JP2001327294A - Human sodium channel SCN12A - Google Patents

Abstract

(57) [Problem] To provide a novel sodium channel SCN12A, its gene and cDNA. SOLUTION: Sodium channel SCN12A or SCN12A having the amino acid sequence of SEQ ID NO: 2 or 4
-S, a human gene encoding them, the SCN12A gene present on the short arm of human chromosome 3 (3p23-p21.3), and the cDNA of this gene;
Or a polynucleotide having a base sequence of 3.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

FIELD OF THE INVENTION This application relates to a novel alpha subunit SCN12A of the sodium channel of the human nervous system.
And its splicing variants SCN12A-s, human genes encoding these molecules, and the cDNA of this gene.

[0002]

BACKGROUND OF THE INVENTION Membrane voltage-gated sodium channels are:
It is a membrane protein molecule present in the cell membrane of excitable cells such as nerves and muscles. Excitable cells perform their function by generating action potentials. For example, in the case of a nerve cell, information, which is a function of the nerve cell, is transmitted by the action potential generated in the cell body propagating through the axon. Sodium channels are protein molecules that are responsible for the initiation and propagation of action potentials together with potassium channels.

[0003] The membrane voltage-gated sodium channel is composed of a plurality of subunits, the main function of which is provided by the α subunit. To date, the existence of multiple α subunit genes has been demonstrated in mammals, and at least SCN1A in humans.
(2q24), SCN2A (2q23), SCN3A (2q24-q3
1), SCN4A (17q23.1-q25.3), SCN5A (3p2
1), SCN6A (2q21-q23), SCN8A (12q13),
SCN9A (2q24), 9 of SCN10A (3p22-q24)
Genes have been identified (parentheses are in human chromosome loci). However, not all of these genes have been cloned in their complete form.Some of them have only been reported as partial sequences, or human homologous genes have been identified from mouse or rat homologous sequences. Some exist.

In recent years, diseases caused by mutations in these genes have been identified in humans or animals. The α subunit of the sodium channel is not only a physiologically important molecule, but the mutation may cause disease. It has been revealed. For example, mutations in SCN4A have been shown to cause familial hyperkalemia periodic limb paralysis, and mutations in SCN5A cause long QT syndrome type 1 to develop. Although no corresponding disease in humans is known, it is also known that mouse mutant motor endplate disease (med) is caused by mutation of Scn8a.

[0005]

As described above, elucidation of the molecular biological entity and function of the membrane voltage-gated sodium channel is not only for understanding the physiological mechanism of humans but also for the direct cause of various diseases. It is extremely important in identifying Furthermore, inferring from its physiological function, substances that act on sodium channels may be drugs (eg, anesthetics, analgesics, muscle relaxants, etc.) that regulate the function of cells in which they are expressed. It is also important as a target for the development of new therapeutic drugs.

[0006] The inventors of this application cloned a gene for the purpose of confirming all sodium channels expressed in the human nervous system, and as a result, EST
(Expressed Sequence Tags) database with SCN
AA446997 registered as a partial sequence of 8A is SC
It was found that neither the partial sequence of N8A nor other known sodium channel genes differed from the sequence.
Then, the full-length cDNA containing the sequence of AA446997 was successfully cloned, and the novel sodium channel encoded by this cDNA was named SCN12A.

[0007] This application describes a novel sodium channel SCN12A discovered by the inventors, its gene and c-type.
It is an object to provide DNA specifically. Another object of the present application is to provide a polypeptide obtained by recombinant DNA technology and an antibody against SCN12A.

[0008]

This application discloses a human sodium channel SCN12A having the amino acid sequence of SEQ ID NO: 2 and a human sodium channel having the amino acid sequence of SEQ ID NO: 4. Provide channel SCN12A-s.

[0009] This application also discloses a human gene encoding SCN12A and SCN12A-s, which is located on the short arm of human chromosome 3 (3p23-p21.3).
Provide the gene.

[0010] Further, the present application relates to the cD of the human gene.
A polynucleotide having the nucleotide sequence of SEQ ID NO: 1 and the polynucleotide having the nucleotide sequence of SEQ ID NO: 3 are provided.

[0011] Further, this application relates to the SCN12A.
Alternatively, an antibody against SCN12A-s is provided.

Hereinafter, embodiments of the invention will be described, and the invention of this application will be described in more detail.

[0013]

DETAILED DESCRIPTION OF THE INVENTION The sodium channels SCN12A and SCN12A-s of the present invention are different transcripts expressed from the same gene, respectively.
A is composed of 1791 amino acids shown in SEQ ID NO: 2,
SCN12A-s consists of 1444 amino acids shown in SEQ ID NO: 4 (hereinafter, these two may be simply referred to as "SCN12A").

The SCN12A of the present invention is a known method, that is, a method of isolating human various organs (eg, brain, spleen, small intestine, placenta, spinal cord, etc., as confirmed in the following Examples). A method of preparing a peptide by chemical synthesis based on the amino acid sequence provided by
Alternatively, it can be obtained by a method of producing a recombinant DNA technique using the polynucleotide provided by this application. For example, when obtaining a protein by recombinant DNA technology, RNA is prepared by in vitro transcription from a vector having the polynucleotide (SEQ ID NO: 1 or 3) of the present invention, and in vitro translation is performed using this as a template. The SCN12A protein can be obtained. In addition, if the translation region of the polynucleotide is recombined into an appropriate expression vector by a known method, and this recombinant vector is used to transform Escherichia coli, Bacillus subtilis, yeast, animal or plant cells, etc., these transformants can produce a large amount of protein. Can be expressed.

When the SCN12A of the present invention is produced by in vitro translation, the translation region of the polynucleotide of the present invention is recombined into a vector having an RNA polymerase promoter, a rabbit reticulocyte lysate containing an RNA polymerase corresponding to the promoter, or the like. It may be added to an in vitro translation system such as a wheat germ extract. Examples of the RNA polymerase promoter include T7, T3, SP6 and the like. Vectors containing these RNA polymerase promoters include pKA1, pCDM8, pT
3 / T7 18, pT7 / 3 19, pBluescript II
And the like.

When the SCN12A of the present invention is expressed in a microorganism such as Escherichia coli, an origin, a promoter, a ribosome binding site, and a DNA capable of replicating in the microorganism are used.
The translation region of the polynucleotide of the present invention may be recombined with an expression vector having a cloning site, a terminator, and the like to prepare an expression vector, a host cell may be transformed with the expression vector, and the transformant may be cultured. At this time, if a start codon and a stop codon are added before and after an arbitrary translation region, a protein fragment containing the arbitrary region can be obtained. Alternatively, it can be expressed as a fusion protein with another protein. By cleaving this fusion protein with an appropriate protease, SC
Only N12A can be obtained. Examples of expression vectors for Escherichia coli include pUC system, pBluescript II, pET expression system, pGEX expression system, and the like.

When the SCN12A of the present invention is to be expressed in eukaryotic cells, the translation region of the polynucleotide of the present invention is assembled into an expression vector for eukaryotic cells having a promoter, a splicing region, a poly (A) addition site and the like. Introduce into eukaryotic cells. As an expression vector, pKA
1, pCDM8, pSVK3, pMSG, pSVL, p
BK-CMV, pBK-RSV, EBV vector, pR
S and pYES2 can be exemplified. As eukaryotic cells,
Mammalian cultured cells such as monkey kidney cells COS7 and Chinese hamster ovary cells CHO, budding yeast, fission yeast, silkworm cells, Xenopus egg cells and the like are generally used, but are not limited thereto. In order to introduce the expression vector into eukaryotic cells, known methods such as an electroporation method, a calcium phosphate method, a liposome method, and a DEAE dextran method can be used.

After expressing a protein in a prokaryotic cell or a eukaryotic cell by the above-mentioned method, in order to isolate and purify the target protein from the culture, a known separation operation is combined. For example, treatment with a denaturant such as urea or a surfactant, ultrasonic treatment, enzymatic digestion, salting out or solvent precipitation, dialysis, centrifugation, ultrafiltration, gel filtration, SDS-PAG
E, isoelectric focusing, ion exchange chromatography,
Hydrophobic chromatography, affinity chromatography, reverse phase chromatography and the like.

The SCN12A of the present invention includes a peptide fragment (5 amino acid residues or more) containing any partial sequence in the amino acid sequence of SEQ ID NO: 2 or 4. These peptide fragments can be used as antigens for producing antibodies. The SCN12A of the present invention includes:
Fusion proteins with any other proteins are also included.

The gene of the present invention is a human gene encoding the above-mentioned SCN12A, which is present on the third short arm of human chromosome (3p23-p21.3). This gene can be isolated from an existing genomic library, for example, using the polynucleotide of the present invention or a partial sequence thereof as a probe.

The polynucleotide (cDNA) of the present invention
Can be cloned, for example, from cDNA derived from human cells. cDNA is poly A extracted from human cells
⁺ Synthesize using RNA as template. Okayama-Berg method (Mol. Cell. Biol. 2: 161-170, 1982), Guble
The r-Hoffman method (J. Gene 25: 263-269, 1983), the capping method (Gene 150: 243-250, 1994) and the like can be used. A commercially available human cDNA library can also be used. The cDNA library of the present invention
To clone NA, an oligonucleotide may be synthesized based on the nucleotide sequence of an arbitrary portion of the cDNA of the present invention, and screening may be performed by colony or plaque hybridization by a known method using the oligonucleotide as a probe. . Also, the target cDN
Oligonucleotides that hybridize to both ends of the A fragment can be synthesized, and using the oligonucleotides as primers, the cDNA of the present invention can be prepared from mRNA isolated from human cells by RT-PCR.

In general, polymorphisms due to individual differences are frequently observed in human genes. Therefore, the cDNA of SEQ ID NO: 1 or 3 in which one or more nucleotides are added, deleted and / or substituted by other nucleotides are also included in the polynucleotide of the present invention. Similarly, SCN12A in which one or more amino acids have been added, deleted and / or substituted by another amino acid resulting from these base changes also have the activity of SCN12A having the amino acid sequence of SEQ ID NO: 2 or 4. As long as it has, it is included in the present invention.

Furthermore, the polynucleotide of the present invention includes a DNA fragment (10 bp or more) containing any partial sequence of the nucleotide sequence of SEQ ID NO: 1 or 3. It also includes a DNA fragment consisting of a sense strand and an antisense strand. These DNA fragments can be used as probes for gene diagnosis.

The antibody of the present invention comprises SCN12A itself,
Alternatively, it can be obtained as a polyclonal antibody or a monoclonal antibody by a known method using a partial peptide thereof as an antigen.

Next, as an example, the SCN12A of the present invention will be described.
The following shows the details of the acquisition process and the results of its characteristics.

