES: 27 (iv) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0, Version #1.25 (EPO) (2) INFORMATION FOR SEQ ID NO: 1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1650 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 1:
TTTTTCAACC CATGGTGCAT TTAAAGCTCA TTTTGGACAT TTTCCCATAA ATTAACCTTA
AAGATAAAAA GAGTAAGAAA CAAAACTTTC CCCTGAGATG TGGCTACTTA TTTATTTTCA

GAGGGCGTTT TCTCATTGCC ATTCTGCTAT ATAGTGTAGT GGTCAAGAGC ACTACCTCTA

GAGCCAGCCA GGCTGGGCTC AAGTTCAAGT GCTGCCATTT AACTAGCTGT TTGTCCTTCG

GCAAGTCACT TAAACTCTCT TTGACCCAGC TTCTCCATCT TTAAATGGGT ATAATAATAA

AACCATCCTC ATAGGGTTGT TTTGAAGATT AGTGAGATGG GCGATAGGTT GTGTGGTGGG

TAGAATAATG TTTCCTCCTT CACAGATGTC CATGTCCTGT CCTGAAACCT GTGGCTACGT

TATCTTATGT GGCAAAATGA AATTTGCAGA TGTGATTAAG GATGATGTGA TGGGGGAGAT

TATCCTGTGG ACCCAGTGTA ATCATAAGGG TCCTTAAGGG GAGGCATGAG AATCACAGTG

ATGTAGCATG AGAAAGACTT GACTGGCCAG TGCTAGCTTT AAAGACGGAG GAAGGGAGCT

ATGAGCCAAA GGATGTGAGC AGCTTGAGCC GGAAAAGGTA AGGAACCAGA TTCTACCCCT

AGACTATCTG AAAATGAACA CAACCCTTCT AACATCTTGA TTTGATCCCA GTGAGGCCCA

TTTCTGATTT CTGCCTTCTG GAACTGTAGA TTATAACTTT ATGCTGATTT AGGACATCAA

GTTTGTGGTA ATTTGTCACA GCAGGAGCAA CAGGACCTAA TTCAGGTGGG TCAGCCACTA

TTATTCTATT TACAACCCCC TTCCGTTCTG GCTTTCTTCA CAGGGCAATC TAATGTAAAC

ACTTCTGCAG AATTAAAGGG ATCTCTTTGC CCACGCACAA ACTATTTTTT AGTTTTTCGA

GCCTCTACCT ACCCTTAGTC TCAAGACAGC TGCCTTTAAG GTAAGCTGCA AGACAATTTA

TAACATCCTA TTCTTAGACT TTCCACTTAC ACATGGAAAA TCATATGCAT TATCGGCGTC

TGCCACAAAA GCCATGCTCT GAGGTCCTAT TCAGGAAAAT ATTAACTCTG AATTATGGCC

TCCCTTTCTT TGGCAGGGGG CACTGAGCTT CACGGAGACC CTAGAGTAGG AATCAACCCT

TCAGGTGGAA GGCCCAAGGG AATTAGAGCA CTTAGAGCAG GAGACATGGG AAAAGAAAGA

GGAAAAGAAG TTGGTAGGTC CGAAGGAGGC TGGAAAGAGG GGAAGGCAAA AGAATACACA

ACCCAGCCGA ATCATGGGAA GTGAGAGATG GCTTCCTGCA ATCTAGTCTT GCGGGTTCTT

GCTCTAGTTT CGGAAACTTC CCCCGCAGAG TCCAGCGCCG CGCACCTGCC CCTTGGGGCC

GGTCCCTGCG CAGGAGCCTG CGGCAGGCTG GGAATGCCGT TCTGGGAGAG CAGCCTCCCA

GGCGGGGTGG TGGGGAGACT GCAGGGAGGA GTTTTGGGGA GTGCAAACGG AAAGACTACA

TTTCCCAGGC CGCCACGCTT TCCAGCTGGA GTCCTAGGGC GCTGACTGCT CCCCAGTTTC

CGTAGGGAAG CGCTGGGCTA CCGCGGCTAT

(2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1079 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 1 (B) LOCATION: 1..634 (ix) FEATURE:
(A) NAME/KEY: exon 2 (B) LOCATION: 778..935 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 727..746 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 958..977 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 2:
TGTGCGTCGC GCTCTCGCCC TCCTCTTCCC GCTGCAGTGT CTATGGAGCG AGGCTACGTT
TCATTGCCGC CCTGGCTTAA CCCTTCCGGC GCCTAAAAGG ACGGCCGGCC GGCCGGTCCC

TTGCACCAGG AAGAAGTCTT AGCAGCCAGC GGGCCCTGGT CAGGAAACTC TAAGGTACAA

GGAAAACAGT TGAGGAAGGA GCCAGAGCGC TCCGGTTTGG TCCTCGGGCT TCGCTGGGGC

GGGGCGCAGG CGTTGGCTTT AAGAAAGGGG AGGGGACAGT GCAATCCGGG TTGCCCGCGG

ATTCGGCCAA GGAATCTTCC GCTCGCTCCG GAGCGAGGAG CCTGTAAAGA GGCTGTTCCC

AGCTCCAGCT CTAACCTCGC CTACACCTTG GGCGGGCCCA ATGTCACGTT TGCAATTGCT

CAGGAAGGAT CCGGCCCGTC TCCGGAGGCA AGTCGGGCTG CGGTTTTTGC TGCTTATCTG

GGAAGGCGAT GCCTAAGGGA CATGCTGCTT GCTAGGCAGC ACCCTGCCGG GATCCGACTG

CGATAGTTAG CTCTCCCTGG CCCTGAAGCC ATCGCCGGGG CGCCTGTTCT CTGTCCGGAC

CAGCCAGCGC TCCTCAGGAG TCTCACTGAA ACAGGTACCT GTCCTCCAAA GGGACGGAGG

CTATGAGCTT CCTTAAGCGG GTCGCGCGCT CAGTCCGTCC CCTCTACTTC CTCTACTGTG

CCATTGATGC TCTCGGTCTT TGTGTCTTTC CCCTTTCCCC CTACTCCCGG CCATCAGAAC

CATGGGTCAT CCCCCGCTGG AGTTCAGCGA CTGCTACCTG GACAGCCCCG ATTTCCGCGA

GAGGCTCAAG TGTTATGAGC AGGAACTGGA GAGGACCAAC AAATTCATCA AAGACGTAAT

CAAAGACGGC AACGCGCTTA TCAGCGCTAT GAGAAGTAAG TGCAAGGCTT CGATGAGCTG

TTTCTCTGAN CTGGTGTGTC TGGCCTTTAA GCCTTTCCAC ACCACCAGGG GAAGGGAGAT

TGCAGGGTGA CTCCCAGCCC ANATCTCTGA GGCAAATGGG TTTCCCACAC TTGGGGAGT

(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 900 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 3 (B> LOCATION: 403..498 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 375..394 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 504..523 ixi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:
CTTTGAACTC CACACGTATA AAATGGAAAG CATCTAGTGT ATTGCCCATA ATAGGGGTTT

AGGAAATGTG TAAGGAGACC CTCTTAAGAG TATATCTAAG TAGTTTCATG TCTTCCATGC

TTTGAGTGGA ACAGGTCAGA GAGGAGAGGT GTTGAGGATA AAAACRTGTC CCGTAACTTT

TAAGGACTTT ACTGAGATGC CCCATCCTTT CTTCTTGTGA TTTTAGTTGT TCAAACTTTC

TTCTACTGCA TATCTAATCT TTTGTTTTAT TTCATTAAAT GCTAGTTGCA ACCTGCTCAG

AGCACTTACC TTTGGTTTTC TTTTATCTGC TCTTATAGAG ATGAGGAAAT AGATCAAAGC

ATAGCATTGT TGAACATTTC TCTTGTTACC CTTCCTTTGC AGATTATTCT TCTGCTGTTC

AGAAATTTTC CCAGACGCTG CAGTCATTTC AGTTTGATTT CATTGGAGAC ACTCTGACTG

ATGATGAAAT TAACATCGGT AAGTCTTCAG CTACATGTGG TCATATACCT GTTGAGGCAG

CCCTGAGACC ATGTAGTCTT TTTGATTTGT GGATACAGAG CACTTGGACA TCTTCATCCA

CTGTGGTCCA ATGCCAAGGC CCTGGGAGGT TGATTAGGAA GGATCAGGAA ACTTTCCCTG

CCAGTCCCAT TTCCTCCTCA CACGACAGCA ATCAAAAGAT ACCCTTAAAC TTCTACTGAG

ATTTTTGACT CAGACAGTCT GCAAGCGACC TTTTCTTTAA AGCATAGTTA TTTTCCTAAA

GGATATATTA AAAGGGGGAC TTTTTTGGTT TATTTCCCAA AATGGTTGGA GTTAGATTCT

TCTAAGGAAT CAAATTTCCC TAGAAAGTGT TAAATTAGCA TTTGTGTGTC TACAACTTAT

(2) INFORMATION FOR SEQ ID N0: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 960 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 4 tB) LOCATION: 483..544 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 418..437 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 551..570 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 4:
TAGAAGAGTA GAAAGTTTGG GAGGGTGGAA AGGGTAATGT TTATTATTTA CTTTGTGTTG
ACACTACATT CACCATCCAA TTTAGTTTTT ACAGCAATAG TCTCTGGTTG GTCAGTAGTG

