KR100528627B1 - Genetic marker for decreased activity of cytochrome P450 3A7 protein - Google Patents

Abstract

The present invention relates to a gene as a marker associated with dysfunction of the CYP3A7 protein, which encodes a variant CYP3A7 protein encoding a translational termination codon (TAA) at the 55th amino acid position in the wild-type CYP3A7 amino acid sequence of SEQ ID NO: 21. Genes as markers associated with dysfunction of proteins are provided. By examining the retention of the gene, it is possible not only to predict individual differences in drug response of drugs metabolized by CYP3A7, but also to diagnose diseases related to CYP3A7 protein abnormalities.

Description

Gene marker for reduced activity of cytochrome P450 3A7 protein

The present invention relates to a gene as a marker associated with dysfunction of the CYP3A7 protein, and more particularly, to a CYP3A7 protein encoding a variant CYP3A7 protein in which a translation termination codon (TAA) is produced at the 55th amino acid position in a wild-type CYP3A7 amino acid sequence. The present invention relates to genes as markers related to protein dysfunction.

Cytochrome P450 3A7 protein (CYP3A7), along with CYP3A4, CYP3A5 and CYP3A43, is a drug metabolic enzyme belonging to the CYP3 family. CYP3A7 is produced mainly in the liver of the fetus and accounts for about 30% of the total amount of cytochrome P450 enzymes. This enzyme is known to be expressed in the embryonic stage of early pregnancy. CYP3A7 enzyme is involved in the metabolism of foreign body foreign material coming from outside of the fetus. In addition, it is known to be involved in the internal metabolism of testosterone 6-hydroxylation and dehydroepiandristerone 3-sulfate 16-hydroxylation (Hakkola J et al., Harmacol Toxicol. 1998, 82 (5): 209-217).

Single nucleotide polymorphism (SNPs), on the other hand, is a phenomenon in which the bases that make up the genome are partially different from person to person, and human SNPs are 0.1% of the human genome of about 3 billion base pairs, or 1 per 1,000 bases. It is estimated that about 3 million dogs exist (Cooper, DN et al., Hum. Genet. 1985, 69: 201-205). SNPs are the most common form of DNA sequence variation, especially those that occur in the nucleotide sequence of the region where the gene is encoded (the region that makes the protein from the gene), which in humans is associated with many diseases, including cancer, hypertension, diabetes, and dementia. It is reported to be closely related (Sherrington et al. , Nature 1995, 375: 754-760; Kunz et al ., Hypertension. 1997, 30: 1331-1337; Rogaev et al ., Genomics 1997, 40: 415- 424). Thus, when comparing SNPs between the normal population and the patient population, if specific SNPs appear to be associated with the disease, these SNPs can be used as very useful genetic markers for specific diseases (Lai, E., Genomics 1998, 54: 31-38). That is, by identifying one or more sequences specific to the genetic disease in DNA extracted from the cells of the individual, it is possible to diagnose the onset and possibility of the genetic disease of the individual.

The gene of CYP3A7 is known to exhibit a genetic polymorphism. To date, four mutations have been found in the promoter portion of the gene (see www.imm.ki.se/cypalleles/cyp3a7.htm). However, there is no known single-base polymorphism at the splicing site involved in the exon or mRNA process of protein encoding. CYP3A7 is expressed not only in the fetus but also in some adults and is known to play a role in the metabolism of external drugs. Gene mutations that alter the function of this enzyme can have undesirable consequences on the metabolism of drugs or biomolecules. (Hakkola J et al., Pharmacol Toxicol. 1998 May; 82 (5): 209-17). Specifically, the CYP3A7 gene mutation may show differences in pharmacokinetics for some drugs, may be sensitive to external toxicants, and may alter normal production by altering the production of biomaterials required during development. Therefore, CYP3A7 gene mutations must be identified, and among them, the identification of functional gene mutations that can change the function of the enzyme is more important. In addition, in the case of many genes, the distribution of genetic polymorphisms is known to be different according to the species. Therefore, it is necessary to confirm the frequency of the CYP3A7 gene even if there are no intrinsic gene mutations frequently occurring in Koreans.

Therefore, the present inventors conducted a study for discovering a single base polymorphism affecting the activity of CYP3A7 from Korean genes. As a result, another base was inserted into the nucleotide sequence encoding the wild type CYP3A7 protein. The present invention has been accomplished by finding a single base mutation in which a translation termination codon is generated at an amino acid position, and finding that the single base mutation is a Korean specific DNA marker associated with the enzymatic abnormality of the CYP3A7 variant protein.

It is therefore an object of the present invention to provide a gene as a marker associated with dysfunction of the CYP3A7 protein, which encodes a variant CYP3A7 protein which produces a translational stop codon (TAA) at the 55th amino acid position in the wild type CYP3A7 amino acid sequence.

In order to achieve the above object, the present invention is a labeling factor associated with the dysfunction of CYP3A7 protein, which encodes a variant CYP3A7 protein that generates a translation termination codon (TAA) at the 55th amino acid position in the amino acid sequence of SEQ ID NO: 21 Gene as.

Hereinafter, the present invention will be described in detail.

Genetic diseases develop due to changes in the nucleotide sequence of a normal gene. Such a change may be caused by substitution of one base of a gene with another base, or by insertion of another base between bases of the gene.

The present invention relates to a marker that can detect a genetic disease caused by insertion of another base between the bases of a gene. Specifically, the translation termination codon at the 55th amino acid position of the amino acid sequence of the wild type CYP3A7 protein of SEQ ID NO: 21. It is characterized by providing a single base mutation in which (TAA) is produced. Specifically, such a single base mutation may be caused by inserting a thymine (T) base between the 134th base (C) and the 135th base (T) in the nucleotide sequence of SEQ ID NO: 20 encoding the wild type CYP3A7 protein.

In one embodiment of the present invention, the entire nucleotide sequence of CYP3A7 genomic DNA (gDNA) represented by SEQ ID NO: 24 was analyzed using DNA isolated from the subject and an automatic sequencing analyzer (ABI 3700). A new single base polymorphism (SNP) was found.

Monobasic polymorphism according to the present invention is caused by the insertion of thymine (T) between the 22nd base (C) and 23rd base (T) of the second exon (7063 ~ 7155 bp) of CYP3A7 gDNA of SEQ ID NO: 24 When this corresponds to the CYP3A7 cDNA nucleotide sequence represented by SEQ ID NO: 20, it can be seen that the thymine (T) base is inserted between the 134th base (C) and the 135th base (T) (see FIG. 2). ).

