CN112342206B - Human PFBC pathogenic gene MYORG with 103 th site mutation and application thereof - Google Patents

Abstract

The invention discloses a new pathogenic gene of hereditary primary familial cerebral calcification, namely MYORG gene, and provides a gene with the sequence shown in SEQ ID NO:4, and further provides a coding protein sequence of the MYORG mutant gene, wherein the coding protein sequence is shown in SEQ ID NO:14, the sequences and the mutation sites thereof can be used as targets for PFBC diagnosis method detection targets and prevention and treatment drug development, and experimental basis and theoretical basis can be provided for PFBC pathogenesis research, clinical diagnosis and treatment methods and drug development. Aiming at the detection of the MYORG gene mutation, the invention develops a corresponding detection kit and a detection method based on PCR amplification and Sanger sequencing, and can simply and effectively detect the MYORG gene mutation.

Description

Human PFBC pathogenic gene MYORG with 103 th site mutation and application thereof

The scheme is a divisional application of patent application 201810552232.6 (application date: 2018, 5, 31, the invention name: primary familial cerebral calcification pathogenic gene MYORG and application thereof).

Technical Field

The invention relates to a human variant gene, in particular to a pathogenic mutant form of a novel pathogenic gene MYORG for primary familial cerebral calcification. The invention also relates to a protein encoded by the MYORG mutant gene, a mutant gene detection kit and a detection method thereof.

Background

Primary familial cerebral calcification (Primary familial brain calcification, PFBC) is a genetic disorder of the nervous system characterized by symmetric calcification of basal ganglia or other brain sites. This disease is also called Fahr disease because it was systematically described by Fahr scholars in 1930. The disease is more than middle-aged and later onset, the disease course is slower, and no obvious sex difference exists. Clinical symptoms are varied and are mainly manifested by movement disorders, mental disorders, cognitive disorders, seizures, dizziness, headache, etc. The light patients have no obvious clinical symptoms, and the heavy patients have personality and behavior changes, and finally cause psychosis or dementia. There may be significant heterogeneity in clinical manifestations between different families or between patients in the same family. Craniocerebral CT is the main means for diagnosing the disease, and can be expressed as symmetrical lump-shaped calcified foci of pallidum, caudate nucleus, dentate nucleus, white matter of brain, thalamus, cortex and the like, and the calcified foci can be connected into a large sheet shape along with the progress of the disease. The pathology of the disease was found to be mainly the deposition of calcium salt particles (with hydroxyapatite as the main component) in the final arterioles and veins, capillaries.

PFBC is sporadic or familial onset, has a high degree of genetic heterogeneity, and is more studied by autosomal dominant inheritance. PFBC is a serious hazard, however, its treatment is mainly symptomatic and cannot be radically cured. The corresponding clinical symptoms are caused by the location and severity of pathological calcification deposits in the brain. However, there is currently no effective drug that can remove or reduce abnormal calcium deposition. Therefore, only the PFBC pathogenic genes are systematically revealed, on the one hand, prenatal diagnosis can be guided to develop prenatal and postnatal care; on the other hand, the early diagnosis of the PFBC patient is realized, and the clinical symptoms of the PFBC patient are further effectively controlled early so as to delay the course of the disease. Therefore, the discovery of pathogenic genes is particularly important, but the analysis of the pathogenic genes and pathogenesis of PFBC is very limited at the present stage.

Beginning in the 90 s of the 20 th century, researchers around the world have conducted extensive genetic studies on PFBC until 2012, the university of chinese martial arts Liu Jingyu teaches cloning the first pathogenic gene of PFBC, SLC20A2. Thereafter, pathogenic genes including mainly PDGFRB, PDGFB, XPR1 and the like were successively discovered and validated. The SLC20A2 gene encodes a sodium-phosphorus co-transporter (PiT 2) protein involved in extracellular phosphorus uptake. The XPR1 gene encodes a protein involved in the excretion of intracellular phosphorus. The PDGFRB gene encodes a platelet-derived growth factor receptor belonging to the class III receptor tyrosine kinase family; the PDGFB gene codes for platelet derived growth factor, and the combination of the PDGFB gene and the PDGFB gene can activate a downstream signal pathway, participate in the development of human pericerebral cells and maintain the integrity of blood brain barrier.

The invention discovers that MYORG gene is the pathogenic gene of autosomal recessive inherited PFBC for the first time, and the specific expression of MYORG in mouse brain is shown in S100 ⁺ Is a cell line of astrocytes. MYORG, also known as KIAA1161 or NET37, located on chromosome 9p13.3 encodes a 714 amino acid protein, a member of the glycosyl hydrolase 31 family. The tail feet of astrocytes together with perivascular cells form neurovascular units, which are just sites of craniocerebral pathological calcium deposition; on the one hand, the mutation of MYORG gene can cause astrocyte dysfunction, influence the connection with perivascular cells, further influence the function of a neuroblood unit and cause calcification deposition; on the other hand, it is possible to cause abnormal glycosylation metabolism of proteins in the brain by affecting the activity of glycosidases, leading to calcification. Unlike the 4 pathogenic genes discovered in the past, MYORG is a recessive pathogenic gene newly disclosed as PFBC, suggesting that a novel gene and cell type are involved in the pathogenesis of PFBC. The method provides a basis for functional analysis and application of MYORG genes, and particularly provides a new molecular target and theoretical basis for exploring, diagnosing, treating and screening PFBC pathogenesis.

Disclosure of Invention

The main purpose of the invention is to reveal a novel pathogenic gene MYORG of PFBC and provide related mutant gene forms thereof; on the basis, a coding protein corresponding to the MYORG mutant gene, a MYORG mutant gene detection kit and a detection method, a carrier, a host cell and a kit containing the MYORG mutant gene are provided, and applications of the mutant gene, the protein, the detection kit, the detection method, the carrier and the cell, especially applications in PFBC are further provided.

The technical scheme adopted for solving the technical problems is as follows:

it is an object of the present invention to provide pathogenic genes associated with autosomal recessive inherited PFBC.