GAACAGGGAT CCAAACTCGT TTCAGGGTTC TATAGCTGCC AATTAATTAC ACAGCAAACC

TCTTGCCCCT TCCCTAATCT TTTAGCTTGT CTATGAAATA AGAGAAATTC TAGCTACTCC

TGAGTGGCTG TAAGGATTAA ATAAAATATT AAAGTGTATG GGGATTGATA AAAGAGGAAA

GAAAAGAAAA GAAACATTCA ACAGGTGCTG AACACCTGCT TTTGTCCTCC GATTTGACAC

CTTCCTCTAG TGGCCATGTG GGCATAGGGC ACTGGTCCCT ACTTCCTGTT GCACAGATCT

CTATCCATTT GTCTGTCAAG CACCATGATT AATTTGTTTT ACATTTGATT CTCTCCTTCC

AGCTGAATCC TTCAAGGAAT TTGCTGAATT GCTCAACGAG GTAGAAAATG AGAGGATGAT

GATGGTAAGT CACTAACGCT GTCACTGAAG CTGAGTTCAT GGGTGATATA GGGGATTTTT

CCTTTCCTTT ATGCTTGGAT TGATCCTATA CTATTTTGAT TTCTGTCAGA TAGCTTCTTG

GTGCTATAAA AATAGTTAGG TAATAGATCT GGTTATTATG TCTCAAGCTT CCACCCTGAG

AGTTTGGCAT TAGATAGAGG GAAATTAACG TGCAAATCCC ATCTGTGTTC ATTTCAGTGA

AAAATAATTT CAGTGGATAT TAAACTGGGC CTTTGAACAT GTTGACAGAA ATTGAGGTCT

TTAGTGTTTT TAGCCAAATT ATCCATTTGT TAATCTTTAA TTTGTGGAGT AGTTTTACTT

TTATAGAGAA AATCAGTAGA AAATAAAGAT AGAACTCATA TACCACCTTT CTCTCTCCCA

(2) INFORMATION FOR SEQ ID N0: 5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 960 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 5 (B) LOCATION: 451..522 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 423..445 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 553..574 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:
TCTGGTTTTA TGGCACAGGC AGGACTGTAA ATGTTTTATC CTAATGCTAT CACTGCACAC
TCATTTGCCT CTGATGRAAT GATTTGCACT TGCTGCAATT GTCCTTTTCT TGTATTTGCC

ATTCTCTTTC TCTTCTCTTT TTCACCTGTC CTTCAGGTCT CCTGTGTCCT CTGCTGGACA

TCACTTCAGA TATTTATCGA TTAAAAACTC AGGTCAGACT ATCATTAAAG TTACAGAGAA

ATGCCCCTCT TATTCTTTCT CCCATTTCTT CTCAATGCAT TTGATTTTTC AGAAACAATA

TAGAAACAAA CAGTAACAAA ACCCAACAAA TCAGCAAACC ATTTAACATT TTGCAGGTTG

GTATATAAAT GAAAATGTAG TAACAAGGAA TCTTGTATCT GAACCTTGTT AACCTAGAAA

TTGTTTTGTT TGTTTTTCCT TTTTGTCTAG GTACACAATG CTAGTGATTT GCTGATTAAA

CCCTTGGAAA ATTTCCGGAA GGAACAAATA GGCTTCACCA AGGTACATTT TCTGTATATG

CATAAGATTT TTTAAAATAG CAATCGAATA GTTGTATGGG CTACTATTCT TCACTTTACA

AAGATATGCA CCAATCTGCT GGTGCTTTGC TCTTGGCCTA GTCAGCCTCC TAAACTGTGC

AAAATAAATG TTTGTTGTTT ATGTCACCCT GTCTATGGCA TTCTGTTATA GTAGCCTCAG

CTAACATGAC AAAGGGGGTG GGGAGGTGGG TGATTAGTTT CTATGAGAAA ATGATCACGA

AAGAGAGTAA GAAAATCTAG AATTGGCCTC TGACTTTGTG GCCAACAGGC TCTGTATCTG

TGCATAAGTT TCTTCTTCTT TTGGTGTTTT TGTTGTTTTG TCTGGAAAAC TAGCTAGCTA

TCATGTATCA ACTGCCTGCT ATATTGAGCA CTAGGCTAGG TGCTTTACGT TCATTCTTTA

(2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1020 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 6 (B) LOCATION: 416..517 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 388..407 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 590..559 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:
CATTGTAGAT TAACCTTTTC ATGACAAATA TTTACTTCCA TTTGTGTTAT GTCTTCTTCC
TTTGTAP.GAA AATTTGAATG AGTGGTTAAT CTATGTGAAA AATATTGGAG GGAAGAAAAT

ATATCTACTG CACAGGCCCT TTTAAGGTAT CATTCTCTAA GGAGCAGCTT CCATAGCTTT

CAGCTGTAAA AATAGGGACT GCCATTTCTG CAGGCAGAAT GGTTTGGGGT TATATTTCAG

wo ~r~mo rc~r~Ew~siosss~

GAAGCTGAAA CTGCTGAGAC CAATACAAAG TTAATTCTCC GTTGCTTTTT TTTCCTTCCA

GGAGCTAGAG GGATTGACCA CCTGAAACCT GACACTATCT CCTTCATTCT CCTTTCTAAG

CAGACAGAGT TACAACTACA GACATTTAAT CTTTGCCCTT CTTTCCCACC TTTAGGAGCG

GAAAAAGAAA TTTGAAAAGG ATGGTGAGAG GTTTTATTCT TTACTGGATC GGCACTTACA

CCTGTCTTCA AAAAAGRAAG AATCTCAGTT ACAAGAGGTA TGTTCACAAA GCCTGCCCCT

GCCTTCCATT GCTAGCTATG CCTTAGAAAC AGTGTGAATT TTGTACTGCA AGGCTTTTCC

CATACCCCGT CTCAGCAGGG AACCTCATGT GATAGTAGCA CTTGTAGTCA AAACTGTGGC

CTGAGACTCA GAAGCCCTGA ATACTAAGCC AGCTCTTCCA CTAACTCAGG GTGTGACCTT

GGATAGGATA CTTCTTTCTT TGCCTCATTC CATTATCTGT AAGAAGAGGA GTCGAGAGTC

CCTTTCAATT GCAAGTCCAA AATCCATGCG AGGATAAAGT TAAACTAGTG TTGTATTTGG

TAGAAATCAG GAACAGATCT TCTACTTTTT TCCCTGAGAG ATTCCACAAC CTTTTTTTTT

TTTTTTTTTT TTGGTGAGGG GTTAGGGGAA TGTTCTGTTT TGGGACTACT GGTTACCTGG

GACTTGCAGT GCCTTCAGTT CAAATAAGCT ACCATTCGGT GAGACCCTAC CACATGCCAT

(2) INFORMATION FOR SEQ ID NO: ?:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1020 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 7 (B) LOCATION: 464..574 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 436..458 wo 99ma,~o rc°r~~srosss~
io (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 584..603 (xi) SEQUENCE DESCRIPTION; SEQ ID N0: 7:
CATTTCTTGT TAAATAAGAC TTGAGACTTG CAGTTCTTTC TGTTGTAATC CCCTCAGTAT
TGGCCATAGT AGCATGCTTC TGTTCTTGGG CTTTGGCCCT TGTTAATTTC TTATTCTCTG

TATTCATGTC TCTGTAGTGT TTGAGGGAGC AGATTTTCCT CTGACTTCAG TTCTCTGTTG

AATATAATAA AATGTGTGGA TTTTACACTT GTTCAGCTTT TTTCTTTTTG TCAGGGTGGA

TGTAATGACT TCAAGCTTTT TATGTGTTGG ACCAGAAACT GGAATCCTGT ATCAGTCACT

TTTTTATCTC TTGGCTTTAG GTTCTCTATT TCTAAAATAA AAGGTAACAT CACTAGTTGG

TGGCTAAATG CTCATCTAGC TCTAGCATTC TGTTCAATCC TTAAGTACTG ACTTTCATGA

ATGAATATCC CAATATGTAA TGTTTGTTTT CCTTCTTCTA CAGGCAGACC TACAGGTGGA

CAAGGAGAGG CACAATTTTT TCGAGTCCTC TCTTGATTAT GTTTATCAAA TCCAGGAAGT

TCAGGAGTCC AAGAAGTTCA ATATTGTGGA GCCTGTAAGT TTTCTCTGTT GATGAATGGT

CTAAAAP.TAT TTATCAAATG CCTGGTAAAT GTACAAACTT TGATCATAAA AACAGAATCC

CAAAGAACGT GAAGGATAAT CAGTGGAAAT GTCTAGAACG TAGTCTAACT TACGTTTTTC

TTGTTCTTTA GTATACTTTT TTTCTCTCTT TCCCCCCAAC CCTTTTCTCT CTTTTTCTAT

GGCTATTTTC TTTCTCTTTC CTTTTTTTCC TTTCTCTTTC CTCCCCCCTT CCGGAATTTC

TCTACAACTC TTCCTGTTAC TTTGTTTCCC TCCCTTCCTT CTTCTTGCGC TTTCAGAAGC

TTCTGAAAGT CTTTCTCATA TACCAGATAC TATGCTAGGG ACTCATGTCT CTTGCAGTCA

GTGACTTCTC ATTCTACTAC TTTTACTTCT GTCTTGTGTA GTTTTTTCCC ATTTCACGTT

wo 99r~mo pc~r~r9a~osss~
a (2) INFORMATION FOR SEQ ID N0: 8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 479 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 8 (B) LOCATION: 244..348 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 219..239 (ix) FEATURE:
(R) NAME/KEY: oligonucleotide (B) LOCATION: 363..381 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 8:
GCGCCACCAC GGCCAGCTAA TTTTTGTATT TTTAGTAGAG ATGGGATTTC ACCATGTTGG
TCAGGCTGGT CTCAAACTCC TGACCTCACG ATCTGCCCGC CTCGGCCTCC CAAAGTGCAG