[0026]

EXAMPLES (1) Purification of human autopsy brain mRNA 2 g of autopsy brain slice from a patient with non-neurological disease was added to 20 mL of TRIzol reagent (Gibc
o-BRL) with a homogenizer and incubated at 30 ° C. for 15 minutes. Next, 4 mL of chloroform was added, stirred, and then incubated at 30 ° C. for 30 minutes. The mixture was centrifuged at 12,000 g for 15 minutes, the aqueous layer was separated, and total RNA was extracted by ethanol precipitation. T using Dynabeads mRNA purification kit (DYNAL)
The poly ⁺ RNA (mRNA) fraction was purified from otal RNA. (2) RT-PCR Using Superscript II reverse transcriptase (manufactured by Gibco-BRL), single-stranded DNA was synthesized using the mRNA purified in (1) as a template and random primers. This one
SC by nested PCR using single-stranded DNA as a template
1832 bp of the partial sequence of N12A was obtained. The primers used for this PCR were, as shown in FIG.
High homology between 446997 and known sodium channel α subunits (human H and rat R);
The design was based on the well conserved part of the sequence. Specifically, the following primers were used: Est-f1 (SEQ ID NO: 5)
Oligonucleotide), Est-f2 (oligonucleotide of SEQ ID NO: 6), Na-r1 (oligonucleotide of SEQ ID NO: 7), Na-r2 (oligonucleotide of SEQ ID NO: 8). (3) RACE Primers were synthesized based on the base sequence of the RT-PCR product obtained in the above (2), and 5′-RACE and 3′-RACE were obtained using Marathon-ready cDNA (Clontech) as a template.
And cDNA with 5 'and 3' adapters
I got The primer for 5'-RACE was Race-5.
(Oligonucleotide of SEQ ID NO: 9)
Race-3a (oligonucleotide of SEQ ID NO: 10) and Race-3b (oligonucleotide of SEQ ID NO: 11) were used as primers for ACE. The adapter primer is AP
1 (oligonucleotide of SEQ ID NO: 12) and AP2 (oligonucleotide of SEQ ID NO: 13) were used. (4) Subcloning The cDNA fragment of SCN12A obtained by RT-PCR and RACE was transformed into a plasmid vector pBleuscript IISK.
Subcloned into (+). (5) Sequencing The plasmid of the above (4) was propagated and purified, subjected to a sequencing reaction by a dye terminator and a dye primer method, and the reaction product was subjected to a fluorescent sequencer 377A (Pe
rkin Elmer / ABI). (6) Northern blot analysis Using an RNA blot membrane (Clontech) prepared from human various organ poly A ⁺ RNA, SCN12 was used.
Northern blot was performed using the AcDNA fragment as a probe. The probe was labeled with [α- ³² P] dCTP.
Hybridization results were analyzed with a Storm 830 image analyzer (Molecukar Dynamics). (7) In situ hybridization analysis In situ hybridization (in situ hybridization) using a cDNA clone as a probe and rat organs
u Hybridization), the detailed expression site was examined. (8) Identification of chromosomal locus Using the cDNA clone as a probe, the chromosomal locus was identified by fluorescence in situ hybridization (FISH). (9) Production of full-length cDNA expression clone SCN12A containing full-length ORF by RT-PCR using human spleen poly A ⁺ RNA (Clontech) as a template
And SCN12A-s, and these were used as plasmid vectors pSP64TR (pSP64Poly (A) manufactured by Promega).
Was modified (Fig. 2). The SCN12A PCR primer was SEQ ID NO: 14
And 15 oligonucleotides, also SCN12A-s
Oligonucleotides of SEQ ID NOS: 14 and 16 were used as PCR primers. (10) Results and discussion RACE results, two different types of cD
An NA fragment was obtained. The difference on the 3 'side is alternative splicing
it is conceivable that. Of the two cDNAs reconstructed from these fragments, the longer one was SCN12A cDNA and the shorter one was SCN12A-scDNA (FIG. 3). SC
N12A cDNA is 6528 bp in length, has an ORF of 5373 bp, and encodes a protein of 1791 amino acid residues.
The SCN12A-scDNA is 5728 bp in total length and 4332 bp OR.
It has an F and encodes a protein of 1444 amino acid residues. Both proteins have sodium channel α
Four transmembrane domains common to the subunits are identified.

Table 1 shows the known sodium channel α
The homology with the subunit, the chromosomal locus, the expression site and the relationship with the disease are also shown. The homology with the known α-subunit is about 37-73%, and since it has a considerably distant relationship from those previously known, SCN12A
Has a different function from the known α subunit.

SCN12A is a known tetrodotoxin resis
Scn, a tant (TTX-R) sodium channel
Similar to 10a / PN3 / SNS and NaN / SNS2, the presence of a serine amino acid residue in the SS2 segment of the transmembrane domain suggested that it was a TTX-R sodium channel.

The chromosomal locus of the SCN12A gene was mapped to the short arm of human chromosome 3 (3p23-p21.3) (FIG. 4). This position corresponds to the known α subunit SCN5A (3p
21) and sodium channel α in 3p21-p23
This suggests that a cluster of subunit genes may exist.

[0030]

[Table 1]

The results of Northern blot analysis are as shown in FIG. 5, and the SCN12A mRNA size was about 7.0.
With Kb, expression in the brain, spleen, small intestine, placenta and spinal cord was observed. Expression in other tissues was below the detection sensitivity in Northern blots. As also shown in Table 1, known sites of expression of the sodium channel α subunit are nerve tissues such as brain, spinal cord and dorsal root ganglion, and muscle tissues such as skeletal muscle, cardiac muscle and uterus. On the other hand, SCN12A
Expression was observed in the brain, spinal cord, spleen, small intestine, and placenta, but almost no expression was observed in skeletal muscle, extensor muscle, and uterus, confirming that the expression pattern was different from that of known genes.

In order to examine the expression site in more detail, in situ hybridization was performed using a rat. As a result, SCN12A was found to contain olfactory bulb, granular layer of cerebellum, dentate gyrus of hippocampus, ependymal cells of spinal cord, embryo of spleen Strong expression was shown in the vegetative germ layers of the center and placenta (FIG. 6). In addition, as a result of RT-PCR using a human cell line,
CN12A is expressed in both neurons and glial cells in the nervous system, and no RT-PCR band was observed in the dorsal root ganglion (DRG) due to human material problems. SCN12A was also expressed in the dorsal root ganglion (FIG. 7).

[0033]

As described in detail above, the present application provides a novel sodium channel α subunit and its cDNA. These inventions will greatly contribute to elucidation of the physiological mechanism involving excitable cells, identification of the causes of various human diseases, and development of new therapeutic agents.

[0034]