When thymine is inserted in positions 134 and 135 of the CYP3A7 cDNA nucleotide sequence represented by SEQ ID NO: 20, the nucleotide sequence shown in SEQ ID NO: 22 will be obtained, and as shown in SEQ ID NO: 23, thereafter, A translational stop codon of TAA is generated at the 55th amino acid position that emerges. Therefore, the gene containing the mutation does not produce a complete form of the CYP3A7 enzyme but only produces a protein fragment that lacks the function. In a subject with the genetic mutation, the enzyme activity of the CYP3A7 is deficient, resulting in metabolic abnormalities of drugs or biological molecules. May result.

Therefore, by examining the possession of the CYP3A7 mutant gene, it is possible not only to predict individual drug response differences of drugs metabolized by CYP3A7, but also to diagnose diseases related to CYP3A7 protein abnormalities.

As a method for testing a gene as a marker associated with a dysfunction of the CYP3A7 protein according to the present invention, all conventionally known SNPs test methods can be used, and specifically, polymerase chain reaction-restriction fragment length formation (Polymerase Chain Reaction). Restriction Fragment Length Polymorphism (PCR-RFLP), Single Strand Conformation Polymorphism (SSCP), DNA sequencing, pyrosequencing, denaturation gradient gel electrophoresis gradient gel electrophoresis), DNA chips using oligonucleotides, and the like.

Specifically, pyro sequencing is a method of detecting which base is newly used according to each genotype during the DNA polymerization reaction.

In the case of bases used for DNA polymerization, pyrophosphate is advantageous, which is converted into ATP in the presence of adenosine 5'-phosphosulfate and luminescence reaction by luciferase activated by ATP. This is measured. Since the height of each peak is proportional to the number of bases added, the homozygote is twice the heterozygote peak. If the added base is not used for DNA synthesis, each genotype can be known because pyrophosphate is not advantageous and does not cause a luminescence reaction.

Specifically, the method for detecting a gene as a marker associated with dysfunction of the CYP2J2 protein of the present invention comprises the steps of: (1) separating DNA from a blood sample of a subject; (2) amplifying the separated DNA by performing PCR using primers of SEQ ID NO: 17 and SEQ ID NO: 18; And (3) identifying the mutation of the CYP3A7 gene by pyrosequencing using the primer of SEQ ID NO: 19 as a template of the PCR product of step (2).

In another embodiment of the present invention, the CYP3A7 gene mutation was searched for in the subject using the detection method. As a result, one of the two strands of DNA has a variant and one strand has a wild type. It could be confirmed (see Fig. 3).

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Amplification of the CYP3A7 Gene

After separating blood from 48 healthy subjects, DNA was isolated using Qiagen's Genomic DNA Separation Kit according to the instructions.

The entire nucleotide sequence of CYP3A7 gDNA represented by SEQ ID NO: 24 and the gDNA sequence isolated from the subject were analyzed using an automatic sequencing analyzer (ABI 3700) to find a new single base polymorphism (SNP). To determine whether this nucleotide polymorphism is located on the strand of the CYP3A7 gene, haplotype determination, whether there are no other gene mutations on the same DNA strand, or whether it originates from pseudogenes located on different parts of the chromosome. One strand of DNA from the subject's DNA containing the mutation was cloned in full size using a PCR reaction and then sequenced.

As listed in SEQ ID NO: 24, CYP3A7 gDNA has 13 exons and has very long introns between each exon, resulting in 33,226 bp of total gene length, resulting in four CYP3A7 gDNAs (3A7-P1, PCR was performed by dividing into fragments of P2, P3, P4). The primer pair used for each PCR, the binding position of the primer pair, and the size of the amplified product are shown in Table 1 below. In Table 1 below, “IVS” means an intron (intervening sequence), a number after IVS is a number indicating the number of introns, and also a “+ number” following an intron number (eg IVS2, IVS8, etc.). Indicates the relative position in the dowstream direction when the start of the intron is +1, and the “-number” following the intron number is upstream when the last position of the intron is -1. Relative position in the () direction.

 CYP3A7 gene fragment Primer pairs Position Size (bp) 3A7-P1 SEQ ID NO: 1 -665 to -646 4978 SEQ ID NO: 2 IVS2 + 156 to IVS2 + 175 3A7-P2 SEQ ID NO: 3 IVS2-135-IVS2-112 8217 SEQ ID NO: 4 IVS8 + 269-IVS8 + 288 3A7-P3 SEQ ID NO: 5 IVS8-254 ~ IVS8-235 4843 SEQ ID NO: 6 IVS11 + 63 to IVS11 + 82 3A7-P4 SEQ ID NO: 7 IVS10-231 to IVS10-211 5596 SEQ ID NO: 8 IVS13 + 1128 to IVS13 + 1147

On the other hand, PCR reaction conditions of each CYP3A7 gene fragment are shown in Table 2 below.

PCR products Reaction condition 3A7-P1 94 ° C 1 minute, (98 ° C 10 seconds, 58 ° C 30 seconds, 72 ° C 5 minutes) 30cycles, 72 ° C 10 minutes 3A7-P2 94 ° C 1 minute, (98 ° C 10 seconds, 58 ° C 30 seconds, 72 ° C 8 minutes) 30cycles, 72 ° C 10 minutes 3A7-P3 94 ° C 1 minute, (98 ° C 10 seconds, 60 ° C 30 seconds, 72 ° C 5 minutes) 30cycles, 72 ° C 10 minutes 3A7-P4 94 ° C 1 minute, (98 ° C 10 seconds, 56 ° C 30 seconds, 72 ° C 5 minutes) 30cycles, 72 ° C 10 minutes

The PCR product after the PCR reaction was confirmed by gel electrophoresis. As a result, as shown in Figure 1, it was confirmed that all of the PCR products of the 3A7-P1 to 3A7-P4 amplified.

Example 2 CYP3A7 Sequencing

Each PCR product obtained in Example 1 was confirmed by an automatic base sequence analyzer (ABI 3700). The sequencing primer sequence used at this time is shown in Table 3 below.

PCR products Sequencing primers 3A7-P1 SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 9 3A7-P2 SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 3A7-P3 SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 14 3A7-P4 SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 15

Autobase sequencing revealed no other mutations in the CYP3A7 gene DNA strand. However, as shown in FIG. 2, as in the first discovery, the mutant gene was confirmed that thymine (T) was inserted between the 22nd base and the 23rd base of the second exon. This corresponds to between 7084 and 7085 bases in the total CYP3A7 gDNA represented by SEQ ID NO: 24, and between 134 and 135 bases in the CYP3A7 cDNA represented by SEQ ID NO: 20.