GGATTACAGG CATAAGCCAC CACACCCAGC CACACCATGG AGTTTTTTGG TGAGTTCATG

TTTCTTTTAT TTAGTTTATT AGAAGATGCT GGTGATAAAG TTATTTTTAC ATGTTTTCCC

TAGGTCTTGG CCTTTCTTCA TAGTCTGTTC ATTTCTAACA GCCTGACTGT GGAGCTCACA

CAGGATTTCC TCCCATAGAA ACAACAGCTC CAACTCAGTT TACAGAATGT GAGTTTGCAT

GTGGATTTTT CTCACCGGTC TTTCCATTCC GATTGAATTT CAGCCCTAGC GACCTTGATT

CTTGGAATTC TAGGTTACTG CATCCTAGCC AATTTGTTAG AATATACTGG TGTGGATCA

f2) INFORMATION FOR SEQ ID N0: 9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 600~base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 9 (B1 LOCATION: 139..263 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 108..128 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 336..355 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:
ACCATTTCTT TTTTTGCTTT GTGGTTGCAC ATGCTGTAAG CAGGGAAAAC TTTGTACTGA
GTCTCTGACC AAGAAATACT TTTTCATGAT AATGATGATG ATAATAATGA TTTTCATGAT

GATGTCTTGA CAGACAAGAA ATCATTTCTC CAGTACCCGG GAAGAGATGG AAGAACTTAA

GAAAAGGATG AAAGAAGCTC CCCAGACATG CAAACTTCCA GGACAGCCAA CTATTGAAGG

CTATCTCTAT ACACAAGAGA AATGTGTGTG GGGACATAGG GGTATCCATT GGGTTTCAAT

AAGCCAGGAA GTAGTGCCAC TTGTCGGCTG TGAATTTTGG GCACCCTTAC TGTTCATAGA

CCCCTGATAG CTAAAATTCC CTTGGAACGC AGGCAGGGAA TACTGAAAAC p~~AAAAAApA

AAAGGAGAAA CTGAGAGGAA GTTAAAGATT TGTCTTACAA AGGCTGTGTA GTGATAAGAC

CTAAGGTTTT CTCTGAGATT CAAAATGGGT ATTATTTGTT CTTTAATCCT TCTGATTATT

CTTTTGGAAA AAAGGGAAGT AGAGGAAAGG AAGTAGAAAA ATAATATTTC TTATACTTAT

(2) INFORMATION FOR SEQ ID NO: 10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1020 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 10 (B) LOCATION: 383..483 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 361..380 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 992..511 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:
AGAGAGATGT GTAGAGTCAT GTACAGGTTA GAAGGGGCAT AACCCAATGA CCAATGACAT
GGAACAGTTA TAGAGACAGC AAAGTAAATG ACAATAGTCT CCAGCTTCTG GGATATATGG

TGATACAATT TATAGGGAAC ATGGCAGGGA GAGTAGGTTT TGAGCAAGCC AGAAGCAGCT

GTGGGAAGCA GTTGGTGAAG GTTAGAATCT CCCTGTGTGC AGTAGGTAGG TGGCTATGGA

AGGAGGGCGG TCAGGGCAAG GGCAGGGCTG GATCTGAAGT TTGACTCTGA AGAGCAATGT

GTAAATAGCT TCCATCTTAG GGTTGACTTC CTATACAGCT AAAATAGTTA TTCTGTCTGC

TCACTTTTAC TTGTCCTTGT AGGGGCTTTA GGRATATCCT GGGTGAAATA CTATTGCCAG

TATGAGAAAG AGACCRAAAC ACTGACCATG ACGCCTATGG AGCAGAAGCC AGGTGCTAAG

CAGGTCAGTT CTTGTTTGCA CCATATTTTT GGAAATGGAT CTATGACTGT TTCTCAGAAA

AGAATATATG TTGACCTAGT ATCAAATCAT CAAGTTCATC ACTGTTACGT GAGGCCATGA

CTTTATATGT ACACCTTGGC CTAAGTTTGA GTCAGATAGC ACTGAGTTGA GTGAAAAATT

TCTCTGTTGA TTAGAGCAAG CCTTTTGAAA GTGCCGGTAG TCTTTCAAAC CAGTTATTTT

TACAAGTGCC AGTCACATTG TACAGTCAAC TATGTAAAAA TATGGATGAA TTACTTTTAA

GAATGCTCTA CTCTTGGATT CTTTAAAATA GCAAGTTTTA AAAATATGAA TTGAATTCCA

AAATTCCTTT TTTACAGGAG TGTGTTTATG GCCCACAGTT GGAATAACCG ATACTCACAT

TCTATGTACT ACTCAAATAT CTTTAAGCAG TTAATCTCTC TTTTTCTGCC CTCCAAACCT

TCTCTCACTC CTGAAAATGA CAAGATAAAT TTAACACACT GAAAAAAATA GTTTACTTAC

(2) INFORMATION FOR SEQ ID NO: 11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 294 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear iii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 11 (B) LOCATION: 107..198 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 81..100 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 223..242 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:
CCCTAGACCA GACGCCTTCC AAGTGACACT GAAGACACTG TAGGAAGCAG AAACAGCTTC
TGTGTCTTTT TTTAAAAATT GCCTGTTAAT TCTTGTTTCA TATCAGGGGC CCTTGGACTT

AACACTGAAG TACTGTGTGA GAAGGAAGAC GGAGTCTATC GACAAGAGGT TCTGTTTTGA

CATAGAAACT AATGAAAGGT AAGCTGTGCC GCTGTGAATT GGCAATGTCC CCACGTGCCA

GATGCTTAGC CTGGGTATGT CTTTTATTTT CCTCCGTCAT CCCACGTTGA TGAC

(2) INFORMATION FOR SEQ ID NO: 12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 494 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double ZS
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 12 (B) LOCATION: 211..289 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 188..207 (ix) FEATURE:
(A} NAME/KEY: oligonucleotide (B) LOCATION: 300..319 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12:
CCACCCCCGC CTCCATGGTT CCCAAAGTTA CTGTTCTGTA AATTGCCTAT TGTTTTTCTT
GAGTAAAAGA GATTTTGAAA AATTAGTATT CTGTAAGCCT ATGCTTGTTC AGTGTGACAA

ACTCCAGGGC AGAGAGGGAC CCTAGATCAC AAGACTCCAT TCTCTCAGTT GAATTTTCTG

CTTTATACTT ACCATTTTTT TCCCCCTCAG GCCAGGAACC ATCACTCTGC AGGCCCTTTC

AGAAGCTAAC AGAAGGCTAT GGATGGAAGC CATGGATGGG AAAGAACCTG TAAGTTACCT

GACACTGGGG CAAACCTCCC CAGCATATGC CAGTGTATGA GTGCCCTCTA GTGGTATCAG

TGGGTCTCAN ACAATTAAAT GGTAATGGAT TGTTTAGTCT CAGTTTTAGA GCTGTAAGGR

ATTGTTTCCA CATCTCTTAG CAGGTAAGGC AACTGGAGTT CCAGAAAGGT TGAGGGACTT

TTCTGAGACC ACCC

(2) INFORMATION FOR SEQ ID NO: 13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 378 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) ivi) ORIGINAL SOURCE:

(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 13 (B) LOCATION: 212..245 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 166..189 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 259..278 (xi} SEQUENCE DESCRIPTION: SEQ ID NO: 13:
GGCATGAGCC ACCACGCCTG GCCTGTTCAA GTATTTTCTA GCAATCTTGG CAAAGCAATT
ATGTTTAGCC CACTTGGCTA TCTTTTTAAC ATCCTGGAGT TTCTAATCAT TTTTAATGCC

TATCTGGGGA AAGATATTTA ATATTATGTT CTCTGTTTTC CTATATTGAT TGACAATAGC

CATGGATCTT TCTGTTTATC TTCTTTTGTA GATCTACCAC AGCCCTATAA CAAAACAGCA

AGAAAGTGAG TCACTTAAGT TTTTGGTCTA CTAGCATTAT AAACTGCCAG CTGTCCGATT

CATAGTAAAT ACCATCATTA ATGATGTGTA CTACTAACGC AAGTCTGAAT ATGGATGCCT

TTGTGTGAAA TAAAATTC

(2) INFORMATION FOR SEQ ID N0: 14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 393 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 14 (8) LOCATION: 172..234 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 133..152 (ix) FEATURE:

(A) NAME/KEY: oligonucleotide (B) LOCATION: 250..269 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14:
AAAATGTTAA ACCTCCCCTG RAAAAATGAC TCTTTCCATT TAAGGGTGAC TAGAAATGAG
CAACTAAAAA CCCTTAGCTC TCTCAATGCA GTCCCTTTGC ATGGTCATTA AATGTTTAAT