[Sequence List] SEQUENCE LISTING <110> Japan Science and Technology Corporation <120> Human Sodium Channel SCN12A <130> NP00167 <140> <141> <160> 16 <170> PatentIn Ver. 2.1 <210> 1 <211> 6528 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (200) .. (5575) <300> <301> Soen-Yong Jeong et al. <302> Identification of a Novel Human Voltage-Gated Sodium Channel α Subunit Gene, SCN12A <303> Biochem. Biophys. Res. Commun. <304> 267 <305> 1 <306> 262-270 <307> 2000 <308> GenBank AF109737 <309> 2000-01-15 <400> 1 tcctgctata cccacagtgg tggtcatctc ttctgatctt cacagccaat cagctcccaa 60 ggcccctgac ctcagctcag cttttgtaga tccttatgac accatccttt aagactggaa 120 tcctagggca ggctgtttta ttcccgcctc ctgaggcctt tctgaggatc tgtggcttgt 180 ctctgtcctg agggtgaag atg gat gac aga tgc tac cca gta atc ttt cca 232 Met Asp Asp Arg Cys Tyr Pro Val Ile Phe Pro 1 5 10 gat gag cgg aat ttc cgc ccc ttc act tcc gac tct ctg gct gca att 280 Asp Glu Arg Asn Phe Arg Pro Phe Thr Ser Asp Ser Leu Ala Ala Ile 15 20 25 gag aag cgg att gcc atc caa aag gag aaa aag aag tct aaa gac cag 328 Glu Lys Arg Ile Ala Ile Gln Lys Glu Lys Lys Lys Ser Lys Asp Gln 30 35 40 aca gga gaa gta ccc cag cct cgg cct cag ctt gac cta aag gcc tcc 376 Thr Gly Glu Val Pro Gln Pro Arg Pro Gln Leu Asp Leu Lys Ala Ser 45 50 55 agg aag ttg ccc aag ctc tat ggc gac att cct cgt gag ctc ata gga 424 Arg Lys Leu Pro Lys Leu Tyr Gly Asp Ile Pro Arg Glu Leu Ile Gly 60 65 70 75 aag cct ctg gaa gac ttg gac cca ttc tac cga aat cat aag aca ttt 472 Lys Pro Leu Glu Asp Leu Asp Pro Phe Tyr Arg Asn His Lys Thr Phe 80 85 90 atg gtg tta aac aga aag agg aca atc tac cgc ttc agt gcc aag cat 520 Met Val Leu Asn Arg Lys Arg Thr Ile Tyr Arg Phe Ser Ala Lys His 95 100 105 gcc ttg ttc att ttt ggg cct ttc aat tca atc aga agt tta gcc att 568 Ala Leu Phe Ile Phe Gly Pro Phe Asn Ser Ile Arg Ser Leu Ala Ile 110 115 120 aga gtc tca gtc cat tca ttg ttc agc atg ttc att atc gg 616 Arg Val Ser Val His Ser Leu Phe Ser Met Phe Ile Ile Gly Thr Val 125 130 135 atc atc aac tgc gtg ttc atg gct aca ggg cct gct aaa aac agc aac 664 Ile Ile Asn Cys Val Phe Met Ala Thr Gly Pro Ala Lys Asn Ser Asn 140 145 150 155 agt aac aat act gac att gca gag tgt gtc ttc act ggg att tat att 712 Ser Asn Asn Thr Asp Ile Ala Glu Cys Val Phe Thr Gly Ile Tyr Ile 160 165 170 ttt gaa gct ttg att aaa ata ttg gca aga ggt ttc att ctg gat gag 760 Phe Glu Ala Leu Ile Lys Ile Leu Ala Arg Gly Phe Ile Leu Asp Glu 175 180 185 ttt tct ttc ctt cga gat cca tgg aac tgg ctg gac tcc att gtc att 808 Arg Asp Pro Trp Asn Trp Leu Asp Ser Ile Val Ile 190 195 200 gga ata gcg att gtg tca tat att cca gga atc acc atc aaa cta ttg 856 Gly Ile Ala Ile Val Ser Tyr Ile Pro Gly Ile Thr Ile Lys Leu Leu 205 210 215 ccc ctg cgt acc ttc cgt gtg ttc aga gct ttg aaa gca att tca gta 904 Pro Leu Arg Thr Phe Arg Val Phe Arg Ala Leu Lys Ala Ile Ser Val 220 225 230 235 gtt tca cgt ctg aag gtc atc gtg ggg gcc ttg tct gtg aag 952 Val Ser Arg Leu Lys Val Ile Val Gly Ala Leu Leu Arg Ser Val Lys 240 245 250 aag ctg gtc aac gtg att atc ctc acc ttc ttt tgc ctc agc atc ttt 1000 Lys Leu Val Asn Val Ile Ile Leu Thr Phe Phe Cys Leu Ser Ile Phe 255 260 265 gcc ctg gta ggt cag cag ctc ttc atg gga agt ctg aac ctg aaa tgc 1048 Ala Leu Val Gly Gln Gln Leu Phe Met Gly Ser Leu Asn Leu Lys Cys 270 275 275 atc tcg ag aaa aat atc agt aac ccg gaa gct tat gac cat 1096 Ile Ser Arg Asp Cys Lys Asn Ile Ser Asn Pro Glu Ala Tyr Asp His 285 290 295 tgc ttt gaa aag aaa gaa aat tca cct gaa ttc aaa atg tgt ggc atc 1 Phe Glu Lys Lys Glu Asn Ser Pro Glu Phe Lys Met Cys Gly Ile 300 305 310 315 tgg atg ggt aac agt gcc tgt tcc ata caa tat gaa tgt aag cac acc 1192 Trp Met Gly Asn Ser Ala Cys Ser Ile Gln Tyr Glu Cys Lys His Thr 320 325 330 aaa att aat cct gac tat aat tat acg aat ttt gac aac ttt ggc tgg 1240 Lys Ile Asn Pro Asp Tyr Asn Tyr Thr Asn Phe Asp Asn Phe Gly Trp 335 340 345 345 tct ttt ctt gcc atg ttc cgg ctg atg acc caa gat tcc tgg gag aag 1288 Ser Phe Leu Ala Met Phe Arg Leu Met Thr Gln Asp Ser Trp Glu Lys 350 355 360 ctt tat caa cag acc ctg cgt act act ggg ctc tac tca gtc ttc ttc 1336 Leu Tyr Gln Gln Thr Leu Arg Thr Thr Gly Leu Tyr Ser Val Phe Phe 365 370 375 ttc att gtg gtc att ttc ctg ggc tcc ttc tac ctg att aac tta acc 1384 Phe Ile Val Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Thr 380 385 390 ctg gct gtt gtt acc atg gca tat gag gag cag aac aag aat gta gct 1432 Leu Ala Val Val Thr Met Ala Tyr Glu Glu Gln Asn Lys Asn Val Ala 400 405 410 gca gag ata gag gcc aag gaa aag atg ttt cag gaa g cc cag cag ctg 1480 Ala Glu Ile Glu Ala Lys Glu Lys Met Phe Gln Glu Ala Gln Gln Leu 415 420 425 tta aag gag gaa aag gag gct ctg gtt gcc atg gga att gac aga agt 1528 Leu Lys Glu Glu Lys Glu Alu Ala Met Gly Ile Asp Arg Ser 430 435 440 tca ctt act tcc ctt gaa aca tca tat ttt acc cca aaa aag aga aag 1576 Ser Leu Thr Ser Leu Glu Thr Ser Tyr Phe Thr Pro Lys Lys Arg Lys 445 450 455 ctc ttt ggt aat aag aaa agg aag tcc ttc ttt ttg aga gag tct ggg 1624 Leu Phe Gly Asn Lys Lys Arg Lys Ser Phe Phe Leu Arg Glu Ser Gly 460 465 470 470 475 aaa gac cag cct cct ggg tca gat tct gat gaa gat tgc caaaaa Lys Asp Gln Pro Pro Gly Ser Asp Ser Asp Glu Asp Cys Gln Lys Lys 480 485 490 cca cag ctc cta gag caa acc aaa cga ctg tcc cag aat cta tca ctg 1720 Pro Gln Leu Leu Glu Gln Thr Lys Arg Leu Ser Gln Asn Leu Ser Leu 495 500 505 gac cac ttt gat gag cat gga gat cct ctc caa agg cag aga gca ctg 1768 Asp His Phe Asp Glu His Gly Asp Pro Leu Gln Arg Gln Arg Ala Leu 510 515 520 agt gcc gtc agc atc ctc acc at c acc atg aag gaa caa gaa aaa tca 1816 Ser Ala Val Ser Ile Leu Thr Ile Thr Met Lys Glu Gln Glu Lys Ser 525 530 535 caa gag cct tgt ctc cct tgc gga gaa aac ctg gca tcc aag tac ctc 1864 Gln Glu Pro Cys Leu Pro Cys Gly Glu Asn Leu Ala Ser Lys Tyr Leu 540 545 550 555 gtg tgg aac tgt tgc ccc cag tgg ctg tgc gtt aag aag gtc ctg aga 1912 Val Trp Asn Cys Cys Pro Gln Trp Leu Cys Val Lys Lys Val Leg Arg 565 570 act gtg atg act gac ccg ttt act gag ctg gcc atc acc atc tgc atc 1960 Thr Val Met Thr Asp Pro Phe Thr Glu Leu Ala Ile Thr Ile Cys Ile 575 580 585 atc atc aac act gtc ttc ttg gcc atg gag cat cac aag atg gag gcc 2008 Ile Ile Asn Thr Val Phe Leu Ala Met Glu His His Lys Met Glu Ala 590 595 600 agt ttt gag aag atg ttg aat ata ggg aat ttg gtt ttc act agc att 2056 Ser Phe Glu Lys Met Leu Asn Ile Gly Asn Leu Val Phe Thr Ser Ile 605 610 615 ttt ata gca gaa atg tgc cta aaa atc att gcg ctc gat ccc tac cac 2104 Phe Ile Ala Glu Met Cys Leu Lys Ile Ile Ala Leu Asp Pro Tyr His 620 625 630 630 tactt t cgc cga ggc tgg aac att ttt gac agc att gtt gct ctt ctg 2152 Tyr Phe Arg Arg Gly Trp Asn Ile Phe Asp Ser Ile Val Ala Leu Leu 640 645 650 agt ttt gca gat gta atg aac tgt gta ctt caa aag aga cca 2200 Ser Phe Ala Asp Val Met Asn Cys Val Leu Gln Lys Arg Ser Trp Pro 655 660 665 ttc ttg cgt tcc ttc agg gtg ctc agg gtc ttc aag tta gcc aaa tcc 2248 Phe Leu Arg Ser Phe Arg Val Leu Arg Val Phe Leu Ala Lys Ser 670 675 680 tgg cca act ttg aac aca cta att aag ata atc ggc aac tct gtc gga 2296 Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile Gly Asn Ser Val Gly 685 690 695 gcc ctt gga aac ctg act gtg gtc ctg gtc att gtg atc ttt att ttc 2344 Ala Leu Gly Asn Leu Thr Val Val Leu Val Ile Val Ile Phe Ile Phe 700 705 710 715 tca gta gtt ggc atg cag ctt ttt ggc cgt agc ttc aat tcc caa aag 2392 Gly Met Gln Leu Phe Gly Arg Ser Phe Asn Ser Gln Lys 720 725 730 agt cca aaa ctc tgt aac ccg aca ggc ccg aca gtc tca tgt tta cgg 2440 Ser Pro Lys Leu Cys Asn Pro Thr Gly Pro Thr Val Ser Cys Leu Arg 735 740 745 cac tgg cac atg ggg gat ttc tgg cac tcc ttc cta gtg gta ttc cgc 2488 His Trp His Met Gly Asp Phe Trp His Ser Phe Leu Val Val Phe Arg 750 755 760 atc ctc tgc ggg gaa tgg atg gaa gaa tgt atg caa gaa 2536 Ile Leu Cys Gly Glu Trp Ile Glu Asn Met Trp Glu Cys Met Gln Glu 765 770 775 gcg aat gca tca tca tca ttg tgt gtt att gtc ttc ata ttg atc acg 2584 Ala Sern As Ser Ser Val Ile Val Phe Ile Leu Ile Thr 780 785 790 795 gtg ata gga aaa ctt gtg gtg ctc aac ctc ttc att gcc tta ctg ctc 2632 Val Ile Gly Lys Leu Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu 800 t 805 810 810 ttt agc aat gag gaa aga aat gga aac tta gaa gga gag gcc 2680 Asn Ser Phe Ser Asn Glu Glu Arg Asn Gly Asn Leu Glu Gly Glu Ala 815 820 825 agg aaa act aaa gtc cag tta gca ctg gat cga ttccg 2728 Arg Lys Thr Lys Val Gln Leu Ala Leu Asp Arg Phe Arg Arg Ala Phe 830 835 840 tgt ttt gtg aga cac act ctt gag cat ttc tgt cac aag tgg tgc agg 2776 Cys Phe Val Arg His Thr Leu Glu His Phe Cys His Lys Trp Cys Arg 845 850 855 aag caa aac tta cca cag caa aaa gag gtg gca gga ggc tgt gct gca 2824 Lys Gln Asn Leu Pro Gln Gln Lys Glu Val Ala Gly Gly Cys Ala Ala 860 865 870 870 875 caa agc aaa gac att ccc ctg gtc atg gag atg aaa agg ggc tca 2872 Gln Ser Lys Asp Ile Ile Pro Leu Val Met Glu Met Lys Arg Gly Ser 880 885 890 gag acc cag gag gag ctt ggt ata cta acc tct gta cca aag acc ctg 2920 Glu Thr Gln Glu Glu Leu Gly Ile Leu Thr Ser Val Pro Lys Thr Leu 895 900 905 ggc gtc agg cat gat tgg act tgg ttg gca cca ctt gcg gag gag gaa 2968 Gly Val Arg His Asp Trp Thr Trp Leu Ala Pro Leu Ala Glu Glu Glu 910 915 920 gat gac gtt gaa ttt tct ggt gaa gat aat gca cag cgc atc aca caa 3016 Asp Asp Val Glu Phe Ser Gly Glu Asp Asn Ala Gln Arg Ile Thr Gln 925 930 935 cct gag cct gaa caa cag gcc tat gag cag gag aac aag aag 3064 Pro Glu Pro Glu Gln Gln Ala Tyr Glu Leu His Gln Glu Asn Lys Lys 940 945 950 955 ccc acg agc cag aga gtt caa agt gtg gaa att gac atg ttc tct gaa 3112 Pro Thr Ser Gln Arg Val Gln Ser Val Glu Ile Asp Met Phe Ser Glu 960 965 970 gat gag cct cat ctg acc ata cag gat ccc cga aag aag tct gat gtt 3160 Asp Glu Pro His Leu Thrrl Ile Gln Asp Pro Arg Lys Lys Ser Asp Val 975 980 985 acc agt ata cta tca gaa tgt agc acc att gat ctt cag gat ggc ttt 3208 Thr Ser Ile Leu Ser Glu Cys Ser Thr Ile Asp Leu Gln Asp Gly Phe 990 995 1000 gga tgg tta cct gag atg gtt ccc aaa aag caa cca gag aga tgt ttg 3256 Gly Trp Leu Pro Glu Met Val Pro Lys Lys Gln Pro Glu Arg Cys Leu 1005 1010 1015 ccc aaa ggc ttt ggt tgc tgc ttt cca tgc tgt agc gtg gac aag aga 3304 Pro Lys Gly Phe Gly Cys Cys Phe Pro Cy Cys Cy Ser Val Asp Lys Arg 1020 1025 1030 1035 aag cct ccc tgg gtc att tgg tgg aac ctg