The present invention provides 9 mutant forms of the human MYORG gene, the coding sequences of which are as SED ID NO:2-10, the coding sequence of the MYORG wild type gene of a normal person is shown as SED ID NO: 1. Has the sequence of SEQ ID NO:2-10, and a diploid homozygous form of a MYORG mutant gene of any one of the coding sequences shown in SEQ ID NO: the appearance of either of the two types of diploid heterozygous forms consisting of the MYORG mutant genes of any two coding sequences shown in sequences 2-10 can lead to the occurrence of primary familial cerebral calcification in humans. Reference SED ID NO:1, a MYORG wild-type gene coding sequence shown in the sequence, SED ID NO:2, wherein the mutation of the MYORG mutant gene of the coding sequence shown in the formula 2 occurs at the 225 th site of the wild type gene, and particularly, the mutation of the G base of the wild type gene into the A base is that c.225G > A mutation occurs; SED ID NO:3, wherein the mutation of the MYORG mutant gene of the coding sequence shown in the formula 3 occurs at a 1328 th site of a wild type gene, and particularly, the mutation of a G base of the wild type gene into an A base is that c.1328G > A mutation occurs; SED ID NO:4, wherein the mutation of the MYORG mutant gene of the coding sequence shown in the formula 4 occurs at the 103 th site of the wild type gene, and particularly, the mutation of the A base of the wild type gene into the G base is that c.103A > G mutation occurs; SED ID NO:5, wherein the mutation of the MYORG mutant gene of the coding sequence shown in the formula (1) is generated at a 1321 th site of a wild type gene, and particularly, the mutation of a C base of the wild type gene into a G base is generated by the mutation of c.1321C > G; SED ID NO:6, the mutation of the MYORG mutant gene of the coding sequence shown in the formula 6 occurs at the 695 th site of the wild type gene, and particularly, the mutation of C base of the wild type gene into T base is that the c.695C > T mutation occurs; SED ID NO:7, the mutation of the MYORG mutant gene of the coding sequence shown in the formula 7 occurs at a 607 th site of a wild type gene, and particularly, the mutation of a C base of the wild type gene into a T base is that the c.607C > T mutation occurs; SED ID NO:8, the mutation of the MYORG mutant gene of the coding sequence shown in the formula I occurs at 782 and 783 sites of a wild type gene, and particularly, the mutation of GC base of the wild type gene to TT base is c.782_783GC > TT mutation; SED ID NO:9, wherein the mutation of the MYORG mutant gene of the coding sequence shown in the specification occurs between 348 and 349 sites of the wild type gene, and particularly the c.348-349 insCTGGCCTTCCGC mutation occurs by inserting CTGGCCTTCCGC 12 bases into the site of the wild type gene; SED ID NO:10, wherein mutation of the MYORG mutant gene of the coding sequence shown in the specification occurs in 1092-1097 locus region of the wild type gene, specifically, c.1092-1097 delCTTCGA mutation occurs when 6 base CTTCGA of the wild type gene is deleted.

In view of the disclosure of the MYORG pathogenic gene mutations, the MYORG gene fragments comprising the mutations c.225G > a, c.1328G > a, c.103a > G, c.1321c > G, c.695c > T, c.603 c > T, c.782_783gc > tt, c.348_349 insggcctccgc, and c.1092_1097delCTTCGA can be amplified or synthesized following molecular biological methods and procedures and introduced into vectors to obtain recombinant vectors comprising the mutated gene fragments, which can further be introduced into cells to obtain cells comprising the recombinant vectors, which vectors and cells can be used in research, diagnosis and treatment targets including, but not limited to PFBC mechanisms.

The change of the coding sequence of the gene will cause the change of the protein sequence of the coding protein, and the invention also provides the protein sequence of the coding protein corresponding to the 9 mutant forms of the MYORG gene.

Such as SED ID NO:1, and the protein sequence encoded by the wild MYORG gene is represented by SED ID NO: 11. With SED ID NO:11 as a reference, for SED ID NO:2, and the protein sequence of the coding protein of the MYORG mutant gene is shown as SED ID NO:12, specifically, mutation of a chromogenic amino acid at a 75 th site of a MYORG wild-type gene coding protein to a stop codon, namely p.W75 is mutated; for SED ID NO:3, which encodes a protein sequence of a protein encoded by a MYORG mutant gene, such as SED ID NO:13, specifically, the color amino acid at 443 th site of MYORG wild-type gene coding protein is mutated to a stop codon, i.e. p.W443 is mutated; for SED ID NO:4, and the protein sequence of the coding protein of the MYORG mutant gene is shown as SED ID NO:14, specifically the mutation of the amino acid of the egg at the 35 th site of the MYORG wild-type gene coding protein to valine, namely the p.M35V mutation; for SED ID NO:5, and the protein sequence of the coding protein of the MYORG mutant gene is shown as SED ID NO:15, specifically, the arginine at 441 st position of MYORG wild-type gene encoding protein is mutated into glycine, namely p.R441G mutation occurs; for SED ID NO:6, the coding sequence of the MYORG mutant gene is shown as a protein sequence of the coding protein, such as SED ID NO:16, specifically that serine at 232 th site of MYORG wild type gene coding protein is mutated into leucine, i.e. p.S232L mutation occurs; for SED ID NO:7, which encodes a protein sequence such as SED ID NO:17, specifically, mutation of glutamine at 203 th site of MYORG wild type gene coding protein to stop codon, i.e. p.Q203 is mutated; for SED ID NO:8, and the protein sequence of the coding protein of the MYORG mutant gene is shown as SED ID NO:18, specifically, the mutation of arginine at 261 st site of MYORG wild type gene coding protein to leucine, i.e. the p.R261L mutation; for SED ID NO:9, and the protein sequence of the encoded protein of the MYORG mutant gene is shown as SED ID NO:19, specifically, the position 116 of MYORG wild-type gene coding protein is sequentially inserted with 4 amino acids of leucine, alanine, phenylalanine and arginine, namely, p.116_117insLAFR mutation is generated; for SED ID NO:10, which encodes a protein sequence of a protein encoded by a MYORG mutant gene, such as SED ID NO:20, in particular the phenylalanine and aspartic acid at positions 365 and 366 of the MYORG wild type gene encoding protein are deleted, i.e. the p.365_366delFD mutation is generated. SEQ ID NO:11, and the sequence of SEQ ID NO: the occurrence of any one or more of the MYORG mutant gene-encoded proteins shown in the sequences 12-20 reveals the occurrence of primary familial cerebral calcification.

Furthermore, 9 mutations to the above MYORG gene, SED ID NO:2-10, and a corresponding mutation detection kit. The kit consists of a PCR primer combination designed by the inventor and used for amplifying fragments of a MYORG gene mutation site, preferably a primer combination sequence such as SED ID NO:21-26 comprising SEQ ID NO:21-22, SEQ ID NO:23-24 and SEQ ID NO:25-26 pairs of primers. Wherein, SEQ ID NO:21-22 for amplification of a MYORG gene fragment where the c.103A > G and c.225G > A mutations are located, SEQ ID NO:23-24 for amplification of the MYORG gene fragment where the c.348_349insCTGGCCTTCCGC, c.607C > T, c.695C > T and c.782_783GC > TT mutations are located, SEQ ID NO: the primer pair shown in 25-26 is used for amplifying MYORG gene fragments where c.1092_1097delCTTCGA, c.1321C > G and c.1328G > A mutations are located. In addition, the kit also contains other necessary components of PCR, mainly related reagents such as DNA polymerase, PCR buffer solution, dNTP Mix and the like which can be used for PCR.

Finally, the invention also provides a MYORG gene mutation detection method based on the kit, which mainly comprises the following steps:

(1) Extracting genomic DNA from a biological sample;

(2) PCR amplification of the MYORG gene mutant fragment is carried out by adopting the kit;

(3) Sequencing the PCR product of (2) to determine mutation information of the MYORG gene in the sample.