AGGTGACACC TGTTGCAGCA GGATCTAACT CTTTTCCTTT GCTTGAAACA GTGGAGCTAA

ATGAAGTGGG CTTCAAGTTT GTCAGGAAGT GCATCAATAT TATTGAGACC AAAGGTAAGA

TCTGAACCAT AGTCTTGGCA TTGTCTGAAT CTCGTCACTC TGATTTTATC CTGGGCAATT

TCTCTGAAGT AGCGTTTTAG GAATGAAGAC TGTTTATAAA GCTTGTGTAG TAGATGCAAG

CTAGAAAATT TCAGAAAATT CTAAACTAGT GGT

(2) INFORMATION FOR SEQ ID NO: 15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 436 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 15 (B) LOCATION: 207..281 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 151..170 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 293..315 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:
AGATTAATGA GGGTTTGGTA CACTCCAAAT GAAAGGATGG ATAATTTGGA GAGATGCTGT
AGGACTATTC CCCTGTTACA GGGAAGGCTG GAGAACTTGG AGTATGTAGT GTGACCCCTT

WO 99/31230 PCf/EP98/08557 CCTATCTGAA TTGACTCTAG TGTACCAAGG GGAGATGACA ACTTTAGCTA TACAAGTGAA

ATTAACCTGA TTTTTTCCTC CACTAGGGAT CAAGACAGAA GGGTTGTACC GCACTGTGGG

CAGCAATATT CAGGTTCAGA AGCTGCTGAA TGCCTTTTTT GGTAACAATT TCACTTTGAT

AATTCTTATT GGGAGTACTT TATGTGTTAC AAAGAAATGT GACTGGAAGA GAAAGGAGAC

ACTGCTAAAA TGTGGTAGAA TAGTTGAAAA AAGTATTTTC TAAAGTAAAA CATACACATA

CTTGCCCACC CTGGGC

(2) INFORMATION FOR SEQ ID N0: 16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 54? base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL 50URCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 16 (B) LOCATION: 270..354 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 221..244 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 363..382 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16:
ACCCCAGTNA TNTGATGAAT CTAAGAAGAG TNGATNTTGT TTGTTCAGTT TTTTCTTGTT
GTGTGAATTG GATAGATTAC TTTNTTATTT CTTATATGGC AGACCAGAAT GCAGTCATGT

TTTTGAAATA TCAAAGATTT GCTTCTTCTA AAGTTTTGAT NTCTTAAAAA CTACTTAGGG

TNATATACTT TGTTTTTCTT TTAAAAGAGG GAAAATGTAA GATTTTTTTG ATGATTAACT

TTTGTTTTTT GTTTACTTTT CTCAAATAGA TCCTAAATGC CCAGGAGATG TTGATTTTCA

TAATAGTGAC TGGGACATTA AGACAATCAC CAGCTCCTTG AAATTCTACC TCAGGTATGC

CTGATTTGAA TTGGGAGTTT GCTTTTCATA GCTGGTGAAA TTTCTCTGGG TGTTGAGCGG

AGTTAACGTG GTCTCAGTTC CAGGAGTTTG GATACAATTG CTTAANAAAA AACATGTGAA

GAGGATTTCT GGCCANGAAT GTGCAAANAC TGTTTTTTAA ATCTGAGAGT TTAAGCAAGA

GAAGCAT

(2) INFORMATION FOR SEQ ID NO: 17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 601 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 17 (B) LOCATION: 355..413 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 305..324 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide IB) LOCATION: 438..457 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 17:
GGACAATTGC AAAAGCACTT CGGAAATTCT AAGGATCTAT CAAATCGTAA GGGATTCATG
GTAGCATTCA GCATGGGTCC CCTCTGGAAT TTTGCAGGAC TGGTTTGTGC CTCTTTTTAC

TTNTGGGAGC TAGTTGGAGA CCTTGCTAGA GGGCTCAGCC CATGCTTTTG CAGGTCTTTT

GTTGAATTAC TAGCAACTTG GATTCCCTGA CGAAGCTTCA GGTGAAGAGA AAAATGTATA

TAATCCCACT AAGCTGTAGG GCTCAGGAAC TTCAGCCTTG CTGTCCCCAG AACTAAGAAT

CCAATACCCA GCTGCTTTNT TCCCAAAGCA ACTGACAATT TTCATTCATT TCAGGAATCT

TTCTGAACCT GTCATGACCT ATAGACTTCA CAAAGAGCTG GTCTCTGCTG CCAGTAAGTA

TTTATGTTAC TAATTAACTG TGTTGTCCTA GTTTCTTAAT GTTTACTGCA ATAAGCCTAG

AAAATTGTTT GAGGGGAAGT GATTGAGGGC ACAGAAACCT AAAACACATA CACAAATTAT

GCACAACTGC CAAATGAAAG TATTCTTGCT TGCTGTCTAA CTCAANAATT CTATTATTTT

T

(2) INFORMATION FOR SEQ ID N0: 18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 387 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
.(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A} NAME/KEY: exon 18 (B) LOCATION: 80..185 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 25..44 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 218..237 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 18:
GTGCATATAT ATGTGAGAAT TTTGCTCAAT CCAGTAGCCC AGAAAGCCAA ACCATTTATC
TCTTACTGTT CTATCCCAGA GTCTGACAAC CTGGATTACC GCCTAGGAGC TATTCACTCC

CTGGTATATA AGCTACCAGA AAAGAACCGA GAGATGCTGG AACTTCTGAT AAGACACTTG

GTCAAGTAAG TAACTGCTGG ATTTTCAGAA AAAGTTCCTA TTAGAGGACT GGCCCATGTG

GTTGGACTAC ACAGAAACTG CCTCTCAGCT CTTTCAGCCC CAGCCCTTAA GTGCTTCCTT

GGAAGCTGAA TGCTCTGTGA GGAAGGCTAT TTTGCCTTGA CCCATGTACA TATCCTCTTA

GAGTCATCAT GCATGTGGAT TGTCTCA

(2) INFORMATION FOR SEQ ID NO: 19:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 460 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 19 (B) LOCATION: 79..238 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 51..70 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 252..271 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:
AACAGCACCT AAAACAGTCT TGGTTGTAAG GGGATACTGG AGCAAATTTT GTTAATCTTG
CCCCTTTTCT TCTGGCAGTG TGTGTGAGCA CAGCAAAGAG AATCTTATGA CCCCCTCCAA

CATGGGAGTA ATCTTTGGGC CCACCCTGAT GAGAGCTCAG GAGGACACTG TGGCCGCCAT

GATGAACATC AAATTCCAGA ACATAGTGGT GGAAATACTA ATCGAGCACT TTGGCAAGGT

ATGCATTTTC TATTCTCACT ACCTGTCTTC CAAACATGTG ACACTTTCCC CCAACTGCCT

TTTAGTGCTG TGTCTTCCTC CTTGGCTCAC GTTGACAGTG AAAGGAAATC CCATTATGAC

ACAATGACAT TTAATGGCAA CTCTGACCCT GGGAAATTCA TTCATTCAGC AAACATTGCT

TAAGCTTATA ACTATATTAT TTTCAGACAC CATGCTAAAT

WO 99/311.30 PCT/EP98/08557 (2) INFORMATION FOR SEQ ID NO: 20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 512 base pairs (B) TYPE: nucleic acid {C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A1 ORGANISM: Homo sapiens {ix) FEATURE:
(A) NAME/KEY: exon 20 (B) LOCATION: 230..377 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 206..225 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide {B) LOCATION: 383..402 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 20:
TCTTCCAAAC TGAGAAGTGC CAGGTTNTGT GCCTTGAGCA TAGTAGGAGN TACNTAAACA
TTTACCTGTA GNTAGAGTGA TTAAGAAAAT CTCTGATTCT TTGAGTCATG TTAGTATTCA

CGTNACAAAC TCTAGATATA AGGCCAACAA GCATCAANTG GTGGGTAGCA TTCAGAAGAC

AAAAANTTGA TNTAANTATT CTNTAGATAT NTTCCTTCTT TNTCCACAGA TCTATTTAGG

TCCACCTGAG GAAAGCGCTG CACCGCCAGT GCCTCCGCCT CGGGTGACAG CAAGAAGGCA

CAAACCAATC ACGATTTCAA AGCGCTTGCT GCGAGAAAGG ACGGTTTTCT ATACTTCTTC

CCTGGATGAA AGCGAAGGTC AGTACTNAGG TTCTCCTTTA GCTTCTGAAT GGTGATTAGA

CACNNAGNAN GATATCNAAT GGCTCAAGCG GTGGCATCAC CATTTNTCTC TCTATAAAAG

TANACCTTTC CTGNCTCCTG AACTTAAAAG CA

(2) INFORMATION FOR SEQ ID NO: 21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 841 base pairs WO 99/31230 PC1'/EP98108557 (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 21 (B) LOCATION: 185..508 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 151..170 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 511..530 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21:
GGCTTTACAT GAACTGGGAA GGGTAGAGAA TGATTTTGTG GGATATAGTT GGTTTGTGCC
ACAGTGACAT AACTGCTTTG AAAATGTATA CAAATTTTCA AAATTAAGTA TGTATGCATG