cgg aaa acc tgc tac caa 3352 Lys Pro Pro Trp Val Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Gln 1040 1045 1050 ata gtc aaacacg tgg ttt gag agc ttt att atc ttt gtg att ctg 3400 Ile Val Lys His Ser Trp Phe Glu Ser Phe Ile Ile Phe Val Ile Leu 1055 1060 1065 ctg agc agt ggg gca ctg ata ttt gaa gat gtt cac ctt gag aac caa 3448 Leu Ser Ser Gly Ala Leu Ile Phe Glu Asp Val His Leu Glu Asn Gln 1070 1075 1080 ccc aaa atc caa gaa tta cta aat tgt act gac att att ttt aca cat 3496 Pro Lys Ile Gln Glu Leu Leu Asn Cys Thr Asp Ile Ile Phe Thr His 1085 1090 1095 att ttt atc ctg gag atg gta cta aaa tgg gta gcc ttc gga ttt gga 3544 Ile Phe Ile Leu Glu Met Val Leu Lys Trp Val Ala Phe Gly Phe Gly 1100 1105 1110 1115 aag tat ttc acc gcc tgg tgc tgc ctt gat ttc atc att gtg att 3592 Lys Tyr Phe Thr Ser Ala Trp Cys Cys Leu Asp Phe Ile Ile Val Ile 1120 1125 1130 gtc tct gtg acc acc ctc att aac tta atg gaa ttg agg tcc ttc Val Thr Thr Leu Ile Asn Leu Met Glu Leu Lys Ser Phe Arg 1135 1140 1145 act cta cga gca ctg agg cct ctt cgt gcg ctg tcc cag ttt gaa gga 3688 Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Gln Phe Glu Gly 1150 1155 1160 atg aag gtg gtg gtc aat gct ctc ata ggt gcc ata cct gcc att ctg 3736 Met Lys Val Val Val Asn Ala Leu Ile Gly Ala Ile Pro Ala Ile Leu 1165 1170 1175 aat gtt ttg ct t gtc tgc ctc att ttc tgg ctc gta ttt tgt att ctg 3784 Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu Val Phe Cys Ile Leu 1180 1185 1190 1195 gga gta tac ttc ttt tct gga aaa ttt ggg aa tgc atg 3832 Gly Val Tyr Phe Phe Ser Gly Lys Phe Gly Lys Cys Ile Asn Gly Thr 1200 1205 1210 gac tca gtt ata aat tat acc atc att aca aat aaa agt caa tgt gaa 3880 Asp Ser Val Ile Asn Tyr Thr Ile Ile Thr Asn Lys Ser Gln Cys Glu 1215 1220 1225 agt ggc aat ttc tct tgg atc aac cag aaa gtc aac ttt gac aat gtg 3928 Ser Gly Asn Phe Ser Trp Ile Asn Gln Lys Val Asn Phe Asp Asn Val 1230 1235 1240 gga aat gct tac ctcgct caa gtg gca aca ttt aag ggc tgg 3976 Gly Asn Ala Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp 1245 1250 1255 atg gat att ata tat gca gct gtt gat tcc aca gag aaa gaa caa cag 4024 Met Asyr Ile Ala Ala Val Asp Ser Thr Glu Lys Glu Gln Gln 1260 1265 1270 1275 cca gag ttt gag agc aat tca ctc ggt tac att tac ttc gta gtc ttt 4072 Pro Glu Phe Glu Ser Asn Ser Leu Gly Tyr Ile Tyr Phe Val Val Phe 1280 1285 1290 atc atc ttt ggc tca ttc ttc act ctg aat ctc ttc att ggc gtt atc 4120 Ile Ile Ple Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile 1295 1300 1305 att gac aac ttcac cag aaa aag tta ggt ggc caa gac att 4168 Ile Asp Asn Phe Asn Gln Gln Gln Lys Lys Leu Gly Gly Gln Asp Ile 1310 1315 1320 ttt atg aca gaa gaa cag aag aaa tac tat aat gca atg aaa aaa 4tta Phe Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu 1325 1330 1335 gga tcc aaa aaa cct caa aaa ccc att cca cgg cct ctg aac aaa tgt 4264 Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Cys 1340 1345 1350 1355 caa ggt ctc gtg ttc gac ata gtc aca agc cag atc ttt gac atc atc 4312 Gln Gly Leu Val Phe Asp Ile Val Thr Ser Gln Ile Phe Asp Ile Ile 1360 1365 1370 atc ata agt ctc att atc cta ag atg atg atg gct gaa tca 4360 Ile Ile Ser Leu Ile Ile Leu Asn Met Ile Ser Met Met Ala Glu Ser 1375 1380 1385 tac aac caa ccc aaa gcc atg aaa tcc atc ctt gac cat ctc aac tgg 4408 Tyr A sn Gln Pro Lys Ala Met Lys Ser Ile Leu Asp His Leu Asn Trp 1390 1395 1400 gtc ttt gtg gtc atc ttt acg tta gaa tgt ctc atc aaa atc ttt gct 4456 Val Phe Val Val Ile Phe Thr Leu Glu Cys Leu Ile Lys Ile Phe Ala 1405 1410 1415 ttg agg caa tac tac ttc acc aat ggc tgg aat tta ttt gac tgt gtg 4504 Leu Arg Gln Tyr Tyr Phe Thr Asn Gly Trp Asn Leu Phe Asp Cys Val 1420 1425 1430 1435 gtc gtg ctt ctt tcct gtt atg att tct acc ttg gaa aat 4552 Val Val Leu Leu Ser Ile Val Ser Thr Met Ile Ser Thr Leu Glu Asn 1440 1445 1450 cag gag cac att cct ttc cct ccg acg ctc ttc aga att gtc cgc ttg 4600 Gln Glu His Ile Pro Phe Pro Pro Thr Leu Phe Arg Ile Val Arg Leu 1455 1460 1465 gct cgg att ggc cga atc ctg agg ctt gtc cgg gct gca cga gga atc 4648 Ala Arg Ile Gly Arg Ile Leu Arg Leu Val Arg Ala Ala Arg Gly Ile 1470 1475 agg act ctc ctc ttt gct ctg atg atg tcg ctt cct tct ctg ttc aac 4696 Arg Thr Leu Leu Phe Ala Leu Met Met Ser Leu Pro Ser Leu Phe Asn 1485 1490 1495 att ggt ctt cta ctc ttt ctg att a tg ttt atc tat gcc att ctg ggt 4744 Ile Gly Leu Leu Leu Phe Leu Ile Met Phe Ile Tyr Ala Ile Leu Gly 1500 1505 1510 1515 atg aac tgg ttt tcc aaa gtg aat cca gag tct gga atc gat gac ata 4792 Pet Asn Ser Lys Val Asn Pro Glu Ser Gly Ile Asp Asp Ile 1520 1525 1530 ttc aac ttc aag act ttt gcc agc agc atg ctc tgt ctc ttc cag ata 4840 Phe Asn Phe Lys Thr Phe Ala Ser Ser Met Leu Cys Leu Phe Gln 1535 1545 agc aca tca gca ggt tgg gat tcc ctg ctc agc ccc atg ctg cga tca 4888 Ser Thr Ser Ala Gly Trp Asp Ser Leu Leu Ser Pro Met Leu Arg Ser 1550 1555 1560 aaa gaa tca tgt aac tct tcc tca gaa acccc cct ggc ata 4936 Lys Glu Ser Cys Asn Ser Ser Ser Glu Asn Cys His Leu Pro Gly Ile 1565 1570 1575 gcc aca tcc tac ttt gtc agt tac att atc atc tcc ttt ctc att gtt 4984 Ala Thr Ser Tyr Phe Val Ser Tyr Ile Ile Ile Ser Phe Leu Ile Val 1580 1585 1590 1595 gtc aac atg tac att gct gtg att tta gag aac ttc aat aca gcc act 5032 Val Asn Met Tyr Ile Ala Val Ile Leu Glu Asn Phe Asn Thr Ala Thr 1 600 1605 1610 gaa gaa agt gag gac cct ttg ggt gaa gat gac ttt gac ata ttt tat 5080 Glu Glu Ser Glu Asp Pro Leu Gly Glu Asp Asp Phe Asp Ile Phe Tyr 1615 1620 1625 gaa gtg tgg gaa aga ttt gacca caa ttt atc aaa tat 5128 Glu Val Trp Glu Lys Phe Asp Pro Glu Ala Thr Gln Phe Ile Lys Tyr 1630 1635 1640 tct gcc ctt tct gac ttt gct gat gcc ttg cct gag cct ttg cgt gtc 5176 Ser Ala Leu Ser Asp Phe Ala Asp Ala Leu Pro Glu Pro Leu Arg Val 1645 1650 1655 gca aag cca aat aaa tat caa ttt cta gta atg gac ttg ccc atg gtg 5224 Ala Lys Pro Asn Lys Tyr Gln Phe Leu Val Met Asp Leu Pro Met Val 1660 1665 1670 1675 agt gaa gat cgc ctc cac tgc atg gat att ctt ttc gcc ttc acc gct 5272 Ser Glu Asp Arg Leu His Cys Met Asp Ile Leu Phe Ala Phe Thr Ala 1680 1685 1690 agg gta ctc ggt ggc tct gat ggc cta gat agt atg aaa 5320 Arg Val Leu Gly Gly Ser Asp Gly Leu Asp Ser Met Lys Ala Met Met 1695 1700 1705 gaa gag aag ttc atg gaa gcc aat cct ctc aag aag ttg tat gaa ccc 5368 Glu Glu Lys Phe Met Glu Ala Asn Pro Leu Lys Lys Leu Tyr Glu Pro 1710 1715 1720 ata gtc acc acc acc aag aga aag gaa gag gaa aga ggt gct gct att 5416 Ile Val Thr Thr Thr Thr Lys Arg Lys Glu Glu Glu Arg Gly Ala Ala Ile 1725 1730 1735 att caa aag gcc ttt cga aag tac atg atg aag gtg acc aag ggt gac 5464 Ile Gln Lys Ala Phe Arg Lys Tyr Met Met Lys Val Thr Lys Gly Asp 1740 1745 1750 1755 caa ggt gac caa aat gac ttg gaa aac ggg cct cat tca ctc cag 5512 Gln Gly Asp Gln Asn Asp Leu Glu Asn Gly Pro His Ser Pro Leu Gln 1760 1765 1770 act ctt tgc aat gga gac ttg tct agc ttt ggg gtg gcc aag ggc aag 5560 Thr Leu Cys Asn Gly Asp Leu Ser Ser Ghe Val Ala Lys Gly Lys 1775 1780 1785 gtc cac tgt gac tga gccctcacct ccacgcctac ctcatagctt cacagccttg 5615 Val His Cys Asp 1790 ccctcagcct ctgagctcca ggggtcagca gcttagtgta tcaacaggga gtggattcac 5675 caaattagcc attccatttt cttttctggc taaataaatg atatttcaat ttcattttaa 5735 atgatactta cagagatata agataaggct acttgacaac cagcggtact attttaataa 5795 ggaagaagac accaggaagg actgtaaaag gacattccaa t tttaggatt ggaatagttc 5855 aagccgggcg cagtggctca tgcctggaat cccagcactt tgagaggcca aggcaggtgg 5915 atcacgaggt caagagatcg agaccatcct ggccaaccat gatgaaactc cgtctctact 5975 aaaatacaaa aattagctgg gcatggttgc gtgcgcctgt agtcccagct acttgggagg 6035 ctgaggcagg agaatcgctt aaacctggga gacggaggtt gcagtgagcc aagatcgtgt 6095 cactgcactc cagcctggtg acagagtgag actctgtttc aaaaaagaaa agaaaagaaa 6155 catggttcaa attatatcta aacaaaaaag aataagaaac aaaaaacaca ttaaaatttt 6215 aagttgtatt ttctatgttt ctagatacat catttttgtt tgatattttc ctgatgcaag 6275 tatgtggttt atcacatgta gctcttttgc atgctaaatg aaaattcaag acttgccaat 6335 aaatgaatag cttattgcag acatttttta ccaacattaa ttattttggg tttgtttaaa 6395 acctagaggc acaatcttga cttgtcaatt actacccttt cacaagctac catctcagat 6455 atatatatat atataaattc aataaagctt tctgtttgtg ttccataaaa aaaaaaaaaa 6515 aaaaaaaaaa aaa 6528 <210> 2 <211> 1791 <212> PRT <213> Homo sapiens <400> 2 Met Asp Asp Arg Cys Tyr Pro Val Ile Phe Pro Asp Glu Arg Asn Phe 1 5 10 15 Arg Pro Phe Thr Ser Asp Ser Leu Ala Ala Ile Glu Lys Arg Ile Ala 20 25 30 Ile Gln Lys Glu Lys Lys Lys Ser Lys Asp Gln Thr Gly Glu Val Pro 35 40 45 Gln Pro Arg Pro Gln Leu Asp Leu Lys Ala Ser Arg Lys Leu Pro Lys 50 55 60 Leu Tyr Gly Asp Ile Pro Arg Glu Leu Ile Gly Lys Pro Leu Glu Asp 65 70 75 80 Leu Asp Pro Phe Tyr Arg Asn His Lys Thr Phe Met Val Leu Asn Arg 85 90 95 Lys Arg Thr Ile Tyr Arg Phe Ser Ala Lys His Ala Leu Phe Ile Phe 100 105 110 Gly Pro Phe Asn Ser Ile Arg Ser Leu Ala Ile Arg Val Ser Val His 115 120 125 Ser Leu Phe Ser Met Phe Ile Ile Gly Thr Val Ile Ile Asn Cys Val 130 135 140 Phe Met Ala Thr Gly Pro Ala Lys Asn Ser Asn Ser Asn Asn Thr Asp 145 150 155 160 Ile Ala Glu Cys Val Phe Thr Gly Ile Tyr Ile Phe Glu Ala Leu Ile 165 170 175 Lys Ile Leu Ala Arg Gly Phe Ile Leu Asp Glu Phe Ser Phe Leu Arg 180 185 190 Asp Pro Trp Asn Trp Leu Asp Ser Ile Val Ile Gly Ile Ala Ile Val 195 200 205 Ser Tyr Ile Pr o Gly Ile Thr Ile Lys Leu Leu Pro Leu Arg Thr Phe 210 215 220 Arg Val Phe Arg Ala Leu Lys Ala Ile Ser Val Val Ser Arg Leu Lys 225 230 235 240 Val Ile Val Gly Ala Leu Leu Arg Ser Val Lys Lys Leu Val Asn Val 245 250 255 Ile Ile Leu Thr Phe Phe Cys Leu Ser Ile Phe Ala Leu Val Gly Gln 260 265 270 Gln Leu Phe Met Gly Ser Leu Asn Leu Lys Cys Ile Ser Arg Asp Cys 275 280 285 285 Lys Asn Ile Ser Asn Pro Glu Ala Tyr Asp His Cys Phe Glu Lys Lys 290 295 300 Glu Asn Ser Pro Glu Phe Lys Met Cys Gly Ile Trp Met Gly Asn Ser 305 310 315 320 Ala Cys Ser Ile Gln Tyr Glu Cys Lys His Thr Lys Ile Asn Pro Asp 325 330 335 Tyr Asn Tyr Thr Asn Phe Asp Asn Phe Gly Trp Ser Phe Leu Ala Met 340 345 350 Phe Arg Leu Met Thr Gln Asp Ser Trp Glu Lys Leu Tyr Gln Gln Thr 355 360 365 Leu Arg Thr Thr Gly Leu Tyr Ser Val Phe Phe Phe Ile Val Val Ile 370 375 380 Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Thr Leu Ala Val Val Thr 385 390 395 400 400 Met Ala Tyr Glu Glu Gln Asn Lys Asn Val Ala Ala Glu Ile Glu Ala 405 410 415 Lys Glu Lys M