On the other hand, a subject having such a mutant gene, as shown in Figure 3, one of the two strands of DNA was found to have a variant, one strand has a wild type.

When the thymine (T) base is inserted between the 134th base (C) and the 135th base (T) in the nucleotide sequence of SEQ ID NO: 20, the translation termination codon of TAA is generated at the 55th amino acid position subsequent thereto (sequence) Number 23). Thus, the gene containing the mutation does not produce a complete form of the CYP3A7 enzyme, but only produces a protein fragment lacking function. In subjects with such genetic mutations, the enzyme activity of CYP3A7 is deficient and may cause metabolic abnormalities of drugs or biological molecules.

Example 3 Detection of Mutants Using PCR / Pyro Sequencing

We searched for CYP3A7 gene mutations in blood samples of subjects. After preparing a primer pair for PCR so that a PCR product containing a second exon of CYP3A7 gDNA containing a mutation was produced, PCR was performed. As primer pairs, primers of SEQ ID NO: 17 and SEQ ID NO: 18 were used, and the 5 'side of SEQ ID NO: 17 was labeled with biotin. The PCR reaction was denatured at 94 ° C. for 5 minutes, and then the reaction was repeated 35 times for 30 seconds at 94 ° C., 30 seconds at 60 ° C., and 30 seconds at 72 ° C., and finally amplified at 72 ° C. for 5 minutes.

Then, it was confirmed that only one type of PCR product was generated by electrophoresis. The PCR product was used as a template to determine the presence or absence of mutations in a pyrosequencer (Pyrosequencing) using a sequencing primer of SEQ ID NO: 19. It was.

As a result, as shown in Figure 3, it can be seen that the two genotypes are distinct. In addition, pyro sequencing method was confirmed that the presence or absence of genetic mutation within 40 minutes starting from the PCR product.

Mutant genes identified through sequencing were used as controls to search for the presence of mutant genes in Koreans, Vietnamese, African Americans, and Westerners. The DNA of other races other than Koreans was provided by the Gene Bank, which is being built at the Inje University School of Medicine.

A total of 544 persons, 185 Koreans, 159 Vietnamese, 100 African-Americans and 100 Westerners, were tested for the presence of the mutant gene using a pyrosequencer. No subjects with the mutant gene were found. In conclusion, the frequency of this new gene mutation was 0.21% in Koreans.

As described above, the monobasic polymorphism found in the gene of cytochrome P450 3A7 (CYP3A7) first discovered in Korean can be applied to predict individual drug response differences of drugs metabolized by CYP3A7, and furthermore, CYP3A7 protein aberration-related diseases. It can be applied to diagnose.

1 is a gel photograph showing the PCR amplification product of the CYP3A7 gene.

Lane 1: Size Marker Treated with Lambda DNA with Hind III

Lanes 2 to 5: P1 to P4

Figure 2 is a result of analyzing the heterozygous variant gene of CYP3A7 using an automatic sequence analyzer, the arrow indicates the position where thymine is inserted.

3 is a wild-type and heterozygous mutation using the pyroquencer (Pyrosequencer) whether the thymine (T) base is inserted between the 134th base (C) and 135th base (T) in the base sequence of SEQ ID NO: 20 The gene was confirmed.