TATCAAAATG AAAAGGTTTT AAAAGTTATC ATTAATCTTC CCTNTTGGCA CCAACTTTTC

CTAGATGAAA TCCAACATCA AACACCGAAT GGTACTATCA CCAGCAGCAT AGAACCCCCC

AAGCCACCAC AACACCCCAA ACTACCTATT CAGAGGAGTG GGGAAACTGA TCCTGGGAGG

AAGTCCCCAA GCAGGCCTAT TTTGGATGGC AAGTTGGAGC CCTGCCCAGA GGTGGACGTG

GGGAAGTTGG TGTCTAGGCT GCAGGATGGA GGGACCAAGA TCACCCCAAA GGCCACCAAT

GGACCCATGC CAGGCTCTGG GCCCACCAAG ACCCCCTCTT TCCACATAAA GAGACCAGCT

CCCCGGCCCC TGGCCCACCA CAAGGAGGGT AAGTGCTTGG GAATCCCATG GGAGCCAGAG

CTGACCCTAA CTACTTTTCA CCTTGAGATC CTTCTGAGTT TGGAGATATA TTTAAGTGGA

AATATGTTCC AGTTTACTAC CACTAATATT GGAACAGTGG GCAAGATCAC AATAATCAGT

CACAATAATC ACTAGAATGT AAGCTCCATG AGGGCCGGGA TTTTTTACCT GTTTTGTTGA

CCTCTATATC CCAAGTGCTA TGTGCCTGGC ACTGTACTAA TTGCTGATAT ACTATTTCTT

ATCCTCACAA TCCCACTGTA AAGAATGTAT TATTCTTAAT ATTTTCTTTT TTTTTTTTTT
840 .
T

(2) INFORMATION FOR SEQ ID N0: 22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 615 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 22 (B) LOCATION: 320..485 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 294..313 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 496..515 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22:
TTTAATCCTC CCACTATCTC TGTAAGATAA TATATTGTGG ATCTTTATTA TATAGCTGGG
GAAACTGAGA CTTAGGGAAT GGATATGACA CACCCAAGAT ATNTGAAACT CCAGAGCTGG

GGTTCAAATA TAGACTTTCT GAAGGGACAG TTGCCAGAAA AATTACAAAA AAAAAi4AAAA

ATAGCCAGAG TTGTTAGTCA CCAAGAAGAA ATGGAGGCCA AGGAAGTTGG CCCAGGTAAC

TCTCATATTG GGTGCCTGCT CATGAGTAGT GTTCTGTTTG GCTAACCATC CAAGTTCCTG
300 ' GTATCATTTT CTCTTCCAGG GGATGCTGAC AGTTTCAGCA AAGTGCGGCC TCCAGGAGAA

AAGCCAACCA TCATCCGCCC CCCAGTGAGG CCCCCAGATC CTCCCTGCCG GGCAGCTACT

CCCCAAAAGC CAGAACCAAA GCCAGATATT GTGGCTGGCA ATGCGGGGGA AATCACATCA

TCTGTGTCAG TAGGGTTGTA CCTCAAAGTT GACTGAAGTC CTGTACTAGG CCACTAGGAA

TGCTTTCAGG ATCACCATAT TAAGGGTATA CAGTGCACAG CCCTGGGGCA TCCTTCACTT

TATAGTCTAG GGAAA

(2) INFORMATION FOR SEQ ID N0: 23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 475 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (ix) FEATURE:
(A) NAME/KEY: exon 23 (B) LOCATION: 211..261 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 179..198 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 271..291 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 23:
AATGGGGATA AAGAGTGTCA GCTATGGCCT TAGGGGTGCC TATGGGCTCT GGGCCATTTC
ACATTTGTAT GTGTAGGGCC TTTGCCAGCA AAGGCAGGGG CTGGCATTGG TGTCCCATCT

GGTTCAGAGT CTCCTGTCCT TTCTGTTGGC CATTGGTTCT CACGTGTATA CCAAAGCAAC

TTATGGGACT TGGTTGGCTT CTGTTTGCAG GGTGGCTTCC AGGACCAGGT TTTTTGAAAC

TTTAATAACT GCATCCTTAG CATACAATTG TGCTCACTCT AACATCTTTC TCTTTTTGTT

TCTCTACAGC TCTGTCTCTG TCTGTCACTT TCTCTTCCCC AATTCTGTCT CTCCATCCCT

ATCTGTCTGT CACCTGTTCA CCTGTGTGTC TATTTGTTTC TCTCATATTC TTTTT

(2) INFORMATION FOR SEQ ID NO: 24:
{i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 238 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: exon 24 (B) LOCATION: 115..156 {ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 65..84 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (8) LOCATION: 165..184 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 29:
CCATTTTTCA AATGGCCTCT TTAGCACTGG CCTAGAAGTG TCCCCCATTC CCCCAATTTA
CCTTTCCAGT CCTGATTTCT AGAATCTTAG TGAAACGTCT TTCTTTATCC ACAGTTCTCA

AGGCAGACTT CCTGGAGATG AAAGTTGAGG CTACAGGTAT GCAGTCCCCA TCCCTGATTA

CAAAATCTTG TTCCACATAA GCCTTCATTA CGGGATCTGA TATTTTGAGG ACTGGAAT

(2) INFORMATION FOR SEQ ID NO: 25:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4504 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
{A) NAME/KEY: exon 25 IB) LOCATION: 1..4235 (ix) FEATURE:
(A) NAME/KEY: polyA site (8) LOCATION: 4236. 4241 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25:
GTTTTAAAAG CCTTGGCCTC AGAGGACCCT TTCCAGGTTC TGAAAGAGTC TTTCTTCTTC
AAACCTTTGT GTGCGGAGTC ATTTTGTGTT GAAGAGCAGC TCCTTCCTAG CCTTGCACTT