et Phe Gln Glu Ala Gln Gln Leu Leu Lys Glu Glu Lys 420 425 430 Glu Ala Leu Val Ala Met Gly Ile Asp Arg Ser Ser Leu Thr Ser Leu 435 440 445 Glu Thr Ser Tyr Phe Thr Pro Lys Lys Arg Lys Leu Phe Gly Asn Lys 450 455 460 Lys Arg Lys Ser Phe Phe Leu Arg Glu Ser Gly Lys Asp Gln Pro Pro 465 470 475 480 Gly Ser Asp Ser Asp Glu Asp Cys Gln Lys Lys Pro Gln Leu Leu Glu 485 490 495 Gln Thr Lys Arg Leu Ser Gln Asn Leu Ser Leu Asp His Phe Asp Glu 500 505 510 His Gly Asp Pro Leu Gln Arg Gln Arg Ala Leu Ser Ala Val Ser Ile 515 520 525 Leu Thr Ile Thr Met Lys Glu Gln Glu Lys Ser Gln Glu Pro Cys Leu 530 535 540 Pro Cys Gly Glu Asn Leu Ala Ser Lys Tyr Leu Val Trp Asn Cys Cys 545 550 555 560 Pro Gln Trp Leu Cys Val Lys Lys Val Leu Arg Thr Val Met Thr Asp 565 570 575 Pro Phe Thr Glu Leu Ala Ile Thr Ile Cys Ile Ile Ile Asn Thr Val Val 580 585 590 Phe Leu Ala Met Glu His His Lys Met Glu Ala Ser Phe Glu Lys Met 595 600 605 Leu Asn Ile Gly Asn Leu Val Phe Thr Ser Ile Phe Ile Ala Glu Met 610 615 620 Cys Leu Lys Ile I le Ala Leu Asp Pro Tyr His Tyr Phe Arg Arg Gly 625 630 635 640 Trp Asn Ile Phe Asp Ser Ile Val Ala Leu Leu Ser Phe Ala Asp Val 645 650 655 Met Asn Cys Val Leu Gln Lys Arg Ser Trp Pro Phe Leu Arg Ser Phe 660 665 670 Arg Val Leu Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn 675 680 685 Thr Leu Ile Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Asn Leu 690 695 700 Thr Val Val Leu Val Ile Val Ile Phe Ile Phe Ser Val Val Gly Met 705 710 715 715 720 Gln Leu Phe Gly Arg Ser Phe Asn Ser Gln Lys Ser Pro Lys Leu Cys 725 730 735 Asn Pro Thr Gly Pro Thr Val Ser Cys Leu Arg His Trp His Met Gly 740 745 750 Asp Phe Trp His Ser Phe Leu Val Val Phe Arg Ile Leu Cys Gly Glu 755 760 765 Trp Ile Glu Asn Met Trp Glu Cys Met Gln Glu Ala Asn Ala Ser Ser 770 775 780 Ser Leu Cys Val Ile Val Phe Ile Leu Ile Thr Val Ile Gly Lys Leu 785 790 795 800 Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe Ser Asn 805 810 815 Glu Glu Arg Asn Gly Asn Leu Glu Gly Glu Ala Arg Lys Thr Lys Val 820 825 830 Gln Leu Ala LeuAsp Arg Phe Arg Arg Ala Phe Cys Phe Val Arg His 835 840 845 Thr Leu Glu His Phe Cys His Lys Trp Cys Arg Lys Gln Asn Leu Pro 850 855 860 Gln Gln Lys Glu Val Ala Gly Gly Cys Ala Ala Gln Ser Lys Asp Ile 865 870 875 880 Ile Pro Leu Val Met Glu Met Lys Arg Gly Ser Glu Thr Gln Glu Glu 885 890 895 Leu Gly Ile Leu Thr Ser Val Pro Lys Thr Leu Gly Val Arg His Asp 900 905 910 Trp Thr Trp Leu Ala Pro Leu Ala Glu Glu Glu Asp Asp Val Glu Phe 915 920 925 925 Ser Gly Glu Asp Asn Ala Gln Arg Ile Thr Gln Pro Glu Pro Glu Gln 930 935 940 Gln Ala Tyr Glu Leu His Gln Glu Asn Lys Lys Pro Thr Ser Gln Arg 945 950 955 960 Val Gln Ser Val Glu Ile Asp Met Phe Ser Glu Asp Glu Pro His Leu 965 970 975 Thr Ile Gln Asp Pro Arg Lys Lys Ser Asp Val Thr Ser Ile Leu Ser 980 985 990 Glu Cys Ser Thr Ile Asp Leu Gln Asp Gly Phe Gly Trp Leu Pro Glu 995 1000 1005 Met Val Pro Lys Lys Gln Pro Glu Arg Cys Leu Pro Lys Gly Phe Gly 1010 1015 1020 Cys Cys Phe Pro Cys Cys Ser Val Asp Lys Arg Lys Pro Pro Trp Val 1025 1030 1035 1040 Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Gln Ile Val Lys His Ser 1045 1050 1055 Trp Phe Glu Ser Phe Ile Ile Phe Val Ile Leu Leu Ser Ser Gly Ala 1060 1065 1070 Leu Ile Phe Glu Asp Val His Leu Glu Asn Gln Pro Lys Ile Gln Glu 1075 1080 1085 Leu Leu Asn Cys Thr Asp Ile Ile Phe Thr His Ile Phe Ile Leu Glu 1090 1095 1100 Met Val Leu Lys Trp Val Ala Phe Gly Phe Gly Lys Tyr Phe Thr Ser 1105 1110 1115 1120 Ala Trp Cys Cys Leu Asp Phe Ile Ile Val Ile Val Ser Val Thr Thr 1125 1130 1135 Leu Ile Asn Leu Met Glu Leu Lys Ser Phe Arg Thr Leu Arg Ala Leu 1140 1145 1150 Arg Pro Leu Arg Ala Leu Ser Gln Phe Glu Gly Met Lys Val Val Val 1155 1160 1165 Asn Ala Leu Ile Gly Ala Ile Pro Ala Ile Leu Asn Val Leu Leu Val 1170 1175 1180 Cys Leu Ile Phe Trp Leu Val Phe Cys Ile Leu Gly Val Tyr Phe Phe 1185 1190 1195 1200 Ser Gly Lys Phe Gly Lys Cys Ile Asn Gly Thr Asp Ser Val Ile Asn 1205 1210 1215 Tyr Thr Ile Ile Thr Asn Lys Ser Gln Cys Glu Ser Gly Asn Phe Ser 1220 1225 1230 Trp Ile Asn Gln Lys Val Asn Phe Asp Asn Val Gly Asn Ala Tyr Leu 1235 1240 1245 Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Ile Tyr 1250 1255 1260 Ala Ala Val Asp Ser Thr Glu Lys Glu Gln Gln Pro Glu Phe Glu Ser 1265 1270 1275 1280 Asn Ser Leu Gly Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser 1285 1290 1295 Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn 1300 1305 1310 Gln Gln Gln Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu Glu 1315 1320 1325 Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro 1330 1335 1340 Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Cys Gln Gly Leu Val Phe 1345 1350 1355 1360 Asp Ile Val Thr Ser Gln Ile Phe Asp Ile Ile Ile Ile Ser Leu Ile 1365 1370 1375 Ile Leu Asn Met Ile Ser Met Met Ala Glu Ser Tyr Asn Gln Pro Lys 1380 1385 1390 Ala Met Lys Ser Ile Leu Asp His Leu Asn Trp Val Phe Val Val Ile 1395 1400 1405 Phe Thr Leu Glu Cys Leu Ile Lys Ile Phe Ala Leu Arg Gln Tyr Tyr 1410 1415 1420 Phe Thr Asn Gly Trp Asn Leu Phe Asp Cys Val Val Val Leu Leu Ser 1425 1430 1435 1440 Ile Va l Ser Thr Met Ile Ser Thr Leu Glu Asn Gln Glu His Ile Pro 1445 1450 1455 Phe Pro Pro Thr Leu Phe Arg Ile Val Arg Leu Ala Arg Ile Gly Arg 1460 1465 1470 Ile Leu Arg Leu Val Arg Ala Ala Arg Gly Ile Arg Thr Leu Leu Phe 1475 1480 1485 Ala Leu Met Met Ser Leu Pro Ser Leu Phe Asn Ile Gly Leu Leu Leu 1490 1495 1500 Phe Leu Ile Met Phe Ile Tyr Ala Ile Leu Gly Met Asn Trp Phe Ser 1505 1510 1515 1520 Lys Val Asn Pro Glu Ser Gly Ile Asp Asp Ile Phe Asn Phe Lys Thr 1525 1530 1535 Phe Ala Ser Ser Met Leu Cys Leu Phe Gln Ile Ser Thr Ser Ala Gly 1540 1545 1550 Trp Asp Ser Leu Leu Ser Pro Met Leu Arg Ser Lys Glu Ser Cys Asn 1555 1560 1565 Ser Ser Ser Glu Asn Cys His Leu Pro Gly Ile Ala Thr Ser Tyr Phe 1570 1575 1580 Val Ser Tyr Ile Ile Ile Ser Phe Leu Ile Val Val Asn Met Tyr Ile 1585 1590 1595 1600 Ala Val Ile Leu Glu Asn Phe Asn Thr Ala Thr Glu Glu Ser Glu Asp 1605 1610 1615 Pro Leu Gly Glu Asp Asp Phe Asp Ile Phe Tyr Glu Val Trp Glu Lys 1620 1625 1630 Phe Asp Pro Glu Ala Thr Gln Phe Ile Lys Tyr Ser Ala Leu Ser Asp 1635 1640 1645 Phe Ala Asp Ala Leu Pro Glu Pro Leu Arg Val Ala Lys Pro Asn Lys 1650 1655 1660 Tyr Gln Phe Leu Val Met Asp Leu Pro Met Val Ser Glu Asp Arg Leu 1665 1670 1675 1680 His Cys Met Asp Ile Leu Phe Ala Phe Thr Ala Arg Val Leu Gly Gly 1685 1690 1695 Ser Asp Gly Leu Asp Ser Met Lys Ala Met Met Glu Glu Lys Phe Met 1700 1705 1710 Glu Ala Asn Pro Leu Lys Lys Leu Tyr Glu Pro Ile Val Thr Thr Thr Thr 1715 1720 1725 Lys Arg Lys Glu Glu Glu Arg Gly Ala Ala Ile Ile Gln Lys Ala Phe 1730 1735 1740 Arg Lys Tyr Met Met Lys Val Thr Lys Gly Asp Gln Gly Asp Gln Asn 1745 1750 1755 1760 Asp Leu Glu Asn Gly Pro His Ser Pro Leu Gln Thr Leu Cys Asn Gly 1765 1770 1775 Asp Leu Ser Ser Phe Gly Val Ala Lys Gly Lys Val His Cys Asp 1780 1785 1790 <210> 3 <211> 5728 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (200) .. (4534) <300> <301> Soen-Yong Jeong et al. <302> Identification of a Novel Human Voltage-Gated Sodium Channel α Subunit Gene, SCN12A <303> Biochem. Biophys. Res. Commun. <304> 267 <305> 1 <306> 262-270 <307> 2000 <308> GenBank AF150882 <309> 2000-01-15 <400> 3 tcctgctata cccacagtgg tggtcatctc ttctgatctt cacagccaat cagctcccaa 60 ggcccctgac ctcagctcag cttttgtaga tccttatgac accatccttt aagactggaa 120 tcctagggca ggctgtttta ttcccgcctc ctgaggcctt tctgaggatc tgtggcttgt 180 ctctgtcctg agggtgaag atg gat gac aga tgc tac cca gta atc ttt cca 232 Met Asp Asp Arg Cys Tyr Pro Val Ile Phe Pro 1 5 10 gat gag cgg aat ttc cgc ccc ttc act tcc gac tct ctg gct gca att 280 Asp Glu Arg Asn Phe Arg Pro Phe Thr Ser Asp Ser Leu Ala Ala Ile 15 20 25 gag aag cgg att gcc atc caa aag gag aaa aag aag tct aaa gac cag 328 Glu Lys Arg Ile Ala Ile Gln Lys Glu Lys Lys Lys Ser Lys Asp Gln 30 35 40 aca gga gaa gta ccc cag cct cgg cct cag ctt gac cta aag gcc tcc 376 Thr Gly Glu Val Pro Gln Pro Arg Pro Gln Leu Asp Leu Lys Ala Ser 45 50 55 agg aag ttg ccc aag ctc tat ggc gac att cct cgt gag ctc ata gga 424 Arg Lys Leu Pro Lys Leu Tyr Gly Asp Ile Pro Arg Glu Leu Ile Gly 60 65 70 75 aag cct ctg gaa gac ttg gac cca ttc tac cga aat cat aag aca ttt 472 Lys Pro Leu Glu Asp Leu Asp Pro Phe Tyr Arg Asn His Lys Thr Phe 80 85 90 atg gtg tta aac aga aag agg aca atc tac cgc ttc agt gcc aag cat 520 Met Val Leu Asn Arg Lys Arg Thr Ile Tyr Arg Phe Ser Ala Lys His 95 100 105 gcc ttg ttc att ttt ggg cct ttc aat tca atc aga agt tta gcc att 568 Ala Leu Phe Ile Phe Gly Pro Phe Asn Ser Ile Arg Ser Leu Ala Ile 110 115 120 aga gtc tca gtc cat tca ttg ttc agc atg ttc att atc gg 616 Arg Val Ser Val His Ser Leu Phe Ser Met Phe Ile Ile Gly Thr Val 125 130 135 atc atc aac tgc gtg ttc atg gct aca ggg cct gct aaa aac agc aac 664 Ile Ile Asn Cys Val Phe Met Ala Thr Gly Pro Ala Lys Asn Ser Asn 140 145 150 155 agt aac aat act gac att gca gag tgt gtc ttc act ggg att tat att 712 Ser Asn Asn Thr Asp Ile Ala Glu Cys Val Phe Thr Gly Ile Tyr Ile 160 165 170 ttt gaa gct ttg att aaa ata ttg gca aga ggt ttc att ctg gat gag 760 Phe Glu Ala Leu Ile Lys Ile Leu Ala Arg Gly Phe Ile Leu Asp Glu 175 180 185 ttt tct ttc ctt cga gat cca tgg aac tgg ctg gac tcc att gtc att 808 Arg Asp Pro Trp Asn Trp Leu Asp Ser Ile Val Ile 190 195 200 gga ata gcg att gtg tca tat att cca gga atc acc atc aaa cta ttg 856 Gly Ile Ala Ile Val Ser Tyr Ile Pro Gly Ile Thr Ile Lys Leu Leu 205 210 215 ccc ctg cgt acc ttc cgt gtg ttc aga gct ttg aaa gca att tca gta 904 Pro Leu Arg Thr Phe Arg Val Phe Arg Ala Leu Lys Ala Ile Ser Val 220 225 230 235 gtt tca cgt ctg aag gtc atc gtg ggg gcc ttg tct gtg aag 952 Val Ser Arg Leu Lys Val Ile Val Gly Ala Leu Leu Arg Ser Val Lys 240 245 250 aag ctg gtc aac gtg att atc ctc acc ttc ttt tgc ctc agc atc ttt 1000 Lys Leu Val Asn Val Ile Ile Leu Thr Phe Phe Cys Leu Ser Ile Phe 255 260 265 gcc ctg gta ggt cag cag ctc ttc atg gga agt ctg aac ctg aaa tgc 1048 Ala Leu Val Gly Gln Gln Leu Phe Met Gly Ser Leu Asn Leu Lys Cys 270 275 275 atc tcg ag aaa aat atc agt aac ccg gaa gct tat gac cat 1096 Ile Ser Arg Asp Cys Lys Asn Ile Ser Asn Pro Glu Ala Tyr Asp His 285 290 295 tgc ttt gaa aag aaa gaa aat tca cct gaa ttc aaa atg tgt ggc atc 1 Phe Glu