<110> INJE UNIVERSITY <120> Genetic marker for decreased activity of cytochrome P450 3A7 protein <130> 2004006 <160> 24 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P1 <400> 1 aaacaggcct ggaaacacac 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P1 <400> 2 gagttagcaa gagagcccca 20 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P2 <400> 3 gacatctcta aataagcttc ctct 24 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P2 <400> 4 agggaagtgc tgtgataagg 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P3 <400> 5 gggtatgcag ttaggaccag 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P3 <400> 6 gcccagactg tcctgtagag 20 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P4 <400> 7 gttagagcct tcccgaatgt c 21 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer for CYP3A7-P4 <400> 8 ttttagtaga aagggggctt 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 9 tgtgataaga acccagaacc 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 10 atgtaatgtc cggatcaaag 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 11 tgatgcctgt ttaacacatt 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 12 gttcctttcc gaggagtagt 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 13 gacatgtgga tttcctgctg 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 14 catctaaact gtgatgccct 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 15 gtgggcccta agtaagaaac 20 <210> 16 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 16 cgaatccaag atttatagtg c 21 <210> 17 <211> 23 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 17 gatcattcca gttcctcagt cag 23 <210> 18 <211> 24 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 18 gcaatcataa gaagcaaaag agga 24 <210> 19 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> sequencing primer <400> 19 cctctgcctt tttt 14 <210> 20 <211> 1512 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (1509) <400> 20 atg gat ctc atc cca aac ttg gcc gtg gaa acc tgg ctt ctc ctg gct 48 Met Asp Leu Ile Pro Asn Leu Ala Val Glu Thr Trp Leu Leu Leu Ala 1 5 10 15 gtc agc ctg ata ctc ctc tat cta tat gga acc cgt aca cat gga ctt 96 Val Ser Leu Ile Leu Leu Tyr Leu Tyr Gly Thr Arg Thr His Gly Leu 20 25 30 ttt aag aag ctt gga att cca ggg ccc aca cct ctg cct ttt ttg gga 144 Phe Lys Lys Leu Gly Ile Pro Gly Pro Thr Pro Leu Pro Phe Leu Gly 35 40 45 aat gct ttg tcc ttc cgt aag ggc tat tgg acg ttt gac atg gaa tgt 192 Asn Ala Leu Ser Phe Arg Lys Gly Tyr Trp Thr Phe Asp Met Glu Cys 50 55 60 tat aaa aag tat aga aaa gtc tgg ggt att tat gac tgt caa cag cct 240 Tyr Lys Lys Tyr Arg Lys Val Trp Gly Ile Tyr Asp Cys Gln Gln Pro 65 70 75 80 atg ctg gct atc aca gat ccc gac atg atc aaa aca gtg cta gtg aaa 288 Met Leu Ala Ile Thr Asp Pro Asp Met Ile Lys Thr Val Leu Val Lys 85 90 95 gaa tgt tat tct gtc ttc aca aac cgg agg cct ttc ggg cca gtg gga 336 Glu Cys Tyr Ser Val Phe Thr Asn Arg Arg Pro Phe Gly Pro Val Gly 100 105 110 ttt atg aaa aat gcc atc tct ata gct gag gat gaa gaa tgg aag aga 384 Phe Met Lys Asn Ala Ile Ser Ile Ala Glu Asp Glu Glu Trp Lys Arg 115 120 125 ata cga tca ttg ctg tct cca aca ttc acc agc gga aaa ctc aag gag 432 Ile Arg Ser Leu Leu Ser Pro Thr Phe Thr Ser Gly Lys Leu Lys Glu 130 135 140 atg gtc cct atc att gcc cag tat gga gat gtg ttg gtg aga aat ctg 480 Met Val Pro Ile Ile Ala Gln Tyr Gly Asp Val Leu Val Arg Asn Leu 145 150 155 160 agg cgg gaa gca gag aca ggc aag cct gtc acc ttg aaa cac gtc ttt 528 Arg Arg Glu Ala Glu Thr Gly Lys Pro Val Thr Leu Lys His Val Phe 165 170 175 ggg gcc tac agc atg gat gtg atc act agc aca tca ttt gga gtg agc 576 Gly Ala Tyr Ser Met Asp Val Ile Thr Ser Thr Ser Phe Gly Val Ser 180 185 190 atc gac tct ctc aac aat cca caa gac ccc ttt gtg gaa aac acc aag 624 Ile Asp Ser Leu Asn Asn Pro Gln Asp Pro Phe Val Glu Asn Thr Lys 195 200 205 aag ctt tta aga ttt aat cca tta gat cca ttc gtt ctc tca ata aaa 672 Lys Leu Leu Arg Phe Asn Pro Leu Asp Pro Phe Val Leu Ser Ile Lys 210 215 220 gtc ttt cca ttc ctt acc cca att ctt gaa gca tta aat atc act gtg 720 Val Phe Pro Phe Leu Thr Pro Ile Leu Glu Ala Leu Asn Ile Thr Val 225 230 235 240 ttt cca aga aaa gtt ata agt ttt cta aca aaa tct gta aaa cag ata 768 Phe Pro Arg Lys Val Ile Ser Phe Leu Thr Lys Ser Val Lys Gln Ile 245 250 255 aaa gaa ggt cgc ctc aaa gag aca caa aag cac cga gtg gat ttc ctt 816 Lys Glu Gly Arg Leu Lys Glu Thr Gln Lys His Arg Val Asp Phe Leu 260 265 270 cag ctg atg att gac tct cag aat tca aaa gac tct gag acc cac aaa 864 Gln Leu Met Ile Asp Ser Gln Asn Ser Lys Asp Ser Glu Thr His Lys 275 280 285 gct ctg tct gat ctg gag ctc atg gcc caa tca att atc ttt att ttt 912 Ala Leu Ser Asp Leu Glu Leu Met Ala Gln Ser Ile Ile Phe Ile Phe 290 295 300 gct ggc tat gaa acc acg agc agt gtt ctc tcc ttc att ata tat gaa 960 Ala Gly Tyr Glu Thr Thr Ser Ser Val Leu Ser Phe Ile Ile Tyr Glu 305 310 