TCAGACTCTC TCTGGGAGGC CATAAAATAA GGAGCATATG TCCTAGACAG GTGTTTATAT

CTCCTTTGTA TTCTGTCTTC ATCCCCTCAG AAGGTCTGTT TTGAGTTCCT ATAACACTGT

GAAGAGCTGG ACTCCCTCAA GCCAGACTCT GCCAAAACCA AGATATCCAC TTACCTGAGT

TGAAGAGGGG AGCTCAGTTT TCAACTCTTC CCTGAACTTC CTGCTTCCTC AGAGGGCCAT

TGAACTCTGA GAGATTTGGG GCTAAAGACT GATCTCAGGG GTCTTACCTT GAACTGAAGG

CCACTTGAGT TGGGGCCATT GCTTACCTTG GTTGGAAGGG AATAGAAATG TTTGCTGAAC

ATTGGAGAAT CTCAACATGT CTCCTACTGA GGATATGGAC ACTGGTGCCA TGTCAGCGCT

CTGGTGCTGC AGTATGTTGC CAAGAGCCCG TCTGCTCCTG CGAGGCTATG AGTGGGATGA

GTGATGCCCC CACAGCACCT CCATGTGGAC TTAGGAAGGT GGCCTTCCTG CTGTTACATG

CAGCCACTTA GGACAAATCT GCAAAGCATG TTTTGCATGT AAAAGCCTAG GTCTATTTGG

ATTATTCTTT CTCGTTTTTT TTGACAGCTT CCTGTCAAGC AATCAAGAAA CAAACAAAAG

CTGAACACAT TTCCTTTTAA AAAAAGGAGA CTGTGTTTGG TCCTGTAGGA GTTCTATTTT

GGGGTCAAAT GCTAGAAAAA TTGTTAAGGT GGATTGAGGC CAGGCAGCTG TCACTGCTGC

TTCATGTTTG CCTTCTGCAC ATAAACTCTT TTATCTCCTG AAAAAAGCAG TTCTTAACCC

AGTGTCCATG GACTCAGAAA CTCCATGATG CCCCTGAGAT GGTATGCACA ATTCCATGAC

AATATGCCCT TTCTGGGGAG ATAGTCCATA ATGTTCTGCT AAATTTCAAA TGGGCTCGTG

ACCCAAAAAA GTCAAGAACC ACAGCACTTG AGTTAAAATA CTCTTTTTAC AATCCATATA

AGCCCTTGAT TGGAAGGGCT TTTCAAAATC ATTTAGTCTA ACAACTGCCC AGTTTCCAGT

CGGGGGRACT GAGGCAGAGC AAGGTAGTGA TCACACCAGT ACAAGATTTC AGGTCCCAGG

CTCCTATGCA AGTTTTTTTT CCCCATTATA TCACACTTAT TTAGCAAGGG ACCTTGTGGT

TTGTGGCTTT AGTGGCCATC ATTTCTGGGG GTTGGCTTTT ACCCTTTTTC TTGAATATTT

GCCACCAAGT GAAAAATGTT AGGACATAAA CCCTTGCCAG GTCCCTTTCA TTTGCTATCT

CTATTATTGG AAAGGACCTA AAAATTGGTG TAATGGGGCA GAAATCTGAG GAATGGACAT

TTCTAATTCC TGTTTGTTGA AGGGAAGTTG CTGGAAAGAG CATCAGTACT TGTTTCTATG

CAGATGCCTG GGCCGTAGCT TGTCTGTAGC GTCTGTATAA TTATAATGTT GCCCAGTGTG

AGGGAAAGAG CTTTCCTACT TGCACTCTTC TACCAAGGCC CTGTTAGTGC ACTGATTATA

GTACTGACAG ATAAAGCCTA GATGAGAGAG ATAGAGAGTG AGTACATGCA CACTCATGTG

CAAACCCACT CAGAGATGCA TTTGGAACAG TGCTACTGAA AGGCAGTAGT CATTTTCAAG

ACTGAATTCC AAACATGGTT TATTGGTGAG TTAGGAACAT GTAAGGCCAA GTACACTGAG

AGCCTTTTTG AAAGTAATTG AGTGGAAACT TGATGCCATT CTAAATCAAG GCATATCCAG

GTGGCCCGGT TTGAACTCAC TCCACTGTAC CCAGTCTCAA AGGCCAGGTT GCTAAGAAAC

CAGGAGTAAA AGAGTCAAGT GACCATCATT TCACCTGCTG CTTGCCCCCA ATAGTAGTCT

CTGTGAGGCC TTACTGACCT CACCTAGGAA GTGATTTTTG AGCCCTTGTT TCAGGGCTGT

GGCCTCCCTG CTCTATCCTG AATAAAGCAG ACAGGTGTGC AGATTTTGGC CATGAAAGCA

TGGCTAATAG GGCCACAGTC CCTTTAAAGA AACATGGTTT GACTCTGGTT TTCTTGGGGG

AAAATACCAC AATCACCGAT GCAAACATTG GAAGATTATT GAGAGCCCTA GAAAGCTGCT

GTGATCCCAG TAGAAAATAT GTCCCAGAAA TGTCATGAGA TTGCTGTGTG TTGCCTGGGA

CACAGATCAA GGGCCTATCT TGGAGAGCTG GGGACCAGCA GTCTGCCTGG AGGCCAGGGA

GCAGTGGCTG AGTAGCTCTG CCTTTGCTCT GGTCTATACC TTAAGAATGC CAAAGAATGA

ATTTCAACGC CTGCCTTTGG CACTCTGACT TAAAGTGCAA AAAGCTTCTG TGGCGAGGCA

TGCTATCATG GAATGAGACT GGCTTGCCCT AGGCTTAATG GATGGGCAGT CATTTTGCAG

AGGCTATGGG AAGAGGGTGA TAATAGAAGA GTGGCAGCTA TAGGAAATTA TCAACATACC

TTGGCCAGCA AGTTAGAGAA TCTGGCAATG GATGAACTGA AAGTGATGAA CTGGCAGGGA

TAACAAAGAA CCTAACATTT ATTAAGCACG TATTTATTAA CTGCTCAGTG TTTCATATTC

ATGCAAGTAT TCTCATTTTA CAGAGAAAGA AATTATGGCC CAGGGGGCTA AAGTAAACAA

CTCAAGGGCA CATAGAAAGT AAATAAAAGG ACTGTGATTT GAATCCAGGC CACTCTTAGC

CCATGCTGTT TTCCCTTTGC CAGACTGTGG TAGGTGTTTG AACAGAGGCC ACATTACTAG

AGTTGGCATG ACTCTTGACT CTTGCCTGCC TAACAAAATA TTGAAAGGCA AACATTTGAA

GGAGGGAGGG GGTGCAGGTT CAGTTTATAT GGAAATGCAA ACTGGGCTGG AAGATATTCC

TGAGTTAGGC TTTCTCTTCA TATTCAGCTT GCACATTTGG TAATGTTTTT AAAATGATCA

TCTAATTTTA TTTTGTGAAG TGAAGGATTT GTGTTTTAGT TGGCAGTTGT TAAGTCCTTG

GCTTGCCATT TTTCAAAAAG TAAAAAGGTC CTCACAGGTG TCTCCATACT TCGCCAAGGT

TGTAGCATGG GCAGTTTCAG TTTCAGCCTA AGAGACTGGT GACATCCACA AATGCAGTTT

TAGAAGCAGA AAAGGTCTTG GTGCCTCTGC AGTACTTGAT GTATTGGGGT CAAATCTCTA

CAAATTTTTC TGTGGTGATA GCAAAATCAA GAGATGGCTT ACAAAAAGAA ATATTGAATT

TTTATTTTTG AAGTTTTTGT TTTTTAAAAG GTTGGGGGTG TTCAGCCACT GAGGGACAAA

ACTTAGCATC TAATTTCAAT TATAGTGTCA TGCAGAGTAT TTCTAAAGTA ATTGGTTATC

ATGGGAAAGT ATTCTCTTTT CAAGAAGTTC TTTGATTCTG TAATAACTAG AACAAATAAA

GTAGTAAAAG AAGAAATAGT TCTGTGACTA GGAAAAAATT GCTTTTGAGA GAACATAGAT

CAATTATACT ACTTCTAAGG TAGCTGCAGA TAAGTGGCCT TGACACATTA CAAGCCTGGA

AAAAAACATC AGAAATAATA AAAAP.TTTCA GAGAGAATCA AGATACCTTT TTTTTTCTTT

TTTTTTTCTT TTTTTTATTA TACTCTAAGT TTTAGGGTAC ATGTGCACAT TGTGCAGGTT

AGTTACATAT GTATACATGT GCCATGCTGG TGCGCTGCAC CCACTAATGT GTCATCTAGC

ATTAGGTATA TCTCCCAGTG CTATCCCTCC CCCCTCCCCC GACCCCACCA CAGTCCCCAG

AGTGTGATAT TCCCCTTCCT GTGTCCATGT GATCTCATTG TTCAATTCCC ACCTATGAGT

GAGAATATGC GGTGTTTGGT TTTTTGTTCT TGCGATAGTT TACTGAGAAT GATGGTTTCC

AATTTCATCC ATGTCCCTAC AAAGGATATG AACTCATCAT TTTTTATGGC TGCATAGTAT

TCCATGGTGT ATATGTGCCA CATTTTCTTA ATCCAGTCTA TCATTGTTGG ACATTTGGGT

TGGTTCCAAG TCTTTGCTAT TGTGAATAGT GCCGCAATAA ACATACGTGT GCATGTGTCT

TTATAGCAGC ATGATTTATA CTCATTTGGG TATATACCCA GTAATGGGAT GGCTGGGTCA

AATGGTATTT CTAGTTCTAG ATCCGTGAGG AATCGCCACA CTGACTTCCA CAATGGTTGA

ACTAGTTTAC AGTCCAACCA ACAGTGTAAA AGTGTTCCTA TTTCTCCGCA TCCTCTCCAG

CACCTGTTGT TTCCTGACTT TTTAATGATT GCCATTCTAA CTGGTGTTGA GATGATATCT

CATA

i2) INFORMATION FOR SEQ ID NO: 26:

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3101 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo Sapiens (ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 639..3047 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 152..1?2 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 586..606 (ix) FEATURE:
(A) NAME/KEY: oligonucleotide (B) LOCATION: 641..663 (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 26:
TGTGCGTCGC GCTCTCGCCC TCCTCTTCCC GCTGCAGTGT CTATGGAGCG AGGCTACGTT
60 ~ .
TCATTGCCGC CCTGGCTTAA CCCTTCCGGC GCCTAAAAGG ACGGCCGGCC GGCCGGTCCC

TTGCACCAGG AAGAAGTCTT AGCAGCCAGC GGGCCCTGGT CAGGAAACTC TAAGGTACAA

GGAAAACAGT TGAGGAAGGA GCCAGAGCGC TCCGGTTTGG TCCTCGGGCT TCGCTGGGGC

GGGGCGCAGG CGTTGGCTTT AAGA,AAGGGG AGGGGACAGT GCAATCCGGG TTGCGCGCGG

ATTCGGCCA,A GGAATCTTCC GCTCGCTCCG GAGCGAGGAG CCTGTAAAGA GGCTGTTCCC

AGCTCCAGCT CTAACCTCGC CTACACCTTG GGCGGGCCCA ATGTCACGTT TGCAATTGCT

CAGGAAGGAT CCGGCCCGTC TCCGGAGGCA AGTCGGGCTG CGGTTTTTGC TGCTTATCTG

GGAAGGCGAT GCCTAAGGGA CATGCTGCTT GCTAGGCAGC ACCCTGCCGG GATCCGACTG

CGATAGTTAG CTCTCCCTGG CCCTGAAGCC ATCGCCGGGG CGCCTGTTCT CTGTCCGGAC

CAGCCAGCGC TCCTCAGGAG TCTCACTGAA ACAGAACCAT GGGTCATCCC CCGCTGGAGT

TCAGCGACTG CTACCTGGAC AGCCCCGATT TCCGCGAGAG GCTCAAGTGT TATGAGCAGG

AACTGGAGAG GACCAACAAA TTCATCAAAG ACGTAATCAA AGACGGCAAC GCGCTTATCA

GCGCTATGAG AAATTATTCT TCTGCTGTTC AGAAATTTTC CCAGACGCTG CAGTCATTTC

AGTTTGATTT CATTGGAGAC ACTCTGACTG ATGATGAAAT TAACATCGCT GAATCCTTCA

AGGAATTTGC TGAATTGCTC AACGAGGTAG AAAATGAGAG GATGATGATG GTACACAATG

CTAGTGATTT GCTGATTAAA CCCTTGGAAA ATTTCCGGAA GGAACAAATA GGCTTCACCA

AGGAGCGGAA AAAGAAATTT GAAAAGGATG GTGAGAGGTT TTATTCTTTA CTGGATCGGC
loeo ACTTACACCT GTCTTCAAAA AAGAAAGAAT CTCAGTTACA AGAGGCAGAC CTACAGGTGG