Lys Lys Glu Asn Ser Pro Glu Phe Lys Met Cys Gly Ile 300 305 310 315 tgg atg ggt aac agt gcc tgt tcc ata caa tat gaa tgt aag cac acc 1192 Trp Met Gly Asn Ser Ala Cys Ser Ile Gln Tyr Glu Cys Lys His Thr 320 325 330 aaa att aat cct gac tat aat tat acg aat ttt gac aac ttt ggc tgg 1240 Lys Ile Asn Pro Asp Tyr Asn Tyr Thr Asn Phe Asp Asn Phe Gly Trp 335 340 345 345 tct ttt ctt gcc atg ttc cgg ctg atg acc caa gat tcc tgg gag aag 1288 Ser Phe Leu Ala Met Phe Arg Leu Met Thr Gln Asp Ser Trp Glu Lys 350 355 360 ctt tat caa cag acc ctg cgt act act ggg ctc tac tca gtc ttc ttc 1336 Leu Tyr Gln Gln Thr Leu Arg Thr Thr Gly Leu Tyr Ser Val Phe Phe 365 370 375 ttc att gtg gtc att ttc ctg ggc tcc ttc tac ctg att aac tta acc 1384 Phe Ile Val Val Ile Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Thr 380 385 390 ctg gct gtt gtt acc atg gca tat gag gag cag aac aag aat gta gct 1432 Leu Ala Val Val Thr Met Ala Tyr Glu Glu Gln Asn Lys Asn Val Ala 400 405 410 gca gag ata gag gcc aag gaa aag atg ttt cag gaa gc c cag cag ctg 1480 Ala Glu Ile Glu Ala Lys Glu Lys Met Phe Gln Glu Ala Gln Gln Leu 415 420 425 tta aag gag gaa aag gag gct ctg gtt gcc atg gga att gac aga agt 1528 Leu Lys Glu Glu Lys Glu Ala Ala Met Gly Ile Asp Arg Ser 430 435 440 tca ctt act tcc ctt gaa aca tca tat ttt acc cca aaa aag aga aag 1576 Ser Leu Thr Ser Leu Glu Thr Ser Tyr Phe Thr Pro Lys Lys Arg Lys 445 450 455 ctc ttt ggt aat aag aaa agg aag tcc ttc ttt ttg aga gag tct ggg 1624 Leu Phe Gly Asn Lys Lys Arg Lys Ser Phe Phe Leu Arg Glu Ser Gly 460 465 470 470 475 aaa gac cag cct cct ggg tca gat tct gat gaa gat tgc caaaaa Lys Asp Gln Pro Pro Gly Ser Asp Ser Asp Glu Asp Cys Gln Lys Lys 480 485 490 cca cag ctc cta gag caa acc aaa cga ctg tcc cag aat cta tca ctg 1720 Pro Gln Leu Leu Glu Gln Thr Lys Arg Leu Ser Gln Asn Leu Ser Leu 495 500 505 gac cac ttt gat gag cat gga gat cct ctc caa agg cag aga gca ctg 1768 Asp His Phe Asp Glu His Gly Asp Pro Leu Gln Arg Gln Arg Ala Leu 510 515 520 agt gcc gtc agc atc ctc acc atc acc atg aag gaa caa gaa aaa tca 1816 Ser Ala Val Ser Ile Leu Thr Ile Thr Met Lys Glu Gln Glu Lys Ser 525 530 535 caa gag cct tgt ctc cct tgc gga gaa aac ctg gca tcc aag tac ctc 1864 Gln Glu Pro Cys Pro Cys Gly Glu Asn Leu Ala Ser Lys Tyr Leu 540 545 550 555 gtg tgg aac tgt tgc ccc cag tgg ctg tgc gtt aag aag gtc ctg aga 1912 Val Trp Asn Cys Cys Pro Gln Trp Leu Cys Val Lys Lys Val Leu Arg 560 570 act gtg atg act gac ccg ttt act gag ctg gcc atc acc atc tgc atc 1960 Thr Val Met Thr Asp Pro Phe Thr Glu Leu Ala Ile Thr Ile Cys Ile 575 580 585 atc atc aac act gtc ttc ttg gcc atg gag cat cac aag atg gag gcc 2008 Ile Ile Asn Thr Val Phe Leu Ala Met Glu His His Lys Met Glu Ala 590 595 600 agt ttt gag aag atg ttg aat ata ggg aat ttg gtt ttc act agc att 2056 Ser Phe Glu Lys Met Leu Asn Ile Gly Asn Leu Val Phe Thr Ser Ile 605 610 615 ttt ata gca gaa atg tgc cta aaa atc att gcg ctc gat ccc tac cac 2104 Phe Ile Ala Glu Met Cys Leu Lys Ile Ile Ala Leu Asp Pro Tyr His 620 625 630 630 635 tac ttt cgc cga ggc tgg aac att ttt gac agc att gtt gct ctt ctg 2152 Tyr Phe Arg Arg Gly Trp Asn Ile Phe Asp Ser Ile Val Ala Leu Leu 640 645 agt ttt gca gat gta atg aac tgt gta ctt caa tag caga 2200 Ser Phe Ala Asp Val Met Asn Cys Val Leu Gln Lys Arg Ser Trp Pro 655 660 665 ttc ttg cgt tcc ttc agg gtg ctc agg gtc ttc aag tta gcc aaa tcc 2248 Phe Leu Arg Ser Phe Arg Val Leu Arg Val Phe Lys Ala Lys Ser 670 675 680 tgg cca act ttg aac aca cta att aag ata atc ggc aac tct gtc gga 2296 Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile Gly Asn Ser Val Gly 685 690 695 gcc ctt gga aac ctg act gtg gtg ctg gtc att gtg atc ttt att ttc 2344 Ala Leu Gly Asn Leu Thr Val Val Leu Val Ile Val Ile Phe Ile Phe 700 705 710 715 tca gta gtt ggc atg cag ctt ttt ggc cgt agc ttc aat tcc caa aag 2392 Ser Val Met Gln Leu Phe Gly Arg Ser Phe Asn Ser Gln Lys 720 725 730 agt cca aaa ctc tgt aac ccg aca ggc ccg aca gtc tca tgt tta cgg 2440 Ser Pro Lys Leu Cys Asn Pro Thr Gly Pro Thr Val Ser Cys Leu Arg 735 740 745 cac tgg cac atg ggg gat ttc tgg cac tcc ttc cta gtg gta ttc cgc 2488 His Trp His Met Gly Asp Phe Trp His Ser Phe Leu Val Val Phe Arg 750 755 760 atc ctc tgc ggg gaa tgg atg gaa gaa tgt atg caa gaa 2536 Ile Leu Cys Gly Glu Trp Ile Glu Asn Met Trp Glu Cys Met Gln Glu 765 770 775 gcg aat gca tca tca tca ttg tgt gtt att gtc ttc ata ttg atc acg 2584 Ala Sern As Ser Ser Val Ile Val Phe Ile Leu Ile Thr 780 785 790 795 gtg ata gga aaa ctt gtg gtg ctc aac ctc ttc att gcc tta ctg ctc 2632 Val Ile Gly Lys Leu Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu 800 t 805 810 810 ttt agc aat gag gaa aga aat gga aac tta gaa gga gag gcc 2680 Asn Ser Phe Ser Asn Glu Glu Arg Asn Gly Asn Leu Glu Gly Glu Ala 815 820 825 agg aaa act aaa gtc cag tta gca ctg gat cga ttccg 2728 Arg Lys Thr Lys Val Gln Leu Ala Leu Asp Arg Phe Arg Arg Ala Phe 830 835 840 tgt ttt gtg aga cac act ctt gag cat ttc tgt cac aag tgg tgc agg 2776 Cys Phe Val Arg His Thr Leu Glu His Phe Cys H is Lys Trp Cys Arg 845 850 855 aag caa aac tta cca cag caa aaa gag gtg gca gga ggc tgt gct gca 2824 Lys Gln Asn Leu Pro Gln Gln Lys Glu Val Ala Gly Gly Cys Ala Ala 860 865 870 875 caa agc aaa g att ccc ctg gtc atg gag atg aaa agg ggc tca 2872 Gln Ser Lys Asp Ile Ile Pro Leu Val Met Glu Met Lys Arg Gly Ser 880 885 890 gag acc cag gag gag ctt ggt ata cta acc tct gta cca aag acc ctg 2920 Glu Thr Gln Glu Glu Leu Gly Ile Leu Thr Ser Val Pro Lys Thr Leu 895 900 905 ggc gtc agg cat gat tgg act tgg ttg gca cca ctt gcg gag gag gaa 2968 Gly Val Arg His Asp Trp Thr Trp Leu Ala Pro Leu Ala Glu Glu Glu 910 915 920 gat gac gtt gaa ttt tct ggt gaa gat aat gca cag cgc atc aca caa 3016 Asp Asp Val Glu Phe Ser Gly Glu Asp Asn Ala Gln Arg Ile Thr Gln 925 930 935 cct gag cct gaa caa cag gcc tat gag cag gag aac aag aag 3064 Pro Glu Pro Glu Gln Gln Ala Tyr Glu Leu His Gln Glu Asn Lys Lys 940 945 950 955 ccc acg agc cag aga gtt caa agt gtg gaa att gac atg ttc tct gaa 3112 Pro Thr Ser Gln Arg Va l Gln Ser Val Glu Ile Asp Met Phe Ser Glu 960 965 970 gat gag cct cat ctg acc ata cag gat ccc cga aag aag tct gat gtt 3160 Asp Glu Pro His Leu Thr Ile Gln Asp Pro Arg Lys Lys Ser Asp Val 975 980 985 acc agt ata cta tca gaa tgt agc acc att gat ctt cag gat ggc ttt 3208 Thr Ser Ile Leu Ser Glu Cys Ser Thr Ile Asp Leu Gln Asp Gly Phe 990 995 1000 gga tgg tta cct gag atg gtt ccc aaa aag caa cca gag aga tgt ttg 3256 Gly Trp Leu Pro Glu Met Val Pro Lys Lys Gln Pro Glu Arg Cys Leu 1005 1010 1015 ccc aaa ggc ttt ggt tgc tgc ttt cca tgc tgt agc gtg gac aag aga 3304 Pro Lys Gly Phe Gly Cys Cys Phe Pro Cy Cys Cy Ser Val Asp Lys Arg 1020 1025 1030 1035 aag cct ccc tgg gtc att tgg tgg aac ctg cgg aaa acc tgc tac caa 3352 Lys Pro Pro Trp Val Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Gln 1040 1045 1050 ata gtc aaacacg tgg ttt gag agc ttt att atc ttt gtg att ctg 3400 Ile Val Lys His Ser Trp Phe Glu Ser Phe Ile Ile Phe Val Ile Leu 1055 1060 1065 ctg agc agt ggg gca ctg ata ttt gaa gat gtt cac ctt gag a ac caa 3448 Leu Ser Ser Gly Ala Leu Ile Phe Glu Asp Val His Leu Glu Asn Gln 1070 1075 1080 ccc aaa atc caa gaa tta cta aat tgt act gac att att ttt aca cat 3496 Pro Lys Ile Gle Gln Glu Leu Leu Asn Cys Thr Asp Ile Ile Phe Thr His 1085 1090 1095 att ttt atc ctg gag atg gta cta aaa tgg gta gcc ttc gga ttt gga 3544 Ile Phe Ile Leu Glu Met Val Leu Lys Trp Val Ala Phe Gly Phe Gly 1100 1105 1110 1115 aag tat ttc acc gcc tgg tgc tgc ctt gat ttc atc att gtg att 3592 Lys Tyr Phe Thr Ser Ala Trp Cys Cys Leu Asp Phe Ile Ile Val Ile 1120 1125 1130 gtc tct gtg acc acc ctc att aac tta atg gaa ttg agg tcc ttc Val Thr Thr Leu Ile Asn Leu Met Glu Leu Lys Ser Phe Arg 1135 1140 1145 act cta cga gca ctg agg cct ctt cgt gcg ctg tcc cag ttt gaa gga 3688 Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Gln Phe Glu Gly 1150 1155 1160 atg aag gtg gtg gtc aat gct ctc ata ggt gcc ata cct gcc att ctg 3736 Met Lys Val Val Val Asn Ala Leu Ile Gly Ala Ile Pro Ala Ile Leu 1165 1170 1175 aat gtt ttg ctt gtc tgc ctc att ttc tgg ctc gta ttt tgt att ctg 3784 Asn Val Leu Leu Val Cys Leu Ile Phe Trp Leu Val Phe Cys Ile Leu 1180 1185 1190 1195 gga gta tac ttc ttt tct gga aaa ttt ggg ata tgc ata 38 Gly Val Tyr Phe Phe Ser Gly Lys Phe Gly Lys Cys Ile Asn Gly Thr 1200 1205 1210 gac tca gtt ata aat tat acc atc att aca aat aaa agt caa tgt gaa 3880 Asp Ser Val Ile Asn Tyr Thr Ile Ile Thr Asn Lys Ser Gln Cys Glu 1215 1220 1225 agt ggc aat ttc tct tgg atc aac cag aaa gtc aac ttt gac aat gtg 3928 Ser Gly Asn Phe Ser Trp Ile Asn Gln Lys Val Asn Phe Asp Asn Val 1230 1235 1240 gga aat gct tac ctc gct gct ctc gtg gca aca ttt aag ggc tgg 3976 Gly Asn Ala Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp 1245 1250 1255 atg gat att ata tat gca gct gtt gat tcc aca gag aaa gaa caa cag 4024 Met Asp Ale Ile Ala Val Asp Ser Thr Glu Lys Glu Gln Gln 1260 1265 1270 1275 cca gag ttt gag agc aat tca ctc ggt tac att tac ttc gta gtc ttt 4072 Pro Glu Phe Glu Ser Asn Ser Leu Gly Tyr Ile Tyr P he Val Val Phe 1280 1285 1290 atc atc ttt ggc tca ttc ttc act ctg aat ctc ttc att ggc gtt atc 4120 Ile Ile Phe Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile 1295 1300 1305 att gac aac ttc aac cag aaa aag tta ggt ggc caa gac att 4168 Ile Asp Asn Phe Asn Gln Gln Gln Lys Lys Leu Gly Gly Gln Asp Ile 1310 1315 1320 ttt atg aca gaa gaa cag aag aaa tac tat aat gca atg aaa aaa 4tta Phe Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu 1325 1330 1335 gga tcc aaa aaa cct caa aaa ccc att cca cgg cct ctg aac aaa tgt 4264 Gly Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Cys 1340 1345 1350 1355 caa ggt ctc gtg ttc gac ata gtc aca agc cag atc ttt gac atc atc 4312 Gln Gly Leu Val Phe Asp Ile Val Thr Ser Gln Ile Phe Asp Ile Ile 1360 1365 1370 atc ata agt ctc att atc cta ag atg atg atg gct gaa tca 4360 Ile Ile Ser Leu Ile Ile Leu Asn Met Ile Ser Met Met Ala Glu Ser 1375 1380 1385 tac aac caa ccc aaa gcc atg aaa tcc atc ctt gac cat ctc aac tgg 4408 Tyr Asn Gln Pro Lys Ala Met Lys Ser Ile Leu Asp His Leu Asn Trp 1390 1395 1400 gtc ttt gtg gtc atc ttt acg tta gaa tgt ctc atc aaa atc ttt gct 4456 Val Phe Val Val Ile Phe Thr Leu Glu Cys Leu Ile Lys Ile Phe 1405 