Wherein the biological sample in step (1) is a body fluid including, but not limited to, blood, serum, amniotic fluid, and lymph. Preferably, the body fluid is blood.

Wherein the primer combination used in the PCR amplification in step (2) is preferably SED ID NO:21-26.

Wherein the mutation information in step (3) is referred to SEQ ID NO:1, and the coding sequence of the MYORG wild type gene.

The invention reveals 9 pathogenic mutations of the novel pathogenic gene MYORG of PFBC and the corresponding coding protein sequences, can be used as a target for PFBC diagnosis method detection and prevention and treatment drug development, and can also provide experimental basis and theoretical basis for PFBC pathogenesis research and clinical diagnosis and treatment methods and drug development. Aiming at the detection of the MYORG gene mutation, the invention develops a corresponding detection kit and a detection method based on PCR amplification and Sanger sequencing, and can simply and effectively detect the MYORG gene mutation.

Drawings

FIGS. 1-6 are, respectively, family diagrams of families A-F and Sanger sequencing plots of the corresponding MYORG gene complex heterozygous or homozygous mutations, wherein the top half is the family diagram and the bottom half is the corresponding Sanger sequencing diagram. Squares in each figure represent men and circles represent women; arrows indicate the first person, with diagonal lines for death, with question marks for non-craniocerebral CT examination of the individual; filled indicates PFBC afflicted individuals and open indicates normal individuals; "-" indicates no mutation detected in the MYORG gene, two lines indicate near wedding, I, II and III represent generations 1, 2 and 3, respectively, and the numbers are member numbers; in each figure, the black boxes are mutant bases, and the same upper and lower bases in the boxes are homozygous mutations. Wherein figure 1 corresponds to family a in example 1, being a MYORG gene c.225g > a (p.w75 x) homozygous mutation, i.e. having SED ID NO: 2; FIG. 2 corresponds to family B, being a compound heterozygous mutation of the MYORG genes c.1328G > A (p.W443) and c.103A > G (p.M35V), i.e.with the SED ID NO:3, 4; FIG. 3 corresponds to family C, being the MYORG genes c.225G > A (p.W75) and c.1321C > G (p.R441G) complex heterozygous mutations, i.e. with SED ID NO:2, 5; FIG. 4 corresponds to family D, being a homozygous mutation of the MYORG gene c.695C > T (p.S232L), i.e. having the SED ID NO: 6; FIG. 5 corresponds to family E, the MYORG genes c.603C > T (p.Q203) and c.782_783GC > TT (p.R261L) are complex heterozygous mutations, i.e. with SED ID NO:7, 8; FIG. 6 corresponds to family F, the MYORG genes c.348_349insCTGGCCTTCCGC (p.116_117 insLAFR) and c.1092_1097delCTTCGA (p.365_366 delFD) complex heterozygous mutations, i.e.with SED ID NO:9, 10.

FIG. 7 is a CT pan-scan of the cranium of PFBC patients V7 (left half) and V9 (right half) in example 1.

Detailed Description

The invention is further illustrated below in conjunction with specific examples.

The inventors revealed for the first time the pathogenic mutant form of the pathogenic gene MYORG of autosomal recessive inherited PFBC by whole exon sequencing of two autosomal recessive inherited PFBC families a and B and Sanger sequencing of the other 11 PFBC families. The experimental procedures, which are not specified in the following examples, are generally carried out under conventional conditions or under conditions recommended by the manufacturer.

EXAMPLE 1 preliminary identification of pathogenic genes of PFBC families A and B Using full exon sequencing

Based on detailed medical history and family history investigation and standardized physical examination of the nervous system and related auxiliary examination information, 2 PFBC family samples were collected for pathogenic gene detection and analysis.

(1) PFBC family A and B sample information

The inventors first began with 2 autosomal recessive inherited PFBC families (families a and B) and examined PFBC patients in these 2 families did not detect the relevant mutations of the known pathogenic genes (SLC 20A2, PDGFRB, PDGFB and XPR 1), suggesting that new PFBC pathogenic genes are possible. Wherein family A contains 2 PFBC patients (numbers V7 and V9, V9 are forerunner), all presenting as globus pallidus and cerebellum large area calcification foci (FIG. 7), dysarthria, ataxia, and parents are close-related weddings. There were also 2 PFBC patients in family B, with the distribution of cranial calcification lesions and severity of both precursor (III 4) and its siblings (III 6) being highly similar to family a, and the craniocerebral CT of their parents (II 3, II 4) were negative.

(2) Whole exome sequencing analysis

2 PFBC patients in family a (V7, V9) and unaffected brothers (V5), 2 PFBC patients in family B (III 4, III 6) and unaffected sister (III 5) were selected for whole exon sequencing analysis. The main flow is as follows: 10mL of peripheral blood was withdrawn, genomic DNA was extracted therefrom using QIAamp DNA Blood Mini Kit (Kaijia, germany), seqCap EZ Exome Kit V3 (Roche, USA) was subjected to whole-Exome capture, and the resultant was sequenced on a Hiseq 3000 second-generation sequencer (Illumina, USA), and gene mutation information was obtained in comparison with human reference genome GRCh 38. Sequencing results showed that more than 70% of reads were located in the target capture region, more than 99.7% of the target region of interest was covered and the average depth of coverage was 82X.

(3) Pathogenic gene screening

Based on genetic pattern analysis, gene screening is performed in a putative autosomal recessive genetic manner, giving preference to homozygous mutations. The total exon sequencing data of the family a samples were screened for 5 genes with co-segregation and mutations thereof, respectively, c.607g < C (p.v203l) of DDX1 gene, c.c506C < T (p.t169i) of raggap 1L gene, c.225g > a (p.w75 x) of MYORG gene, 3058C > g (p.q 1020e) of zchc 6 gene, and c.2971c < T (p.r991w) of TMEM2 gene. Combining the sequencing result of the whole exome of the family B sample, preferentially screening the genes sharing the compound heterozygous mutation or homozygous mutation of two patients but not existing in the control, and finding that the compound heterozygous mutation exists in the MYORG gene of the family B sample: one is a truncation mutation c.1328g > a, p.w443, the coding sequence of which is as set forth in SEQ ID NO:3 is shown in the figure; and the second is missense mutation c.103A > G, namely p.M35V, and the coding sequence is shown as SEQ ID NO:4 is shown in the figure; the wild type sequence of MYORG Gene is shown in NM_020702 sequence of NCBI Gene57462, and the coding sequence is shown in SEQ ID NO: 1.