315 320 ctg gcc act cac cct gat gtc cag cag aaa gtg cag aag gaa att gat 1008 Leu Ala Thr His Pro Asp Val Gln Gln Lys Val Gln Lys Glu Ile Asp 325 330 335 aca gtt tta ccc aat aag gca cca ccc acc tat gat act gtg cta cag 1056 Thr Val Leu Pro Asn Lys Ala Pro Pro Thr Tyr Asp Thr Val Leu Gln 340 345 350 ttg gag tat ctt gac atg gtg gtg aat gaa aca ctc aga tta ttc cca 1104 Leu Glu Tyr Leu Asp Met Val Val Asn Glu Thr Leu Arg Leu Phe Pro 355 360 365 gtt gct atg aga ctt gag agg gtc tgc aaa aaa gat gtt gaa atc aat 1152 Val Ala Met Arg Leu Glu Arg Val Cys Lys Lys Asp Val Glu Ile Asn 370 375 380 ggg atg ttt att ccc aaa ggg gtg gtg gtg atg att cca agc tat gtt 1200 Gly Met Phe Ile Pro Lys Gly Val Val Val Met Ile Pro Ser Tyr Val 385 390 395 400 ctt cat cat gac cca aag tac tgg aca gag cct gag aag ttc ctc cct 1248 Leu His His Asp Pro Lys Tyr Trp Thr Glu Pro Glu Lys Phe Leu Pro 405 410 415 gaa agg ttc agt aaa aag aac aag gac aac ata gat cct tac ata tac 1296 Glu Arg Phe Ser Lys Lys Asn Lys Asp Asn Ile Asp Pro Tyr Ile Tyr 420 425 430 aca ccc ttt gga agt gga ccc aga aac tgc att ggc atg agg ttt gct 1344 Thr Pro Phe Gly Ser Gly Pro Arg Asn Cys Ile Gly Met Arg Phe Ala 435 440 445 ctc gtg aac atg aaa ctt gct cta gtc aga gtc ctt cag aac ttc tcc 1392 Leu Val Asn Met Lys Leu Ala Leu Val Arg Val Leu Gln Asn Phe Ser 450 455 460 ttc aaa cct tgt aaa gaa aca cag atc ccc ctg aaa tta cgc ttt gga 1440 Phe Lys Pro Cys Lys Glu Thr Gln Ile Pro Leu Lys Leu Arg Phe Gly 465 470 475 480 gga ctt ctt cta aca gaa aaa ccc att gtt cta aag gct gag tca agg 1488 Gly Leu Leu Leu Thr Glu Lys Pro Ile Val Leu Lys Ala Glu Ser Arg 485 490 495 gat gag acc gta agt gga gcc t ga 1512 Asp Glu Thr Val Ser Gly Ala 500 <210> 21 <211> 503 <212> PRT <213> Homo sapiens <400> 21 Met Asp Leu Ile Pro Asn Leu Ala Val Glu Thr Trp Leu Leu Leu Ala 1 5 10 15 Val Ser Leu Ile Leu Leu Tyr Leu Tyr Gly Thr Arg Thr His Gly Leu 20 25 30 Phe Lys Lys Leu Gly Ile Pro Gly Pro Thr Pro Leu Pro Phe Leu Gly 35 40 45 Asn Ala Leu Ser Phe Arg Lys Gly Tyr Trp Thr Phe Asp Met Glu Cys 50 55 60 Tyr Lys Lys Tyr Arg Lys Val Trp Gly Ile Tyr Asp Cys Gln Gln Pro 65 70 75 80 Met Leu Ala Ile Thr Asp Pro Asp Met Ile Lys Thr Val Leu Val Lys 85 90 95 Glu Cys Tyr Ser Val Phe Thr Asn Arg Arg Pro Phe Gly Pro Val Gly 100 105 110 Phe Met Lys Asn Ala Ile Ser Ile Ala Glu Asp Glu Glu Trp Lys Arg 115 120 125 Ile Arg Ser Leu Leu Ser Pro Thr Phe Thr Ser Gly Lys Leu Lys Glu 130 135 140 Met Val Pro Ile Ile Ala Gln Tyr Gly Asp Val Leu Val Arg Asn Leu 145 150 155 160 Arg Arg Glu Ala Glu Thr Gly Lys Pro Val Thr Leu Lys His Val Phe 165 170 175 Gly Ala Tyr Ser Met Asp Val Ile Thr Ser Thr Ser Phe Gly Val Ser 180 185 190 Ile Asp Ser Leu Asn Asn Pro Gln Asp Pro Phe Val Glu Asn Thr Lys 195 200 205 Lys Leu Leu Arg Phe Asn Pro Leu Asp Pro Phe Val Leu Ser Ile Lys 210 215 220 Val Phe Pro Phe Leu Thr Pro Ile Leu Glu Ala Leu Asn Ile Thr Val 225 230 235 240 Phe Pro Arg Lys Val Ile Ser Phe Leu Thr Lys Ser Val Lys Gln Ile 245 250 255 Lys Glu Gly Arg Leu Lys Glu Thr Gln Lys His Arg Val Asp Phe Leu 260 265 270 Gln Leu Met Ile Asp Ser Gln Asn Ser Lys Asp Ser Glu Thr His Lys 275 280 285 Ala Leu Ser Asp Leu Glu Leu Met Ala Gln Ser Ile Ile Phe Ile Phe 290 295 300 Ala Gly Tyr Glu Thr Thr Ser Ser Val Leu Ser Phe Ile Ile Tyr Glu 305 310 315 320 Leu Ala Thr His Pro Asp Val Gln Gln Lys Val Gln Lys Glu Ile Asp 325 330 335 Thr Val Leu Pro Asn Lys Ala Pro Pro Thr Tyr Asp Thr Val Leu Gln 340 345 350 Leu Glu Tyr Leu Asp Met Val Val Asn Glu Thr Leu Arg Leu Phe Pro 355 360 365 Val Ala Met Arg Leu Glu Arg Val Cys Lys Lys Asp Val Glu Ile Asn 370 375 380 Gly Met Phe Ile Pro Lys Gly Val Val Val Met Ile Pro Ser Tyr Val 385 390 395 400 Leu His His Asp Pro Lys Tyr Trp Thr Glu Pro Glu Lys Phe Leu Pro 405 410 415 Glu Arg Phe Ser Lys Lys Asn Lys Asp Asn Ile Asp Pro Tyr Ile Tyr 420 425 430 Thr Pro Phe Gly Ser Gly Pro Arg Asn Cys Ile Gly Met Arg Phe Ala 435 440 445 Leu Val Asn Met Lys Leu Ala Leu Val Arg Val Leu Gln Asn Phe Ser 450 455 460 Phe Lys Pro Cys Lys Glu Thr Gln Ile Pro Leu Lys Leu Arg Phe Gly 465 470 475 480 Gly Leu Leu Leu Thr Glu Lys Pro Ile Val Leu Lys Ala Glu Ser Arg 485 490 495 Asp Glu Thr Val Ser Gly Ala 500 <210> 22 <211> 165 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (162) <400> 22 atg gat ctc atc cca aac ttg gcc gtg gaa acc tgg ctt ctc ctg gct 48 Met Asp Leu Ile Pro Asn Leu Ala Val Glu Thr Trp Leu Leu Leu Ala 1 5 10 15 gtc agc ctg ata ctc ctc tat cta tat gga acc cgt aca cat gga ctt 96 Val Ser Leu Ile Leu Leu Tyr Leu Tyr Gly Thr Arg Thr His Gly Leu 20 25 30 ttt aag aag ctt gga att cca ggg ccc aca cct ctg cct ttt ttt ggg 144 Phe Lys Lys Leu Gly Ile Pro Gly Pro Thr Pro Leu Pro Phe Phe Gly 35 40 45 aaa tgc ttt gtc ctt ccg taa 165 Lys Cys Phe Val Leu Pro 50 <210> 23 <211> 54 <212> PRT <213> Homo sapiens <400> 23 Met Asp