ACA,AGGAGAG GCACAATTTT TTCGAGTCCT CTCTTGATTA TGTTTATCAA ATCCAGGAAG
lzoo TTCAGGAGTC CAAGAAGTTC AATATTGTGG AGCCTGTCTT GGCCTTTCTT CATAGTCTGT

TCATTTCTAA CAGCCTGACT GTGGAGCTCA CACAGGATTT CCTCCCATAC AAACAACAGC

TCCAACTCAG TTTACAGAAT ACAAGAAATC ATTTCTCCAG TACCCGGGAA GAGATGGAAG

AACTTAAGAA AAGGATGAAA GAAGCTCCCC AGACATGCAA ACTTCCAGGA CAGCCAACTA

TTGAAGGCTA TCTCTATACA CAAGAGAAAT GGGCTTTAGG AATATCCTGG GTGAAATACT

ATTGCCAGTA TGAGAAAGAG ACCAAAACAC TGACCATGAC GCCTATGGAG CAGAAGCCAG

GTGCTAAGCA GGGGCCCTTG GACTTAACAC TGAAGTACTG TGTGAGAAGG AAGACGGAGT

CTATCGACAA GAGGTTCTGT TTTGACATAG AAACTAATGA AAGGCCAGGA ACCATCACTC

TGCAGGCCCT TTCAGAAGCT AACAGAAGGC TATGGATGGA AGCCATGGAT GGGAAAGAAC

CTATCTACCA CAGCCCTATA ACAAAACAGC AAGApp,TGGA GCTAAATGAA GTGGGCTTCA

AGTTTGTCAG GAAGTGCATC AATATTATTG AGACCAAAGG GATCAAGACA GAAGGGTTGT

ACCGCACTGT GGGCAGCAAT ATTCAGGTTC AGAAGCTGCT GAATGCCTTT TTTGATCCTA

AATGCCCAGG AGATGTTGAT TTTCATAATA GTGACTGGGA CATTAAGACA ATCACCAGCT

CCTTGAAATT CTACCTCAGG AATCTTTCTG AACCTGTCAT GACCTATAGA CTTCACAAAG

AGCTGGTCTC TGCTGCCAAG TCTGACAACC TGGATTACCG CCTAGGAGCT ATTCACTCCC

TGGTATATAA GCTACCAGAA AAGAACCGAG AGATGCTGGA ACTTCTGATA AGACACTTGG

TCAATGTGTG TGAGCACAGC AAAGAGAATC TTATGACCCC CTCCAACATG GGAGTAATCT

TTGGGCCCAC CCTGATGAGA GCTCAGGAGG ACACTGTGGC CGCCATGATG AACATCAAAT

TCCAGAACAT AGTGGTGGAA ATACTAATCG AGCACTTTGG CAAGATCTAT TTAGGTCCAC

CTGAGGAAAG CGCTGCACCG CCAGTGCCTC CGCCTCGGGT GACAGCAAGA AGGCACAAAC

CAATCACGAT TTCAAAGCGC TTGCTGCGAG AAAGGACGGT TTTCTATACT TCTTCCCTGG

ATGAAAGCGA AGATGAAATC CAACATCAAA CACCGAATGG TACTATCACC AGCAGCATAG

AACCCCCCAA GCCACCACAA CACCCCAAAC TACCTATTCA GAGGAGTGGG GAAACTGATC

CTGGGAGGAA GTCCCCAAGC AGGCCTATTT TGGATGGCAA GTTGGAGCCC TGCCCAGAGG

TGGACGTGGG GAAGTTGGTG TCTAGGCTGC AGGATGGAGG GACCAAGATC ACCCCAAAGG

CCACCAATGG ACCCATGCCA GGCTCTGGGC CCACCAAGAC CCCCTCTTTC CACATAAAGA

GACCAGCTCC CCGGCCCCTG GCCCACCACA AGGAGGGGGA TGCTGACAGT TTCAGCAAAG

TGCGGCCTCC AGGAGAAAAG CCAACCATCA TCCGCCCCCC AGTGAGGCCC CCAGATCCTC

CCTGCCGGGC AGCTACTCCC CAAAAGCCAG AACCAAAGCC AGATATTGTG GCTGGCAATG

CGGGGGAAAT CACATCATCT GTGGTGGCTT CCAGGACCAG GTTTTTTGAA ACAGCTTCCC

GGAAAACAGG AAGTTCTCAA GGC.1GACTTC CTGGAGATGA AAGTTGAGGC TACAGGTTTT

AAAAGCCTTG GCCTCAGAGG ACCCTTTCGA GGTTCTGAAA G

(2) INFORMATION FOR SEQ ID N0: 27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 802 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (vi) ORIGINAL SOURCE:
(A) ORGANISM: Homo sapiens (xi) SEQUENCE DESCRIPTION: SEQ ID N0: 27:
Met Gly His Pro Pro Leu Glu Phe Ser Asp Cys Tyr Leu Asp Ser Pro Asp Phe Arg Glu Arg Leu Lys Cys Tyr Glu Gln Glu Leu Glu Arg Thr Asn Lys Phe Ile Lys Asp Val Ile Lys Asp Gly Asn Ala Leu Ile Ser Ala Met Arg Asn Tyr Ser Ser Ala Val Gln Lys Phe Ser Gln Thr Leu Gln Ser Phe Gln Phe Asp Phe Ile Gly Asp Thr~Leu Thr Asp Asp Glu Ile Asn Ile Ala Glu Ser Phe Lys Glu Phe Ala Glu Leu Leu Asn Glu Val Glu Asn Glu Arg Met Met Met Val His Asn Ala Ser Asp Leu Leu Ile Lys Pro Leu Glu Asn Phe Arg Lys Glu Gln Ile Gly Phe Thr Lys Glu Arg Lys Lys Lys Phe Glu Lys Asp Gly Glu Arg Phe Tyr Ser Leu Leu Asp Arg His Leu His Leu Ser Ser Lys Lys Lys Glu Ser Gln Leu Gln Glu Ala Asp Leu Gln Val Asp Lys Glu Arg His Asn Phe Phe Glu ser Ser Leu Asp Tyr Val Tyr Gln Ile Gln Glu Val Gln Glu Ser Lys WO 99!31230 PCT/EP98/08557 Lys Phe Asn Ile Val Glu Pro Val Leu Ala Phe Leu His Ser Leu Phe Ile Ser Asn Ser Leu Thr Val Glu Leu Thr Gln Asp Phe Leu Pro Tyr Lys Gln Gln Leu Gln Leu Ser Leu Gln Asn Thr Arg Asn His Phe Ser Ser Thr Arg Glu Glu Met Glu Glu Leu Lys Lys Arg Met Lys Glu Ala Pro Gln Thr Cys Lys Leu Pro Gly Gln Pro Thr Ile Glu Gly Tyr Leu Tyr Thr Gln Glu Lys Trp Ala Leu Gly Ile Ser Trp Val Lys Tyr Tyr Cys Gln Tyr Glu Lys Glu Thr Lys Thr Leu Thr Met Thr Pro Met Glu Gln Lys Pro Gly Ala Lys Gln Gly Pro Leu Asp Leu Thr Leu Lys Tyr Cys Val Arg Arg Lys Thr Glu Ser Ile Asp Lys Arg Phe Cys Phe Asp Ile Glu Thr Asn Glu Arg Pro Gly Thr Ile Thr Leu Gln Ala Leu Ser Glu Ala Asn Arg Arg Leu Trp Met Glu Ala Met Asp Gly Lys Glu Pro Ile Tyr His Ser Pro Ile Thr Lys Gln Gln Glu Met Glu Leu Asn Glu Val Gly Phe Lys Phe Val Arg Lys Cys Ile Asn Ile Ile Glu Thr Lys Gly Ile Lys Thr Glu Gly Leu Tyr Arg Thr Val Gly Ser Asn Ile Gln ValGln LysLeu Leu PhePheAsp ProLysCys ProGly Asn Asp Rla ValAsp PheHis AsnSer TrpAspIle LysThrIle ThrSer Ser Asp LeuLys PheTyr LeuArg LeuSerGlu ProValMet ThrTyr Arg Asn LeuHis LysGlu LeuVal AlaAlaLys SerAspAsn LeuAs T
Ser p yr ArgLeu GlyAla IleHis LeuValTyr LysLeuPro Gl Ser u Lys Asn ArgGlu MetLeu GluLeu IleArgHis LeuV
Leu l a Asn ValCys Glu His Ser Lys Glu Asn Leu Met Thr Pro Ser Asn Met Gly Val Ile Phe Gly Pro Thr Leu Met Arg Ala Gln Glu Asp Thr Val Ala Ala Met Met Asn Ile Lys Phe Gln Asn Ile Val Val Glu Ile Leu Ile Glu His Phe Gly Lys Ile Tyr Leu Gly Pro Pro Glu Glu Ser Ala Ala Pro Pro Val Pro Pro Pro Arg Val Thr Ala Arg Arg His Lys Pro Ile Thr Ile Ser Lys Arg Leu Leu Arg Glu Arg Thr Val Phe Tyr Thr Ser Ser Leu Asp Glu Ser Glu Asp Glu Ile Gln His Gln Thr Pro Asn Gly Thr Ile Thr Ser Ser Ile Glu Pro Pro Lys Pro Pro Gln His Pro Lys Leu Pro Ile Gln Arg Set Gly Glu Thr Asp Pro Gly Arg Lys Ser Pro Ser Arg Pro Ile Leu Asp Gly Lys Leu Glu Pro Cys Pro Glu Val Asp Val Gly Lys Leu Val Ser Arg Leu Gln Asp Gly Gly Thr Lys Ile 'ithr Pro Lys Ala Thr Asn Gly Pro Met Pro Gly Ser Gly Pro Thr Lys Thr Pro Ser Phe His Ile Lys Arg Pro Ala Pro Arg Pro Leu Ala His His Lys Glu Gly Asp Ala Asp Ser Phe Ser Lys Val Arg Pro Pro Gly Glu Lys Pro Thr Ile Ile Arg Pro Pro Val Arg Pro Pro Asp Pro Pro Cys Arg Ala Ala Thr Pro Gln Lys Pro Glu Pro Lys Pro Asp Ile Val Ala Gly Asn Ala Gly Glu Ile Thr Ser Ser Val Val Ala Ser Arg Thr Arg Phe Phe Glu Thr Ala Ser Arg Lys Thr Gly Ser Ser Gln Gly Arg Leu Pro Gly Asp eoo Glu Ser

Claims (25)

31

1. Nucleic acid, the sequence of which is selected from the group consisting of sequences SEQ ID n°1 to SEQ ID n° 25, and a homologous nucleic acid sequence thereof.