EXAMPLE 2Sanger sequencing to verify MYORG pathogenic Gene mutations in PFBC families A and B

Since only MYORG is a gene shared in both families (families A and B), the inventors amplified the gene fragment where the mutation was located using the kit and method proposed in the present invention (see example 6 for specific kits and methods), and further performed MYORG gene mutation verification and detection on family A using Sanger sequencing for 12 related members (PFBC patients: V7, V9, non-PFBC members: IV6, IV7, V1, V2, V3, V4, V5, V6, VI1, VI 2). Two patients in family a were found to indeed contain a c.225g > a homozygous mutation for the MYORG gene, whereas none of the remaining 10 normal persons in the family were found to carry the homozygous mutation at this site, indicating that the MYORG gene mutation was coseparated from the disease phenotype in this family. The found mutation of the 225 th base G of the MYORG gene into A, namely c.225G > A, is a new mutation of the gene, so that the 75 th tryptophan of a protein sequence (SEQ ID NO: 12) coded by the MYORG gene is mutated into a stop codon (p.W75 x), and the coding sequence of the mutated gene is shown as SEQ ID NO:2 (fig. 1). In family B, two complex heterozygous MYORG gene mutations also present a family co-segregation phenomenon. Two new mutations found in family B, namely, mutation of 1328 th base G of MYORG gene coding sequence to A, namely, mutation of c.1328G > A (mutation gene coding sequence is shown as SEQ ID NO: 3), mutation of 443 th color amino acid of protein sequence (SEQ ID NO: 13) coded by the gene to stop codon (p.W443), mutation of 103 th base A to G (mutation gene coding sequence is shown as SEQ ID NO: 4), mutation of c.103A > G (mutation gene coding sequence is shown as SEQ ID NO: 14), mutation of 35 th amino acid of protein sequence (SEQ ID NO: 14) coded by the MYORG gene to valine (p.M35V) (FIG. 2).

Example 3 screening for MYORG pathogenic Gene mutations in other PFBC pedigrees

Since only MYORG is a gene shared in families a and B, we further extended family sample validation, mutation screening of the MYORG full coding region was performed on PFBC families with 8 autosomal recessive inheritance and precursors of 3 families with unknown genetic patterns. According to the wild type gene coding sequence SED ID NO of MYORG: 1, designing a primer pair SED ID NO:21-28 and Sanger sequencing was performed. Wherein SED ID NO:27-28 are identical to the SED ID NO:21-26 (see example 6 for specific kits and methods). Wherein, the precursor of the C, D, E, F four families contains MYORG gene compound heterozygous mutation or homozygous mutation. Two complex heterozygous mutations of the MYORG gene are found in the C family, namely c.225G > A (homologous family A), and c.1321C > G, namely 1321 base C, is mutated into G, so that the 441 st arginine of a protein sequence (SEQ ID NO: 15) encoded by the MYORG gene is mutated into glycine (p.R441G), and the coding sequence of the mutated gene is shown as SEQ ID NO:5 (fig. 3). 1 homozygous mutation of the MYORG gene is found in the D family, the mutation of the base C at 695 is T, namely c.695C > T, so that the 232 th serine of a protein sequence (SEQ ID NO: 16) encoded by the MYORG gene is mutated into leucine (p.S232L), and the encoding sequence of the mutated gene is as shown in SEQ ID NO:6 (fig. 4). Two compound heterozygous mutations of the MYORG gene are found in an E family, one is that the mutation is carried out at 607 th base C to T, namely c.607C > T, so that the 203 th glutamine of a protein sequence (SEQ ID NO: 17) encoded by the MYORG gene is mutated to a stop codon (p.Q203 x), and the encoding sequence of the mutated gene is shown as SEQ ID NO: shown at 7. The other is that GC at 782 and 783 bases is mutated into TT, namely c.782_783GC > TT, so that 261 st arginine of a protein sequence (SEQ ID NO: 18) coded by MYORG genes is mutated into leucine (p.R261L), and the coding sequence of the mutated genes is as shown in SEQ ID NO:8 (fig. 5). Two compound heterozygous mutations of the MYORG gene are found in the F family, one is that 12 bases CTGGCCTTCCGC, namely c.348_349insCTGGCCTTCCGC, are inserted between 348 and 349 sites, so that 4 amino acids (leucine, alanine, phenylalanine and arginine, p.116_117 insLAFR) are inserted between 116 and 117 amino acid sites of a protein sequence (SEQ ID NO: 19) encoded by the MYORG gene, and the mutant gene has a coding sequence as shown in SEQ ID NO: shown at 9. The other is that the CTTCGA with 6 bases is deleted at 1092-1097, namely c.1092-1097 delCTTCGA, so that the 365 th and 366 th positions of a protein sequence (SEQ ID NO: 20) coded by a MYORG gene are deleted with phenylalanine and aspartic acid (p.365-366 delFD), and the specific sequence of a mutant gene is shown in SEQ ID NO:10 (fig. 6). Thus, 9 MYORG gene mutations were detected in 6 PFBC family (family a-F) samples altogether (see table 1). The positions and rs ID in table 1 were obtained with reference to Genome Reference Consortium human genome build (GRCh 38), and gnomAD data was derived from the genomic mutation frequency database.

Mutation sites of MYORG genes found in Table 1

Example 4 Using linkage analysis to further demonstrate that pathogenic gene MYORG mutations are completely cosegregated in 6 families with disease phenotypes

All family members of 6 families (families a-F) in the above examples were tested for mutation sites above the MYORG gene by sanger sequencing, reference (Kruglyak, l., daly, m.j., reeve-Daly, m.p., and Lander, e.s., parametric and nonparametric linkage analysis: a unified multipoint appreach.am J Hum Genet 58,1996,1347-1363.), and by a putative autosomal recessive genetic approach, the allele frequency of the disease was 0.01, dominant homozygous normal allele, and heterozygous, homozygous disease allele homozygote, 0.001, 0.999, and 0.999, respectively. The MYORG gene mutation and the calcification phenotype show complete co-segregation phenomenon, and the probability logarithmic ratio is 4.91. Thus, it was determined that the MYORG gene is a causative gene of autosomal recessive PFBC.

Example 5 by analysis of these 9 MYORG mutation sites, it was further clarified that the pathogenic gene MYORG shares 9 MYORG gene mutation sites in 6 PFBC families, of which 3 are nonsense mutations (p.w75, p.q203 and p.w443), 2 are insertion or deletion mutations (p.116_ 117insLAFR and p.365_366delFD), and the remaining 4 are highly conserved missense mutations (p.M35V, p.S232L, p.R261L and p.r441g). None of these mutations were present in the 1000 genome database, the NHLBI exon sequencing database, nor in the genomic mutation frequency database (gnomAD) or at very low frequencies (table 1). The inventors simultaneously took 1000 healthy human control blood samples of non-PFBC, and by referring to the method of example 6, sanger sequencing of all coding regions of MYORG gene was performed, and none of the above 9 mutation sites were found, i.e. none of nonsense mutations, compound heterozygous or homozygous mutations were found, and the pathogenicity of these mutation sites was further clarified.