Leu Ile Pro Asn Leu Ala Val Glu Thr Trp Leu Leu Leu Ala 1 5 10 15 Val Ser Leu Ile Leu Leu Tyr Leu Tyr Gly Thr Arg Thr His Gly Leu 20 25 30 Phe Lys Lys Leu Gly Ile Pro Gly Pro Thr Pro Leu Pro Phe Phe Gly 35 40 45 Lys Cys Phe Val Leu Pro 50 <210> 24 <211> 33226 <212> DNA <213> Homo sapiens <220> <221> exon (222) (3016) .. (3192) <220> <221> exon (222) (7063) .. (7155) <220> <221> exon <222> (15867) ... (15920) <220> <221> exon (222) (17804) .. (17902) <220> <221> exon (222) (20576) .. (20689) <220> <221> exon (222) (20950) .. (21039) <220> <221> exon <222> (22310) .. (22456) <220> <221> exon <222> (23533) .. (23660) <220> <221> exon <222> (24680) .. (24745) <220> <221> exon (222) (27322) .. (27483) <220> <221> exon <222> (28954) .. (29181) <220> <221> exon (222) (30242) .. (30403) <220> <221> exon <222> (32620) .. (33178) <400> 24 agaaagaaga aacgtctttc attgttgata aggacatgat ttttctcaca attttatgat 60 tatttttcct tagctgtcct ataattatct gcttatttgt ctcttctcca tgtgcttagg 120 gtacaaagtt gaccaagacc aagaataatg tctgggagca caatactgac agcacagctt 180 taaaaacatg atgaatgctt taatacagga aatgagtagg ggagaggcaa gtgttgcttg 240 ggtgttcttc caatgcatag tatcttcctt gacacagtca gtgcagctct cagtaggcaa 300 gtccctacat gttagaagat gttactttct gtggaattaa gtggcagaac atgccttcaa 360 ttattttcct ttgcagaaca acaccaattt cattagttag gacagagtgc tggctgcatt 420 tgaattccaa gcaacgatta gtctatcact gttggtatag attccaacca gtcacaccac 480 ctcctgaagt ttgtagggca ggtaaatctt catcttagaa taaaaatcat cttagccaag 540 taagtgtttt agaggaaaga agaaaacata atcgtttcca taagagtttt gtttctaaaa 600 aaataagaaa ggctctttgt ttaggtgagc taatgaagtt gttgatagtt atcagatgac 660 actggaatct ttacttgtca gaatgtgttc tgtgcacctc tcggtgtggc aacatagaga 720 gggagatcct ccagcaatgc cattgatatg gtcagaaact gcatctttct ttctccctgc 780 tgagatggag tcctttgttc tagaaaaccc agggggtgcc actgggagta acccttgaga 840 caggaacacg aatctcaacc aatttctggt tgcagccttg agtcttacta tttgccatag 900 tgatgcttag caaggaatgg caggtgcacc agagcagcag aggacctaat atctcccttc 960 ctgttaactt tttataatat tttattgtga tcagtatcag ttgggaagct acttgcagtc 1020 actgagcctc agtttctaca tctgtaaact ggggatagta gcatggccct atttaatgtg 1080 ctcagcgaag ccactgaaag gagacagaaa tgtaccagaa ttccctggac ttttatccta 1140 cttctcctgg ggattgtcac ccacctaccc gtgtctgtcc tttgttgctt tgacgctgtc 1200 acttcttttc ttaggtacct ctctgtaggg ctccattatt ccagggattc cagagttaca 1260 gcacatgcat acctccatcc aagcatgttt atttgtctcc tgcttcacta ggctgtcccc 1320 aaggaacatg tggctcccgg cacatacctg gcacaacact gcacatgaca ttcacccact 1380 tggccttgaa tctgacaagg aatctggcat gatgttcacc tgctgaggcc aggtgccgag 1440 cagccctgga ggcttagggg ccagagggat gggaaaaggt gtctttctgg ggtgagtatc 1500 agtttctgca ggagtgctga acctgagaaa gaataaagag agaaggaagt gaacaagcac 1560 agcttaaaca tcatctgttt ctactgagtt ttaacaactc tgagattttg tttgtcatgg 1620 aatccatttc tcaggccaag cagacacaga acttgggtgt gagtgatgat aatgagctga 1680 tataattttc acaccctcat cactgagatc tctcccatca ggaatgggtc acagggctca 1740 caggtggcag caactgttat tacaggcctc atctctacca gctcctggca cctgctctcc 1800 tcccattaga aaatcctcca cttgtcaaaa aggaagccat ttgctttgaa ttccaattcc 1860 accctcaaga ggctgggacc acctcattgg agtccttgat gctgtgtgac ctgcagtgac 1920 cactgcccca ttgttgctgg ctgaggtggt ttgggtcaac ctggccatct gggcagctgt 1980 tctcttctct tctttctccc ctactgtttc cagacatgca gtatttccag agagaagggg 2040 ccactctttg gcaaagaacc tgtctaactt tctatctacg gcaggacttt tgaaagctac 2100 agaggaagaa gcacaaattg atgctattcc actaagccat cagctccatc tcatccatgc 2160 catgtctctt ttttaggggt cctcttgcca acagaatcac agaggacaaa tctgaaagtg 2220 cagagacagc agctgaggca cagccaagag ctctggctgt attaatgacc taagaagatg 2280 gagtggtcac cagaaagtca gaggaagtga cacacagggg cccagcaatc tcagccaagt 2340 caactccacc agcctttctg gtccccactg tgtgtacagc accctgatag ggaccagagc 2400 catgagagtg agtaagacca gactatgccc ttgaggagct cacctctgct aagggaaaca 2460 ggcctggaaa cacacaatgg tggtaaagag gaaagaagac aatagaactg catgaagggg 2520 atggaaagtg cccaggggag gaaatggtta cttctgtgtg agggggttgg tgaggaaaga 2580 ctctaagaga aggctctgtc tggctgggta tgaaaggatg tgtaggagtc ttctaggggg 2640 cacaggcaca ctccaggcat aggtaaagat ctgtaggcat ggcttgttgg gatgagtttc 2700 aagtattctg gaatgaggac agccatagag acaagaggag agttaataga ttttatgcca 2760 atggctccac ttgagtttgt gataagaacc cagaaccctt ggactcccca gtaacattga 2820 ttgagttgtg tatgattcta catagaatat taactcaatg gaggtcagtg agtggtgtgt 2880 gtgtgattat ttgccaactg ccgaggtgga gaagcctctt ccgactgcag gcagagcacg 2940 ggggccctgc tactggctgc agctccagcc ctgcctcctt ctccagcata taaacaatcc 3000 aacagcctca ctgaatcact gctgtgcagg gcaggaaagc tccacacaca cagcccagca 3060 aacagcagca cgctgctgaa aaaaagactc agaggagaga gataaggaag gaaagtagtg 3120 atggatctca tcccaaactt ggccgtggaa acctggcttc tcctggctgt cagcctgata 3180 ctcctctatc tgtgagtaac tgttcaggct cctcttctct gtttccttgg acttggggtg 3240 ctaatcaggc ctctctttcc cttatctgtt ttgaatagca aaaaggatgt tcaggctggc 3300 cgtggtggct cacacctgtc atcccagaac