1410 1415 ttg agg caa tac tac ttc acc aat ggc tgg aat tta ttt gac tgt gtg 4504 Leu Arg Gln Tyr Tyr Phe Thr Asn Gly Trp Asn Leu Phe Asp Cys Val 1420 1425 1430 1435 gtc gtg ctt ctt tcc att gtt aatcagcagt cagagggact 4554 Val Val Leu Leu Ser Ile Val Ser Lys 1440 1445 ttaagaacca gaagtaagtt tgtaaatctt atcatttttt gaagtttgtt caaactatcc 4614 acaaagcaga aaactgggcc aagtgtactt tctgaaagaa tagacagggg tactaatgcc 4674 attctctact gggaagttgc taggagatag agagtaattt ctgttccctt aactcactac 4734 acaactgaaa tagagttcaa taatcatgca gctaatgtat tcaatggaaa tagacaaaat 4794 taaaatgact cagaagtttt tgtggtggta acctgattta ttcagataat tgagtcaaag 4854 ttctaccttt cctgcgcaac tataagctct atgagcattt tttagtttta aggatttact 4914 gcccatgttt tttcctttgt tcaaaatcca catgggtgcc agatcacaat tattgtgccc 4974 ctgtggcact gctgtgc ccc ctttcaaccc ctatacaagg ccccatggcc cactctagat 5034 tgggctttca agagtatatc attcagacaa tcaaagcttt ttttctattc tatatcccat 5094 gttaacttaa accacagtgt gaactgtgct tgtctcagat tctttgaccc cgcttaccaa 5154 ccagcctcat tctctaatca gtatattttt cctaatcaca cgcagggtta actaaaaggc 5214 ttataaaaga atacaaaaga aaggaggtta ttaccatact ccttcgttat aaaacacaca 5274 tacacacatc aactttaaaa ttaaattctt catggctaag ccataagtgc ttactctggg 5334 tttttgaagt ctggttttaa ttaaactgtt aacttgacat tttataatta tgtaaaactg 5394 tccaggagcc cttctcatag ttcaaaacac aatgaagaca aagaaaactg attttccaaa 5454 tggaagcttt tttgcgatct cagaaagaat gcatcaaaca gatatattgg atcaagatat 5514 tttggttcct aatgctactg cagcattgca gatgtttcag agaacaacaa caaatatgct 5574 acagttcata gtgtgttctg gtttctaagc tatgatgtta atgtagataa agcttatnaa 5634 tagattcttc catgagttta cagttaagta gacctttaat ttttattatt caattccaat 5694 cacaagtctt ggtgaaattg acagcaggaa aaaa 5728 <210> 4 <211> 1444 <212> PRT <213> Homo sapiens <400> 4 Met Asp Asp Arg Cys Tyr Pro Val Ile Phe Pro Asp Glu Arg Asn Phe 1 5 10 15 Arg Pro Phe Thr Ser Asp Ser Leu Ala Ala Ile Glu Lys Arg Ile Ala 20 25 30 Ile Gln Lys Glu Lys Lys Lys Ser Lys Asp Gln Thr Gly Glu Val Pro 35 40 45 Gln Pro Arg Pro Gln Leu Asp Leu Lys Ala Ser Arg Lys Leu Pro Lys 50 55 60 Leu Tyr Gly Asp Ile Pro Arg Glu Leu Ile Gly Lys Pro Leu Glu Asp 65 70 75 80 Leu Asp Pro Phe Tyr Arg Asn His Lys Thr Phe Met Val Leu Asn Arg 85 90 95 Lys Arg Thr Ile Tyr Arg Phe Ser Ala Lys His Ala Leu Phe Ile Phe 100 105 110 Gly Pro Phe Asn Ser Ile Arg Ser Leu Ala Ile Arg Val Ser Val His 115 120 125 Ser Leu Phe Ser Met Phe Ile Ile Gly Thr Val Ile Ile Asn Cys Val 130 135 140 Phe Met Ala Thr Gly Pro Ala Lys Asn Ser Asn Ser Asn Asn Thr Asp 145 150 155 160 Ile Ala Glu Cys Val Phe Thr Gly Ile Tyr Ile Phe Glu Ala Leu Ile 165 170 175 Lys Ile Leu Ala Arg Gly Phe Ile Leu Asp Glu Phe Ser Phe Leu Arg 180 185 190 Asp Pro Trp Asn Trp Leu Asp Ser Ile Val Ile Gly Ile Ala Ile Val 195 200 205 Ser Tyr Ile Pr o Gly Ile Thr Ile Lys Leu Leu Pro Leu Arg Thr Phe 210 215 220 Arg Val Phe Arg Ala Leu Lys Ala Ile Ser Val Val Ser Arg Leu Lys 225 230 235 240 Val Ile Val Gly Ala Leu Leu Arg Ser Val Lys Lys Leu Val Asn Val 245 250 255 Ile Ile Leu Thr Phe Phe Cys Leu Ser Ile Phe Ala Leu Val Gly Gln 260 265 270 Gln Leu Phe Met Gly Ser Leu Asn Leu Lys Cys Ile Ser Arg Asp Cys 275 280 285 285 Lys Asn Ile Ser Asn Pro Glu Ala Tyr Asp His Cys Phe Glu Lys Lys 290 295 300 Glu Asn Ser Pro Glu Phe Lys Met Cys Gly Ile Trp Met Gly Asn Ser 305 310 315 320 Ala Cys Ser Ile Gln Tyr Glu Cys Lys His Thr Lys Ile Asn Pro Asp 325 330 335 Tyr Asn Tyr Thr Asn Phe Asp Asn Phe Gly Trp Ser Phe Leu Ala Met 340 345 350 Phe Arg Leu Met Thr Gln Asp Ser Trp Glu Lys Leu Tyr Gln Gln Thr 355 360 365 Leu Arg Thr Thr Gly Leu Tyr Ser Val Phe Phe Phe Ile Val Val Ile 370 375 380 Phe Leu Gly Ser Phe Tyr Leu Ile Asn Leu Thr Leu Ala Val Val Thr 385 390 395 400 400 Met Ala Tyr Glu Glu Gln Asn Lys Asn Val Ala Ala Glu Ile Glu Ala 405 410 415 Lys Glu Lys M et Phe Gln Glu Ala Gln Gln Leu Leu Lys Glu Glu Lys 420 425 430 Glu Ala Leu Val Ala Met Gly Ile Asp Arg Ser Ser Leu Thr Ser Leu 435 440 445 Glu Thr Ser Tyr Phe Thr Pro Lys Lys Arg Lys Leu Phe Gly Asn Lys 450 455 460 Lys Arg Lys Ser Phe Phe Leu Arg Glu Ser Gly Lys Asp Gln Pro Pro 465 470 475 480 Gly Ser Asp Ser Asp Glu Asp Cys Gln Lys Lys Pro Gln Leu Leu Glu 485 490 495 Gln Thr Lys Arg Leu Ser Gln Asn Leu Ser Leu Asp His Phe Asp Glu 500 505 510 His Gly Asp Pro Leu Gln Arg Gln Arg Ala Leu Ser Ala Val Ser Ile 515 520 525 Leu Thr Ile Thr Met Lys Glu Gln Glu Lys Ser Gln Glu Pro Cys Leu 530 535 540 Pro Cys Gly Glu Asn Leu Ala Ser Lys Tyr Leu Val Trp Asn Cys Cys 545 550 555 560 Pro Gln Trp Leu Cys Val Lys Lys Val Leu Arg Thr Val Met Thr Asp 565 570 575 Pro Phe Thr Glu Leu Ala Ile Thr Ile Cys Ile Ile Ile Asn Thr Val Val 580 585 590 Phe Leu Ala Met Glu His His Lys Met Glu Ala Ser Phe Glu Lys Met 595 600 605 Leu Asn Ile Gly Asn Leu Val Phe Thr Ser Ile Phe Ile Ala Glu Met 610 615 620 Cys Leu Lys Ile I le Ala Leu Asp Pro Tyr His Tyr Phe Arg Arg Gly 625 630 635 640 Trp Asn Ile Phe Asp Ser Ile Val Ala Leu Leu Ser Phe Ala Asp Val 645 650 655 Met Asn Cys Val Leu Gln Lys Arg Ser Trp Pro Phe Leu Arg Ser Phe 660 665 670 Arg Val Leu Arg Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn 675 680 685 Thr Leu Ile Lys Ile Ile Gly Asn Ser Val Gly Ala Leu Gly Asn Leu 690 695 700 Thr Val Val Leu Val Ile Val Ile Phe Ile Phe Ser Val Val Gly Met 705 710 715 715 720 Gln Leu Phe Gly Arg Ser Phe Asn Ser Gln Lys Ser Pro Lys Leu Cys 725 730 735 Asn Pro Thr Gly Pro Thr Val Ser Cys Leu Arg His Trp His Met Gly 740 745 750 Asp Phe Trp His Ser Phe Leu Val Val Phe Arg Ile Leu Cys Gly Glu 755 760 765 Trp Ile Glu Asn Met Trp Glu Cys Met Gln Glu Ala Asn Ala Ser Ser 770 775 780 Ser Leu Cys Val Ile Val Phe Ile Leu Ile Thr Val Ile Gly Lys Leu 785 790 795 800 Val Val Leu Asn Leu Phe Ile Ala Leu Leu Leu Asn Ser Phe Ser Asn 805 810 815 Glu Glu Arg Asn Gly Asn Leu Glu Gly Glu Ala Arg Lys Thr Lys Val 820 825 830 Gln Leu Ala LeuAsp Arg Phe Arg Arg Ala Phe Cys Phe Val Arg His 835 840 845 Thr Leu Glu His Phe Cys His Lys Trp Cys Arg Lys Gln Asn Leu Pro 850 855 860 Gln Gln Lys Glu Val Ala Gly Gly Cys Ala Ala Gln Ser Lys Asp Ile 865 870 875 880 Ile Pro Leu Val Met Glu Met Lys Arg Gly Ser Glu Thr Gln Glu Glu 885 890 895 Leu Gly Ile Leu Thr Ser Val Pro Lys Thr Leu Gly Val Arg His Asp 900 905 910 Trp Thr Trp Leu Ala Pro Leu Ala Glu Glu Glu Asp Asp Val Glu Phe 915 920 925 925 Ser Gly Glu Asp Asn Ala Gln Arg Ile Thr Gln Pro Glu Pro Glu Gln 930 935 940 Gln Ala Tyr Glu Leu His Gln Glu Asn Lys Lys Pro Thr Ser Gln Arg 945 950 955 960 Val Gln Ser Val Glu Ile Asp Met Phe Ser Glu Asp Glu Pro His Leu 965 970 975 Thr Ile Gln Asp Pro Arg Lys Lys Ser Asp Val Thr Ser Ile Leu Ser 980 985 990 Glu Cys Ser Thr Ile Asp Leu Gln Asp Gly Phe Gly Trp Leu Pro Glu 995 1000 1005 Met Val Pro Lys Lys Gln Pro Glu Arg Cys Leu Pro Lys Gly Phe Gly 1010 1015 1020 Cys Cys Phe Pro Cys Cys Ser Val Asp Lys Arg Lys Pro Pro Trp Val 1025 1030 1035 1040 Ile Trp Trp Asn Leu Arg Lys Thr Cys Tyr Gln Ile Val Lys His Ser 1045 1050 1055 Trp Phe Glu Ser Phe Ile Ile Phe Val Ile Leu Leu Ser Ser Gly Ala 1060 1065 1070 Leu Ile Phe Glu Asp Val His Leu Glu Asn Gln Pro Lys Ile Gln Glu 1075 1080 1085 Leu Leu Asn Cys Thr Asp Ile Ile Phe Thr His Ile Phe Ile Leu Glu 1090 1095 1100 Met Val Leu Lys Trp Val Ala Phe Gly Phe Gly Lys Tyr Phe Thr Ser 1105 1110 1115 1120 Ala Trp Cys Cys Leu Asp Phe Ile Ile Val Ile Val Ser Val Thr Thr 1125 1130 1135 Leu Ile Asn Leu Met Glu Leu Lys Ser Phe Arg Thr Leu Arg Ala Leu 1140 1145 1150 Arg Pro Leu Arg Ala Leu Ser Gln Phe Glu Gly Met Lys Val Val Val 1155 1160 1165 Asn Ala Leu Ile Gly Ala Ile Pro Ala Ile Leu Asn Val Leu Leu Val 1170 1175 1180 Cys Leu Ile Phe Trp Leu Val Phe Cys Ile Leu Gly Val Tyr Phe Phe 1185 1190 1195 1200 Ser Gly Lys Phe Gly Lys Cys Ile Asn Gly Thr Asp Ser Val Ile Asn 1205 1210 1215 Tyr Thr Ile Ile Thr Asn Lys Ser Gln Cys Glu Ser Gly Asn Phe Ser 1220 1225 1230 Trp Ile Asn Gln Lys Val Asn Phe Asp Asn Val Gly Asn Ala Tyr Leu 1235 1240 1245 Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile Ile Tyr 1250 1255 1260 Ala Ala Val Asp Ser Thr Glu Lys Glu Gln Gln Pro Glu Phe Glu Ser 1265 1270 1275 1280 Asn Ser Leu Gly Tyr Ile Tyr Phe Val Val Phe Ile Ile Phe Gly Ser 1285 1290 1295 Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn 1300 1305 1310 Gln Gln Gln Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu Glu 1315 1320 1325 Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro 1330 1335 1340 Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Cys Gln Gly Leu Val Phe 1345 1350 1355 1360 Asp Ile Val Thr Ser Gln Ile Phe Asp Ile Ile Ile Ile Ser Leu Ile 1365 1370 1375 Ile Leu Asn Met Ile Ser Met Met Ala Glu Ser Tyr Asn Gln Pro Lys 1380 1385 1390 Ala Met Lys Ser Ile Leu Asp His Leu Asn Trp Val Phe Val Val Ile 1395 1400 1405 Phe Thr Leu Glu Cys Leu Ile Lys Ile Phe Ala Leu Arg Gln Tyr Tyr 1410 1415 1420 Phe Thr Asn Gly Trp Asn Leu Phe Asp Cys Val Val Val Leu Leu Ser 1425 1430 1435 1440 Ile Va l Ser Lys <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 5 atcggcgact ctgtcggagc ccttgga 27 <210> 6 <211> 24 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 6 tgactgtggt cctggtcatt gtga 24 <210> 7 <211> 26 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 7 ttcatkgcrt trtagtaytt cttctg 26 <210> 8 <211> 27 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 8 gttgaarttr tcwatgatga crccaat 27 <210> 9 <211> 29 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 9 tgcggaatac cactaggaag gagtgccag 29 <210> 10 <211> 34 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 10 gagagcaatt cactcggtta catttacttc gtag 34 <210> 11 <211> 36 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 11 ctccctggca tagccacatc ctactttgtc agttac 36 <210> 12 <211> 27 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 12 ccatcctaat acgactcact atagggc 27 <210> 13 <211> 23 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 13 actcactata gggctcgagc ggc 23 <210> 14 <211> 27 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 14 tggcttgtct ctgtcctgag ggtgaag 27 <210> 15 <211> 24 <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 15 ggtaggcgtg gaggtgaggg ctca 24 <210> 16 <211> <212> DNA <213> Artificial Sequence <220> <221> <222> <223> Artificial Sequence: Synthesized Oligonucleotide <400> 16 ctggttctta aagtccctct gactgctga 29