Example 6 MYORG pathogenic Gene test

Based on the 9 pathogenic mutation gene forms of the PFBC pathogenic genes MYORG disclosed by the invention, the invention further provides a detection kit and a detection method for the MYORG gene mutation, and the detection kit and the detection method are applied to the detection of the MYORG gene mutation of individuals in 6 PFBC families, and specifically comprise the following steps:

(1) Specimen collection and processing

The PFBC family a-F patients and related members were subjected to detailed medical history and family history investigation, and standardized neurological physical examination and related auxiliary examination information, and 5ml of elbow venous blood (sodium citrate anticoagulation) was extracted under the condition that they signed informed consent, and genomic DNA was extracted.

(2) MYORG mutant Gene fragment amplification and sequencing

And (3) using the genome DNA in the step (1) as a template, and carrying out PCR in vitro amplification on the MYORG gene fragment by adopting the kit. The main components of the amplification kit of the invention comprise rTaq polymerase (TaKaRa, commercial code DR 100A) of Japanese Takara company, and 10 XPCR buffer, dNTP Mix and primer combination SED ID NO matched with the rTaq polymerase: 21-26 (primers were synthesized by Shanghai Biotechnology Co., ltd.). The amplification system is shown in Table 2, wherein the selection of the primer pairs F and R is shown in Table 3, and the amplification conditions are as follows: pre-denaturation at 94℃for 3min;30 cycles: denaturation at 94℃for 30s, annealing at 60℃for 30s, extension at 72℃for 30s; finally, the extension is carried out for 5min at 72 ℃.

TABLE 2 amplification System

Numbering device	The components	Additive amount/. Mu.L
			1	10×PCR buffer(Mg 2+ )	2.5
2	dNTP Mix	0.5
			3	Primer F(50μM)	0.25
4	Primer R(50μM)	0.25
			5	rTaq DNA polymerase	0.25
6	Genomic DNA template (30 ng/. Mu.L)	1.0
			7	Sterilizing ddH 2 O	Complement to 20

(3) MYORG gene mutation analysis

And (3) sending the PCR product in the step (2) to Shanghai Biotechnology engineering Co., ltd for Sanger sequencing, and analyzing the sequencing result to find out mutation sites.

TABLE 3 amplification primer pairs

The results (Table 3) show that all 6 families A-F contain complex heterozygous or homozygous mutations. All patients of family a had SED ID NO:2, all patients of family B had the mutation indicated by SED ID NO:3 and 4, all patients of family C had SED ID NO:2 and 5, all patients of family D had SED ID NO:6, all patients of family E had the mutation set forth in SED ID NO:7 and 8, all patients of family F had SED ID NO:9 and 10. In each family, normal individuals and 1000 normal controls (example 5) were not mutated as described above. The method provided by the invention can be used for accurately detecting and analyzing autosomal recessive genetic PFBC.

While the invention has been described in terms of preferred embodiments, various modifications and alterations will occur to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Sequence listing

<110> affiliated first Hospital of university of Fujian medical science

Shanghai Institute of life sciences, Chinese Academy of Sciences

<120> human PFBC pathogenic gene MYORG with mutation at 103 rd site and application thereof

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<170> SIPOSequenceListing 1.0

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<213> Homo sapiens (Homo sapiens)

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gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

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tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 2

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 2

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctgatgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 3

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 3

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactagt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 4

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 4

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctgtgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 5

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 5

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

ggcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 6

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 6

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tctttgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 7

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 7

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggctagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 8

<211> 2145

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 8

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

ctttaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg acttcgatga ggtcaaattc cccaacgcca gcgacatgtt ccgccgcctg 1140