2. Nucleic acid, the sequence of which is selected from the group consisting of exon sequences as identified in table 2, and a homologous nucleic acid sequence thereof,

3. Nucleic acid comprising the sequence as shown in SEQ ID n°
26, a homologous sequence thereof, or a sequence identical to SEQ ID n°
26.
except for a one base deletion of the nucleotide 1578 as shown in SEQ ID
N° 26.

4. isolated oligophrenin 9 polypeptide substantially comprising the aminoacid sequence of SEQ ID n° 27, or a homologous amino acid sequence thereof.

5. Vector for cloning and/or expression comprising a nucleic acid sequence of any of Claims 1 to 3.

6. Host cell transfected with a vector according to claim 5.

7. Nucleic acid, the sequence of which has at feast 15 bases and specifically hybridizes with a nucleic acid sequence according to any of claims 1 to 2, under stringent conditions.

8. Nucleic acid, the sequence of which has at least 15 bases and specifically hybridizes with a nucleic acid sequence according to claim 3 under stringent conditions.

9. Nucleic acid of claim 7 or 8, the sequence of which is selected from the group consisting of the sequences identified in table 3 or the complementary sequences thereof, said sequences identified in table 3 consisting of:
- nucleotide n° 727 to 746 of SEQ ID n° 2 - nucleotide n°958 to 977 of SEQ ID n°2 - nucleotide n°375 to 394 of SEQ ID n°3 - nucleotide n°504 to 523 of SEQ ID n°3 - nucleotide n°418 to 437 of SEQ ID n°4 - nucleotide n°551 to 570 of SEQ ID n°4 - nucleotide n°423 to 445 of SEQ ID n°5 - nucleotide n°553 to 574 of SEQ ID n°5 - nucleotide n°388 to 407 of SEQ ID n°6 - nucleotide n°540 to 559 of SEQ ID n°6 - nucleotide n°436 to 458 of SEQ ID n°7 - nucleotide n°584 to 603 of SEQ ID n°7 - nucleotide n°219 to 239 of SEQ ID n°8 - nucleotide n°363 to 381 of SEQ ID n°8 - nucleotide n°108 to 128 of SEQ ID n°9 - nucleotide n°336 to 355 of SEQ ID n°9 - nucleotide n°361 to 380 of SEQ ID n°10 - nucleotide n°492 to 511 of SEQ ID n°10 - nucleotide n°81 to 100 of SEQ ID n°11 - nucleotide n°223 to 242 of SEQ ID n°11 - nucleotide n°188 to 207 of SEQ ID n°12 - nucleotide n°300 to 319 of SEQ ID n°12 - nucleotide n°166 to 189 of SEQ ID n°13 - nucleotide n°259 to 278 of SEQ ID n°13 - nucleotide n°133 to 152 of SEQ ID n°14 - nucleotide n°260 to 269 of SEQ ID n°14 - nucleotide n°151 to 170 of SEQ ID n°15 - nucleotide n°293 to 315 of SEQ ID n°15 - nucleotide n°221 to 244 of SEQ ID n°16 - nucleotide n°363 to 382 of SEQ ID n°16 - nucleotide n°305 to 324 of SEQ ID n°17 - nucleotide n°438 to 457 of SEQ ID n°17 - nucleotide n°25 to 44 of SEQ ID n°18 - nucleotide n°218 to 237 of SEQ ID n°18 - nucleotide n°51 to 70 of SEQ ID n°19 - nucleotide no 252 to 271 of SEQ ID no 19 - nucleotide no 206 to 225 of SEQ ID no 20 - nucleotide no 383 to 402 of SEQ ID no 20 - nucleotide no 151 to 170 of SEQ ID no 21 - nucleotide no 511 to 530 of SEQ ID no 21 - nucleotide no 294 to 313 of SEQ ID no 22 - nucleotide no 496 to 515 of SEQ ID no 22 - nucleotide no 179 to 198 of SEQ ID no 23 - nucleotide no 271 to 291 of SEQ ID no 23 - nucleotide no 65 to 84 of SEQ ID no 24 - nucleotide no 165 to 184 of SEQ ID no 24 - nucleotide no 152 to 172 of SEQ ID no 26 - nucleotide no 586 to 606 of SEQ ID no 26 - nucleotide no 641 to 663 of SEQ ID no 26

10. Method for producing a recombinant oligophrenin 1 polypeptide, wherein a host cell of claim 6 is cultured in conditions allowing the expression of a polypeptide according to claim 4.

11. Monoclonal or polyclonal antibodies, or fragments thereof.
chimeric or immunoconjugate antibobies, which are capable of specifically recognizing a polypeptide according to claim 4.

12. Use of the antibodies of claim 11 for detecting or purifying a polypeptide according to claim 4 in a biological sample.

13. Use of a nucleic acid according to any of claims 1, 2, 3, 7, 8 and 9, for detecting an abnormality in the oligophrenin 1 gene or in the transcripts of the oligophrenin gene.

14. Method of in vitro diagnosis of a neurological disorder associated with an abnormality in the oligophrenin 1 gene or in the transcripts of the oligophrenin 1 gene, wherein one or more mutation(s) is detected in any of the sequences of claims 1 to 3.

15. A method according to claim 14 wherein said mutation is a one base deletion of the nucleotide 1578 as shown in SEQ ID no 26.

16. Method of in vitro diagnosis according to any of claims 14 or 15 comprising the steps of:
- contacting a biological sample containing DNA with specific oligonucleotides having a sequence as defined in claim 7, permitting the amplification of all or part of the oligophrenin 1 gene, the DNA contained in the sample having being rendered accessible, where appropriate, to hyrbridization, and under conditions permitting a hybridization of the oligonucleotides with the DNA
contained in the biological sample ;
amplifying said DNA ;
- detecting the amplification products ;
- comparing the amplified products as obtained to the amplified products obtained with a normal control biological sample, and thereby detecting a possible abnormality in the oligophrenin 1 gene.

17. Method of in vitro diagnosis according to any cf claims 14 or 15 comprising the steps of:
- producing cDNA from mRNA contained in a biological sample ;
- contacting said cDNA with specific oligonucleotides haying a sequence as defined in claim 8, permitting the amplification of all or part of the transcript of the oligophrenin 1 gene, under conditions permitting a hybridization of the primers with said cDNA ;
- amplifying said cDNA ;
- detecting the amplification products ;
- comparing the amplified products as obtained to the amplified products obtained with a normal control biological sample, and thereby detecting a possible abnormality in the transcript of the oligophrenin 1 gene.

18. Pharmaceutical composition comprising a purified oligophrenin 1 polypeptide of claim 4 and/or a homologous polypeptide thereof, or an isolated nucleic acid sequence encoding said polypeptide in association with a pharmaceutically acceptable carrier

19. Pharmaceutical composition comprising an anti-sense sequence as defined in claim 7 or 8 in association with a pharmaceutically acceptable carrier.

20. Pharmaceutical imposition comprising an antibody according to claim 11.

21. Transgenic non-human mammal expressing an exogenous oligophrenin 1 protein as defined in claim 4, or being modified so as to overexpress a native oligophrenin 1 protein as defined in claim 4, or so as to express a non-functional oligophrenin 1 protein as defined in claim 4.

22. Method for screening drugs likely to act on the signaling pathway to which the oligophrenin 1 protein belongs, wherein said drugs are tested on transgenic non-human mammals, or cells in culture, that overexpress oligophrenin 1 protein as defined in claim 4 or express a native oligophrenin 1 protein as defined in claim 4 that has been rendered non-functional.

23. Drug selected by the method of claim 22.

24. Pharmaceutical composition containing a drug of claim 23 in association with a pharmaceutically acceptable carrier.

25. Method of preventing and/or treating neurological disorders resulting from defects in the oligophrenin 1 gene or in the oligophrenin 1 protein or in a homologous gene or protein thereof, which comprises administering to a subject in need of a such treatment an amount of a pharmaceutical composition of claim 18 or 24 effective to prevent and/or alleviate said neurological disorders.