[Brief description of the drawings]

FIG. 1 is a schematic diagram showing a primer set used for PCR amplification of SCN12A, and an EST sequence and a known sequence on which the climber synthesis was based.

FIG. 2 is a schematic diagram of an SCN12A full-length cDNA clone.

FIG. 3. cDNAs of SCN12A and SCN12A-s
FIG. 4 is a schematic diagram showing the relationship between the structure of the above and clones obtained by RT-PCR and RACE.

FIG. 4 shows the result of identification of a chromosomal locus of SCN12A by the FISH method. Metaphase chromosome plate is human chromosome 3p
The hybridization signal of SCN12A is shown on 23-p21.3 (arrow). In the schematic diagram of human chromosome No. 3, the vertical bar indicates the position of SCN12A.

FIG. 5 shows the results of Northern blot analysis in which the expression of SCN12A in each organ was examined. The upper row shows the results of examining various adult tissues, and the lower row shows the results of examining each central nervous system tissue.

FIG. 6 shows the results of in situ hybridization of SCN12A using rat organs. SCN12A
Specifically, a DIG-labeled RNA probe was used. A: hippocampus, B: olfactory bulb, C: low dilation of the cerebellum, D: high dilation of the cerebellum, E: bottom dilation of the spinal cord, F: high dilation of the spinal cord, G: spleen, H: placenta.

FIG. 7: RT-P of SCN12A using human cell lines
This is the result of CR. The cell line is glioblastoma U2
15. Neuroblastoma LAN-5 and SHSY-5
Y. The whole brain, white matter, cerebellum, spinal cord, and DRG (dorsal root ganglion) were also examined.

 ──────────────────────────────────────────────────の Continued on front page F term (reference) 4B024 AA11 BA80 CA04 EA04 GA11 4B064 AG01 AG27 CA19 CC24 DA01 DA13 4H045 AA10 AA11 BA10 CA40 DA76 EA50 FA72 FA74

Claims (6)

[Claims] 1. A human sodium channel SCN12A having the amino acid sequence of SEQ ID NO: 2. 2. A human sodium channel SCN12A-s having the amino acid sequence of SEQ ID NO: 4. 3. A human gene encoding the SCN12A of claim 1 and the SCN12A-s of claim 2, wherein the SCN12A gene is located on the third short arm of human chromosome (3p23-p21.3). 4. The polynucleotide of the human gene according to claim 3, which has the nucleotide sequence of SEQ ID NO: 1. 5. A polynucleotide of the human gene according to claim 3, which has the nucleotide sequence of SEQ ID NO: 3. 6. An antibody against SCN12A of claim 1 or SCN12A-s of claim 2.