cgcgacgccg gcttccgcgt cacgctctgg gtgcaccctt ttgtcaacta caactcgtcg 1200

cgcttcggcg agggcgtgga gcgcgagctg ttcgtgcgcg aacccacggg ccggttacct 1260

gcgctggtgc gctggtggaa cggcatcggc gcggtgctag acttcacgca cccaaaggcc 1320

cgcgactggt tccagggaca cctgcggcgg ctgcgctctc gctactccgt ggcttccttc 1380

aagttcgacg cgggcgaggt cagctacctg ccgcgggact tcagcaccta ccggccgctg 1440

ccggacccca gcgtctggag ccggcgctac actgagatgg cgctgccctt cttctcgctg 1500

gcggaggtgc gcgtaggcta ccagtcacag aacatctcct gcttcttccg cctggtggat 1560

cgcgactctg tgtggggcta cgacctgggg ttgcgctcac tcatccccgc ggtgctcacc 1620

gtcagcatgc tgggctaccc attcatccta cccgatatgg tgggcggcaa cgccgtgccc 1680

cagcggacag ccggcggcga tgtgcccgag cgcgagctct acattcgctg gctggaagtg 1740

gccgccttta tgccggccat gcagttctct atcccgccct ggcgctacga cgcggaagtg 1800

gtggccatcg cgcagaagtt cgccgccctg cgggcctcgc ttgtggcacc gctgttgctt 1860

gagctggcgg gcgaggtcac cgacacgggt gaccctatcg tgcgccccct ttggtggatt 1920

gcgcccggcg acgagacagc tcaccgtatc gactcgcagt tccttattgg ggacacgctg 1980

cttgtggccc cggtgctgga gccaggcaag caggagcgcg acgtctattt gcccgccggc 2040

aagtggcgca gctacaaggg tgagcttttc gacaagacgc cggtgctgct caccgattac 2100

ccggtcgacc tggatgagat cgcctacttt acctgggcgt cctga 2145

<210> 9

<211> 2157

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 9

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgcct ggccttccgc 360

tccggcgcgc tggaccttga ctcctgcagc cgcgatggcg ccctgctggg ctgctcgctc 420

acggccgacg ggctgccgct gcacttcttc atccagactg tgcggcccaa ggacacggtc 480

atgtgctacc gcgtgcgctg ggaggaggca gcgccgggcc gggccgtgga gcacgccatg 540

ttcttgggcg acgcggcggc ccactggtat ggtggcgccg agatgaggac gcaacactgg 600

cccatccgcc tggatggcca gcaggagccc cagccgttcg tcaccagcga tgtctactcc 660

tccgacgccg cgtttggggg catcctcgag cgctactggc tatcttcgcg cgcggccgcc 720

atcaaagtca atgactcagt gcccttccac ctgggctgga acagcacgga gcgctcgctg 780

cggcttcagg cgcgctacca cgacacgccc tacaagccac ccgccggccg cgccgcagcg 840

ccagagctga gctaccgagt gtgcgtgggc tcagacgtca cctccatcca caagtacatg 900

gtgcgtcgct acttcaacaa gccgtcaagg gtgccagcac ccgaggcctt ccgagacccc 960

atttggtcca catgggcgct gtacgggcgc gccgtggacc aggacaaggt gctgcgtttt 1020

gcccaacaga tccgcctgca ccacttcaac agcagccacc tggaaatcga cgacatgtac 1080

acacctgctt atggcgactt cgacttcgat gaggtcaaat tccccaacgc cagcgacatg 1140

ttccgccgcc tgcgcgacgc cggcttccgc gtcacgctct gggtgcaccc ttttgtcaac 1200

tacaactcgt cgcgcttcgg cgagggcgtg gagcgcgagc tgttcgtgcg cgaacccacg 1260

ggccggttac ctgcgctggt gcgctggtgg aacggcatcg gcgcggtgct agacttcacg 1320

cacccaaagg cccgcgactg gttccaggga cacctgcggc ggctgcgctc tcgctactcc 1380

gtggcttcct tcaagttcga cgcgggcgag gtcagctacc tgccgcggga cttcagcacc 1440

taccggccgc tgccggaccc cagcgtctgg agccggcgct acactgagat ggcgctgccc 1500

ttcttctcgc tggcggaggt gcgcgtaggc taccagtcac agaacatctc ctgcttcttc 1560

cgcctggtgg atcgcgactc tgtgtggggc tacgacctgg ggttgcgctc actcatcccc 1620

gcggtgctca ccgtcagcat gctgggctac ccattcatcc tacccgatat ggtgggcggc 1680

aacgccgtgc cccagcggac agccggcggc gatgtgcccg agcgcgagct ctacattcgc 1740

tggctggaag tggccgcctt tatgccggcc atgcagttct ctatcccgcc ctggcgctac 1800

gacgcggaag tggtggccat cgcgcagaag ttcgccgccc tgcgggcctc gcttgtggca 1860

ccgctgttgc ttgagctggc gggcgaggtc accgacacgg gtgaccctat cgtgcgcccc 1920

ctttggtgga ttgcgcccgg cgacgagaca gctcaccgta tcgactcgca gttccttatt 1980

ggggacacgc tgcttgtggc cccggtgctg gagccaggca agcaggagcg cgacgtctat 2040

ttgcccgccg gcaagtggcg cagctacaag ggtgagcttt tcgacaagac gccggtgctg 2100

ctcaccgatt acccggtcga cctggatgag atcgcctact ttacctgggc gtcctga 2157

<210> 10

<211> 2139

<212> DNA

<213> Homo sapiens (Homo sapiens)

<400> 10

atgctccaga accctcagga gaagagccag gcctaccccc gccgccgccg gcctggctgc 60

tacgcatacc gtcagaaccc cgaggccatc gcagccgcag ctatgtacac cttcctgccc 120

gacaacttct cacctgccaa gcccaagcct tccaaagacc tgaagccgct gctgggctcc 180

gcggttctgg ggctgctgct tgtgctggcc gcggtggtgg cctggtgcta ctacagcgtc 240

tccctacgca aggcggagcg acttcgcgcg gagctgctgg acctgaaagc tggcggcttc 300

tccatccgca atcagaaggg agagcaggtc ttccgcctgg ccttccgctc cggcgcgctg 360

gaccttgact cctgcagccg cgatggcgcc ctgctgggct gctcgctcac ggccgacggg 420

ctgccgctgc acttcttcat ccagactgtg cggcccaagg acacggtcat gtgctaccgc 480

gtgcgctggg aggaggcagc gccgggccgg gccgtggagc acgccatgtt cttgggcgac 540

gcggcggccc actggtatgg tggcgccgag atgaggacgc aacactggcc catccgcctg 600

gatggccagc aggagcccca gccgttcgtc accagcgatg tctactcctc cgacgccgcg 660

tttgggggca tcctcgagcg ctactggcta tcttcgcgcg cggccgccat caaagtcaat 720

gactcagtgc ccttccacct gggctggaac agcacggagc gctcgctgcg gcttcaggcg 780

cgctaccacg acacgcccta caagccaccc gccggccgcg ccgcagcgcc agagctgagc 840

taccgagtgt gcgtgggctc agacgtcacc tccatccaca agtacatggt gcgtcgctac 900

ttcaacaagc cgtcaagggt gccagcaccc gaggccttcc gagaccccat ttggtccaca 960

tgggcgctgt acgggcgcgc cgtggaccag gacaaggtgc tgcgttttgc ccaacagatc 1020

cgcctgcacc acttcaacag cagccacctg gaaatcgacg acatgtacac acctgcttat 1080

ggcgacttcg atgaggtcaa attccccaac gccagcgaca tgttccgccg cctgcgcgac 1140

gccggcttcc gcgtcacgct ctgggtgcac ccttttgtca actacaactc gtcgcgcttc 1200

ggcgagggcg tggagcgcga gctgttcgtg cgcgaaccca cgggccggtt acctgcgctg 1260

gtgcgctggt ggaacggcat cggcgcggtg ctagacttca cgcacccaaa ggcccgcgac 1320

tggttccagg gacacctgcg gcggctgcgc tctcgctact ccgtggcttc cttcaagttc 1380

gacgcgggcg aggtcagcta cctgccgcgg gacttcagca cctaccggcc gctgccggac 1440

cccagcgtct ggagccggcg ctacactgag atggcgctgc ccttcttctc gctggcggag 1500

gtgcgcgtag gctaccagtc acagaacatc tcctgcttct tccgcctggt ggatcgcgac 1560

tctgtgtggg gctacgacct ggggttgcgc tcactcatcc ccgcggtgct caccgtcagc 1620

atgctgggct acccattcat cctacccgat atggtgggcg gcaacgccgt gccccagcgg 1680

acagccggcg gcgatgtgcc cgagcgcgag ctctacattc gctggctgga agtggccgcc 1740

tttatgccgg ccatgcagtt ctctatcccg ccctggcgct acgacgcgga agtggtggcc 1800

atcgcgcaga agttcgccgc cctgcgggcc tcgcttgtgg caccgctgtt gcttgagctg 1860

gcgggcgagg tcaccgacac gggtgaccct atcgtgcgcc ccctttggtg gattgcgccc 1920

ggcgacgaga cagctcaccg tatcgactcg cagttcctta ttggggacac gctgcttgtg 1980

gccccggtgc tggagccagg caagcaggag cgcgacgtct atttgcccgc cggcaagtgg 2040

cgcagctaca agggtgagct tttcgacaag acgccggtgc tgctcaccga ttacccggtc 2100

gacctggatg agatcgccta ctttacctgg gcgtcctga 2139

<210> 11

<211> 714

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 11

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Arg Asp Trp Phe Gln Gly His Leu

435 440 445

Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala

450 455 460

Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu

465 470 475 480

Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro

485 490 495

Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile

500 505 510

Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp

515 520 525

Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu

530 535 540

Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro

545 550 555 560

Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg

565 570 575

Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro

580 585 590

Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala

595 600 605

Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly

610 615 620

Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile

625 630 635 640

Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile

645 650 655

Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu

660 665 670

Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu

675 680 685

Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu

690 695 700

Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 12

<211> 74

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 12

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala

65 70

<210> 13

<211> 442

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 13

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Arg Asp

435 440

<210> 14

<211> 714

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 14

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Val Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Arg Asp Trp Phe Gln Gly His Leu