tttgggatgc tgagcaggtg gactgcctga 3360 ggtcaggagt tcaagaccag cctggctaat atggtgaaac cccatctcta ctaaaattac 3420 aaaaattagc tgggcatggt agtgcataac tgtagtccca gctacttggg tggctgaggc 3480 aggagaattg cttgaacctg ggaggcggag gttgcagtga actgagatca tgccactgca 3540 ctccagcctg ggtaacagag tgagattcca tctcaaaaaa aaaaaaagag aaaaaaaaga 3600 aaaaaaaaga ttttcaagga gaagtagctt aagtgttgga tgctacaaac atatagaggt 3660 tatgatggat gttttgcaga tctataaagg aataaataag catctccccc attcatcttt 3720 agtggaaaga agggtttagg gacagcattg attgaggata atctaataga caatggtttg 3780 tacccagctt accaaaccag aatttggatt ttacattttt ccctttaagt tgcaggagaa 3840 gagggtgaat cttctcacca gtgagatcct ggcatcctag agcaggtgga gagaagagtc 3900 actccctact gtgtgtactg gaggcttccc gcctgtcaca ttttacttct aactcaattt 3960 cacagttatt aagagtaagg aatcatggtc acatggaaac agaaaataca aacctcgttt 4020 taattctttc acagaaaagt tagcaattca gtgagttgtg gtaacatagt ttccatctac 4080 tggcaattta acatgaattg gtgtggctta aattcatcct ttttaaagcc agattttttt 4140 ggagatagac tatttccagc atgttccttc tgagtgataa aagagggcta ttagttcagt 4200 atttgtgaca ataaatgtgt ataagataac ctcacctttc cagaataatg tcaggaatat 4260 gagtctaatg tacaaatata tatctctagg acaagactgc atatgtcttt ttaaaaatac 4320 tttcccaaga gtttatttta atacccccat ttcaaataca cctgcttagc aggttatcct 4380 aaactctcag ggctagaaag taagcaagac tgtgagccag tgatgatagc aaaagcatcc 4440 aggtaggatc aagatgaagt gagaaaatat tccttacccc tcagggtaga actccaaata 4500 gatattcatg ggtcctggcc ccctagtgga ggtcacttga aggacaaaca tgttggcatc 4560 tcatctgctt gaagcctgga cacaagggca ccatcagtgt cactctgtgt gtggctggcc 4620 atgttgtggg gctgtcacta cagattctgg cagtcaggca gacaatacct tagccttaga 4680 cgatgccggt acagcccagg attcagaagc tatagtgtag accatgccct tcttaggcct 4740 atacaattac atgcaataga tgatggattt ttctgttagc ctcttcattg taacaaaaag 4800 cagcattact ctaccaaaca gaggggagct ggaaagaaac taaacagttt gccaagcaca 4860 gcctctgcct tgacgtggaa tcatgtgaat ctagacactc acctagatct ttcctcgggg 4920 gccaatgctg ctcacacatt aactcaatag tttgtcctgg cctgagagat catgtaacat 4980 gtagaaagtt tagaagcaga gattagtgcc attgattttc catggctgtg acaacaaagg 5040 aaggaacagg agtgggaaaa cccaaggcca ccctggttgt ggtagatggt gcacacactt 5100 ccactaacta ttctggggca aggatccaaa tgcactattg ggcctggcta tgctgtctct 5160 gctgggttct ccaaacacga gcctccatgc tgtttcttag ttgtatttta ccacatatta 5220 tcagagtcac tggatttgta cagaatgttt ggaacctata ctgccttaat ggctaccctt 5280 taaagaagag aaaacaaggt cttaattcaa ctatctggaa cattttatgg ttacttatgt 5340 ggaatactcc atcttttgct atcaacagga aggtacctgg ccattagaag gtgcctgcct 5400 ttcagctgaa agcacatatg aggcatatgg atccatttat acacaccgta cttttgaacc 5460 acattttcct aatttgcctg tctggggcca agcttgtggg actagcagat tcatggttga 5520 attgatggct ggctacctct tccatcaact tttctgcttc atccagtctt ccacccctca 5580 gtaacctcaa actgggaagg tcttcagata tccagaatcc ccagttgggg aagtccatac 5640 atggcccatc aaagatgggt tgcaagcagg cctgctatag ggagcagcag ccttgatggg 5700 ttttcctaca gagacagttg atgtgcagac gcaataaact gactgctttg taattgatca 5760 ccttgaaaaa aaatgagcat gtctggctgt attagtcttt tcttccattg ctataaaaaa 5820 aaatctcagg gtgggtaatt tagaaaggaa agagatttaa ttggctcatg gttctgcagg 5880 ctctacagga agcatagcga cttctgccgc tagggtgccc tcaggaaatt tacaatcatg 5940 gcagaaggtg aaaaagaagc aggcagatct tacatgactg gagcagaagc aagagagggt 6000 ggggaggagg tgccacacac ttataaacca ccagatctca taaaaactca ttgtcaagag 6060 gacagcaccc aagggggatg gtgtgaaacc atgagaaact gcctctaaga tacaaccacc 6120 atctatcagg ccccacttcc agaattgggg attacatttc aacatgagat ttgggtaggg 6180 gcacagattc agatcatatc actggcactg tgcttatcag gtggatatca ccagttggaa 6240 ggctagattc cacaagagga ggaatgtcct tgttcttagt tgtgattctt ctgcacagtg 6300 tcattcagga aaatatgagt caatcatcct ccccaatagc tcaaatcaat cagatcctct 6360 gaccacagag actgaggtgt agctaaaagg tgctcgcact tctacgaggc caatggaagc 6420 ctggagcaca gttgtcaatc tgtagaaata aggactctgt gactcctcca agacctctct 6480 gtgaatgaca gtttaagaag ggccagatcc taaaacaggg tcagagctta gagtgaaggg 6540 aaagcttaaa agcctctgag caaattctaa agacagggtc accataggct ctcagtgacc 6600 ccctgtgact gagtggatgc agtgatgcaa aatctcatca tcactgcaga agacaaaaga 6660 aaaattgtca ctctttctac ctaggatgag aatcccaaaa tcagggaaga gtccacttac 6720 taaaaagaca tgagggaatg aagtgcctgg aagaattcgt tcctgaacct ctcaggatca 6780 ttcacatttg agaacattta tcaaatattc atcccagact ggggccatga agacttcagc 6840 tgcttttagc taatcattgt aactttttgg tgtctcatgg tggaagcatg aaaggacctg 6900 atgaacaaat ataatcattg ccgtcagagt tactgttatt ttttattgcc ttaatgttac 6960 ctcctttgcc tgatcattcc agttcctcag tcagtgactc accagcccct atatctataa 7020 agtcacaatc cctgagacct gatttttgtt tcactttgta gatatggaac ccgtacacat 7080 ggacttttta agaagcttgg aattccaggg cccacacctc tgcctttttt gggaaatgct 7140 ttgtccttcc gtaaggtgag tgtttttgag cttcctcttt tgcttcttat gattgcaaag 7200 agcaacttag ttccatcagt aaaaatgctt ctcctcaaga ggaaggtctg agattttata 7260 ctttcagaaa tagcgtgtag gcatcaccca gagcgtggca aagtttaccc tggggctctc 7320 ttgctaactc tcaggaacct caagtttgcc tcagttggac