435 440 445

Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala

450 455 460

Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu

465 470 475 480

Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro

485 490 495

Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile

500 505 510

Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp

515 520 525

Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu

530 535 540

Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro

545 550 555 560

Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg

565 570 575

Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro

580 585 590

Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala

595 600 605

Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly

610 615 620

Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile

625 630 635 640

Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile

645 650 655

Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu

660 665 670

Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu

675 680 685

Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu

690 695 700

Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 15

<211> 714

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 15

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Gly Asp Trp Phe Gln Gly His Leu

435 440 445

Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala

450 455 460

Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu

465 470 475 480

Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro

485 490 495

Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile

500 505 510

Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp

515 520 525

Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu

530 535 540

Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro

545 550 555 560

Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg

565 570 575

Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro

580 585 590

Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala

595 600 605

Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly

610 615 620

Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile

625 630 635 640

Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile

645 650 655

Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu

660 665 670

Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu

675 680 685

Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu

690 695 700

Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 16

<211> 714

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 16

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Leu Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Arg Asp Trp Phe Gln Gly His Leu

435 440 445

Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala

450 455 460

Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu

465 470 475 480

Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro

485 490 495

Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile

500 505 510

Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp

515 520 525

Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu

530 535 540

Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro

545 550 555 560

Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg

565 570 575

Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro

580 585 590

Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala

595 600 605

Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly

610 615 620

Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile

625 630 635 640

Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile

645 650 655

Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu

660 665 670

Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu

675 680 685

Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu

690 695 700

Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 17

<211> 202

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 17

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly

195 200

<210> 18

<211> 714

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 18

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Leu Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Phe Asp Glu Val

355 360 365

Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly

370 375 380

Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser

385 390 395 400

Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr

405 410 415

Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val

420 425 430

Leu Asp Phe Thr His Pro Lys Ala Arg Asp Trp Phe Gln Gly His Leu

435 440 445

Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala

450 455 460

Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu

465 470 475 480

Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro

485 490 495

Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile

500 505 510

Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp

515 520 525

Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu

530 535 540

Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro

545 550 555 560

Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg

565 570 575

Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro

580 585 590

Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala

595 600 605

Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly

610 615 620

Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile

625 630 635 640

Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile

645 650 655

Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu

660 665 670

Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu

675 680 685

Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu

690 695 700

Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 19

<211> 718

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 19

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser Leu Ala Phe Arg

340 345 350

His Leu Glu Ile Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp

355 360 365

Phe Asp Glu Val Lys Phe Pro Asn Ala Ser Asp Met Phe Arg Arg Leu

370 375 380

Arg Asp Ala Gly Phe Arg Val Thr Leu Trp Val His Pro Phe Val Asn

385 390 395 400

Tyr Asn Ser Ser Arg Phe Gly Glu Gly Val Glu Arg Glu Leu Phe Val

405 410 415

Arg Glu Pro Thr Gly Arg Leu Pro Ala Leu Val Arg Trp Trp Asn Gly

420 425 430

Ile Gly Ala Val Leu Asp Phe Thr His Pro Lys Ala Arg Asp Trp Phe

435 440 445

Gln Gly His Leu Arg Arg Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe

450 455 460

Lys Phe Asp Ala Gly Glu Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr

465 470 475 480

Tyr Arg Pro Leu Pro Asp Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu

485 490 495

Met Ala Leu Pro Phe Phe Ser Leu Ala Glu Val Arg Val Gly Tyr Gln

500 505 510

Ser Gln Asn Ile Ser Cys Phe Phe Arg Leu Val Asp Arg Asp Ser Val

515 520 525

Trp Gly Tyr Asp Leu Gly Leu Arg Ser Leu Ile Pro Ala Val Leu Thr

530 535 540

Val Ser Met Leu Gly Tyr Pro Phe Ile Leu Pro Asp Met Val Gly Gly

545 550 555 560

Asn Ala Val Pro Gln Arg Thr Ala Gly Gly Asp Val Pro Glu Arg Glu

565 570 575

Leu Tyr Ile Arg Trp Leu Glu Val Ala Ala Phe Met Pro Ala Met Gln

580 585 590

Phe Ser Ile Pro Pro Trp Arg Tyr Asp Ala Glu Val Val Ala Ile Ala

595 600 605

Gln Lys Phe Ala Ala Leu Arg Ala Ser Leu Val Ala Pro Leu Leu Leu

610 615 620

Glu Leu Ala Gly Glu Val Thr Asp Thr Gly Asp Pro Ile Val Arg Pro

625 630 635 640

Leu Trp Trp Ile Ala Pro Gly Asp Glu Thr Ala His Arg Ile Asp Ser

645 650 655

Gln Phe Leu Ile Gly Asp Thr Leu Leu Val Ala Pro Val Leu Glu Pro

660 665 670

Gly Lys Gln Glu Arg Asp Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser

675 680 685

Tyr Lys Gly Glu Leu Phe Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr

690 695 700

Pro Val Asp Leu Asp Glu Ile Ala Tyr Phe Thr Trp Ala Ser

705 710 715

<210> 20

<211> 712

<212> PRT

<213> Homo sapiens (Homo sapiens)

<400> 20

Met Leu Gln Asn Pro Gln Glu Lys Ser Gln Ala Tyr Pro Arg Arg Arg

1 5 10 15

Arg Pro Gly Cys Tyr Ala Tyr Arg Gln Asn Pro Glu Ala Ile Ala Ala

20 25 30

Ala Ala Met Tyr Thr Phe Leu Pro Asp Asn Phe Ser Pro Ala Lys Pro

35 40 45

Lys Pro Ser Lys Asp Leu Lys Pro Leu Leu Gly Ser Ala Val Leu Gly

50 55 60

Leu Leu Leu Val Leu Ala Ala Val Val Ala Trp Cys Tyr Tyr Ser Val

65 70 75 80

Ser Leu Arg Lys Ala Glu Arg Leu Arg Ala Glu Leu Leu Asp Leu Lys

85 90 95

Ala Gly Gly Phe Ser Ile Arg Asn Gln Lys Gly Glu Gln Val Phe Arg

100 105 110

Leu Ala Phe Arg Ser Gly Ala Leu Asp Leu Asp Ser Cys Ser Arg Asp

115 120 125

Gly Ala Leu Leu Gly Cys Ser Leu Thr Ala Asp Gly Leu Pro Leu His

130 135 140

Phe Phe Ile Gln Thr Val Arg Pro Lys Asp Thr Val Met Cys Tyr Arg

145 150 155 160

Val Arg Trp Glu Glu Ala Ala Pro Gly Arg Ala Val Glu His Ala Met

165 170 175

Phe Leu Gly Asp Ala Ala Ala His Trp Tyr Gly Gly Ala Glu Met Arg

180 185 190

Thr Gln His Trp Pro Ile Arg Leu Asp Gly Gln Gln Glu Pro Gln Pro

195 200 205

Phe Val Thr Ser Asp Val Tyr Ser Ser Asp Ala Ala Phe Gly Gly Ile

210 215 220

Leu Glu Arg Tyr Trp Leu Ser Ser Arg Ala Ala Ala Ile Lys Val Asn

225 230 235 240

Asp Ser Val Pro Phe His Leu Gly Trp Asn Ser Thr Glu Arg Ser Leu

245 250 255

Arg Leu Gln Ala Arg Tyr His Asp Thr Pro Tyr Lys Pro Pro Ala Gly

260 265 270

Arg Ala Ala Ala Pro Glu Leu Ser Tyr Arg Val Cys Val Gly Ser Asp

275 280 285

Val Thr Ser Ile His Lys Tyr Met Val Arg Arg Tyr Phe Asn Lys Pro

290 295 300

Ser Arg Val Pro Ala Pro Glu Ala Phe Arg Asp Pro Ile Trp Ser Thr

305 310 315 320

Trp Ala Leu Tyr Gly Arg Ala Val Asp Gln Asp Lys Val Leu Arg Phe

325 330 335

Ala Gln Gln Ile Arg Leu His His Phe Asn Ser Ser His Leu Glu Ile

340 345 350

Asp Asp Met Tyr Thr Pro Ala Tyr Gly Asp Phe Asp Glu Val Lys Phe

355 360 365

Pro Asn Ala Ser Asp Met Phe Arg Arg Leu Arg Asp Ala Gly Phe Arg

370 375 380

Val Thr Leu Trp Val His Pro Phe Val Asn Tyr Asn Ser Ser Arg Phe

385 390 395 400

Gly Glu Gly Val Glu Arg Glu Leu Phe Val Arg Glu Pro Thr Gly Arg

405 410 415

Leu Pro Ala Leu Val Arg Trp Trp Asn Gly Ile Gly Ala Val Leu Asp

420 425 430

Phe Thr His Pro Lys Ala Arg Asp Trp Phe Gln Gly His Leu Arg Arg

435 440 445

Leu Arg Ser Arg Tyr Ser Val Ala Ser Phe Lys Phe Asp Ala Gly Glu

450 455 460

Val Ser Tyr Leu Pro Arg Asp Phe Ser Thr Tyr Arg Pro Leu Pro Asp

465 470 475 480

Pro Ser Val Trp Ser Arg Arg Tyr Thr Glu Met Ala Leu Pro Phe Phe

485 490 495

Ser Leu Ala Glu Val Arg Val Gly Tyr Gln Ser Gln Asn Ile Ser Cys

500 505 510

Phe Phe Arg Leu Val Asp Arg Asp Ser Val Trp Gly Tyr Asp Leu Gly

515 520 525

Leu Arg Ser Leu Ile Pro Ala Val Leu Thr Val Ser Met Leu Gly Tyr

530 535 540

Pro Phe Ile Leu Pro Asp Met Val Gly Gly Asn Ala Val Pro Gln Arg

545 550 555 560

Thr Ala Gly Gly Asp Val Pro Glu Arg Glu Leu Tyr Ile Arg Trp Leu

565 570 575

Glu Val Ala Ala Phe Met Pro Ala Met Gln Phe Ser Ile Pro Pro Trp

580 585 590

Arg Tyr Asp Ala Glu Val Val Ala Ile Ala Gln Lys Phe Ala Ala Leu

595 600 605

Arg Ala Ser Leu Val Ala Pro Leu Leu Leu Glu Leu Ala Gly Glu Val

610 615 620

Thr Asp Thr Gly Asp Pro Ile Val Arg Pro Leu Trp Trp Ile Ala Pro

625 630 635 640

Gly Asp Glu Thr Ala His Arg Ile Asp Ser Gln Phe Leu Ile Gly Asp

645 650 655

Thr Leu Leu Val Ala Pro Val Leu Glu Pro Gly Lys Gln Glu Arg Asp

660 665 670

Val Tyr Leu Pro Ala Gly Lys Trp Arg Ser Tyr Lys Gly Glu Leu Phe

675 680 685

Asp Lys Thr Pro Val Leu Leu Thr Asp Tyr Pro Val Asp Leu Asp Glu

690 695 700

Ile Ala Tyr Phe Thr Trp Ala Ser

705 710

<210> 21

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 21

ctcaggttcc agtattcatc 20

<210> 22

<211> 21

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 22

gtagcacatg accgtgtcct t 21

<210> 23

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 23

tccgcaatca gaagggagag 20

<210> 24

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 24

cgccgatgcc gttccaccag 20

<210> 25

<211> 24

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 25

cagccacctg gaaatcgacg acat 24

<210> 26

<211> 24

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 26

gcaggagatg ttctgtgact ggta 24

<210> 27

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 27

cgcgggactt cagcacctac 20

<210> 28

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 28

aagtcagcag ccacttaatg 20

Claims (3)

1. An application of a MYORG mutant gene with c.103a > G mutation as a molecular marker in preparing a reagent for diagnosing primary familial cerebral calcification of human being, which is characterized in that: the coding sequence of the MYORG mutant gene is shown as SEQ ID NO:4, and the coding sequence of the MYORG wild type gene is shown as SEQ ID NO:1, then SEQ ID NO:4, mutating the gene contained in the MYORG mutant gene coding sequence shown in the formula 4 into c.103A > G; SEQ ID NO:4, the occurrence of a diploid homozygous form of the MYORG mutant gene shown in sequence 4 results in the occurrence of primary familial cerebral calcification in humans; SEQ ID NO:2-3 and any one of the MYORG mutant genes shown in SEQ ID NO:4, all the occurrence of diploid heterozygous forms composed of MYORG mutant genes shown in the sequence of the gene sequence can lead to the occurrence of primary familial cerebral calcification of human beings; SEQ ID NO:5-10 and any one of the MYORG mutant genes shown in SEQ ID NO:4, all appear in diploid heterozygous form composed of the MYORG mutant gene shown in the sequence of 4, which can lead to the occurrence of primary familial cerebral calcification in human.

2. The use according to claim 1, characterized in that the agent for diagnosing human primary familial cerebral calcification comprises the amino acid sequence as set forth in SEQ ID NO:21 and SEQ ID NO:22, capable of amplifying a primer set of the sequence set forth in SEQ ID NO:4, and a c.103A > G mutant fragment contained in the coding sequence of the MYORG mutant gene.

3. An application of a MYORG mutant gene coding protein with c.103A > G mutation as a molecular marker in preparing a reagent for diagnosing primary familial cerebral calcification of human beings, which is characterized in that: the detection target of the reagent for diagnosing human primary familial cerebral calcification is MYORG mutant gene encoding protein with c.103A > G mutation, which corresponds to SEQ ID NO:4, the coding sequence of the MYORG mutant gene is shown as SEQ ID NO:14, and the MYORG wild-type gene encodes a protein sequence as shown in SEQ ID NO:11, the corresponding amino acid is mutated to p.m35v; SEQ ID NO:11, and the sequence of SEQ ID NO: the occurrence of protein encoded by the MYORG mutant gene shown in the 14 sequence can reveal the occurrence of primary familial cerebral calcification.

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