agcccaaatg tcaagtagat 7380 ccagcaatct gatgttcaca acttgatgtg cagactttgt agagcaccat aaagagcatt 7440 tcctttttgc tccaggcagg ttcccaggaa ggtacagcca tacttagtta gtattaaaag 7500 taggaacagg agcccctgat tgtggcttgt tatctgtgga catgagagta aaactttcct 7560 ctctgtagca agagctcatg aataggcttt tttcccttcc tcctgcacag cttggaaact 7620 cagtttatta gggagatcaa gtatttacca ttgggtcatt tgcacctgct tgaagcctct 7680 tcaaaaaatg ccaaattttg ttggggaatt tccatggctt taacccaggt cttactcccc 7740 atgacatctg ttgccacttt gagacacaga acaaaagtgt tctgctttac ttcttttcct 7800 ttgaaggctg cgtcttgtgg gtgagtctga gcagggagtg gatgtaactg agatgagata 7860 gtggttgtct attaacctca gatatttaag cacaatatga attcatgggt ttatttttta 7920 cacaaaggca gaatcacatg caaaatcact agaaagggga tgatttgaca aaattattga 7980 atatttaatt tcactaaata gaatgtcata tgtgcctcat ggactgggga ctttggtcga 8040 gcctttgtgc ctaaccttat ctgatgctgt gttcagggcc tggtccctgg atggaaaatt 8100 atgagcttat ttccacaggg ccagcagcag caggagtcag ctctctggct tagacaatct 8160 gacctgcata cctgaggctt gtttctcaga ttctagttct cctaagtgag ggctttgttg 8220 tctaattact gcccaggaat ttcatatttt ttccctctgc aaaagcaata atttccctac 8280 tttcacatgc gtccgtgtga ggagaccacc aaacaggctt tgtgtgagca ataaatgttt 8340 ttaatcactt gggtgcaggc caactgagtc tgaaaaagga gtcagcaagg ggagataggg 8400 gtggggcagt tttataggag tggggtaagc agtggaaagt tacagttaaa ggtggttatc 8460 cattgtcagc agtggagggg gccacaaggt gcatggtggg gagatcataa gactcattgt 8520 gcagagaaga atgtcaccag gttgattgat caatcagttg ggacagggca ggaacaagtc 8580 ataatggaat gtcgtaagat tgctcaatca gttaagacag gagctggctg tttcatttct 8640 tttgcagttt tcagttgcct caggccatct ggatgtacac gtgcaggctt gggctcagag 8700 gcctgacatt cctgtctttt tatattaaga aaaataaaac aaaatagtgg caaagtgttg 8760 gggcagcaaa aatttttggg gggtggtatg gagagataat ggatgatgtt tctcagggct 8820 gcttcaagcg ggattatggg ccatgtggac accttaaaga aaattttata atgagttaca 8880 aggaatagga atttaggctg tggggagatc ttggggcaga ggatggtacc atggacttgt 8940 taaaagtaat atttgttgta tagaataatt ggtgatagtc tgaatgtgtt tttatattaa 9000 ttgagaaacc aattgaagac acaagtcccg aataacagaa ggagaaaaac aggtaccaga 9060 ggactaagaa tagggaggag ctaagacacc cagttagaaa gtgtccaggt gggtccaaaa 9120 taagtatttg cctggctggt gagtttttgg tctttatctt taagcttttt tatattgtca 9180 tataccaggc cagattggtt tagataaaaa caacattctt catttaaaaa tatagagtac 9240 tcctttttca gcagtgagta ggtcaaggcc tcagcgatgt tggaggacga ctagggctaa 9300 gaagtcaacc tgggtctgaa ggactgataa agtttgtgat gtatctgtaa tgctagcaga 9360 aaagtcatta gaaaggctac agaaagttgt gacagaagtt gaaatacctg ctattccagc 9420 tccaagggca atagtggaag cagaaagtcc taaccctaca agtagaggga ttagaggaat 9480 aactcatttt tgtctggttg gtgtcatgag ggggacagga agttgttcag tcccatttgc 9540 aaattgaatt ttttcaattt gggaataaga aatactagtg tacatgtacc tgtccaatta 9600 gcagatagac atatgtacgt ggtggagcca cagaggaagc agaggccttg tgtgaggcaa 9660 aactggaaat gcaaagtgaa aagatgagag ggagtgccaa aagaggtgtc ttgcacccag 9720 actcttaggg atttagtgag ggcatcagcc cttagtggtc ataatgggga ttgatggggc 9780 aactgggtat gagaaagagc atagtgtcta caaccaaact ttcattgcta ttcatggggt 9840 tcggtataag taaacaagaa gggggaatgg gaggagagtc tgaagaacaa ggggaaggta 9900 gccaagaatg gagtgaaata cagggcaaat gtcttaaggg aaatgagaag ttctaggagg 9960 gggctagttg cttgtaacct acatggaaga ggtcatgaaa tgacaacaga atagaatggg 10020 cctgtgagac tggaaggaga tatttttctt ggtctaagaa ccatcttcct tgagtgggga 10080 gggattgata gatggaaact tcagtgggag agtaaatagg agtgactgat gagaaggaga 10140 aaaactggcc atgagggaca gaagtaggaa tactggttgc ttctttagct gtcttatcag 10200 cataattgtt gccttgagca atggggtctg aggccctttg atggcctttg cagtgaatga 10260 ctccagcttt ttttggaagt aaagtggcct tgagaagaat ttttattaaa gaggcattaa 10320 tgatggagga cacttgcata gtgaagaaac ttctttcagc ccatattaca gcatggtggt 10380 gcaggatatg gacagcattt agagtcagta taaatattga tgtgtagtcc ctttgcaaga 10440 gtgaggcccc aagttaaggc aatgagtttg gcttgctgag agatagtgga ggggggcaga 10500 gcagtagcct caatgataga tgtggaagat aatatagcac agcctgcctt tgctggtgag 10560 tggtgattag gcctggtgga actgccatca ataaactaac tgtgattcag gtgaaaaaca 10620 ggaaagaagg aaatatgggg aaatagagtg aatgtcagat ggatcagaga gatacagtca 10680 tggggataag gtgtggtatc aggaataatg tgggaggcca gactgaagcc caggccaggt 10740 acaatggtaa ttgtgggaga ctcaacagag agtgggtatt gctaaaggag ctggggaaca 10800 gaaagtatat gcatcaggtg tgaggaagga aatatatttt ggaagttatg agaactgtat 10860 ggagtgagtt gagcatagtt tgtgattttg agggcttcta aaagtattag ggtggtggca 10920 gccactgcac agagacatga tgaccagcct aaaacagtaa ggtcaagttg tttggacaaa 10980 aaggctacag agcacggtcc cagtccttgt gtaagaattc cagctgcaca gccctgcact 11040 ttggctgtgt gtaatgaaaa gagttgggat gagtcaggaa gagctagtat gggaacagcc 11100 tctagagctg tcttcaagga atggaaagag gagtgaggaa aggatttagg acttatgggg 11160 tcagctaggt ttccctttgt gagtttatat aatggttttg ttaggatggc aaaaccaggt 11220 atccaaaggc aaaagtatcc aaccatgccc aggaaggaaa ggagttgttt tgtagaaggg 11280 gttggggttt cagagatcag ccagacatga tcagcaggga gaccccatgt gttttcatga 11340 agaattatgc tgaggtacgt aacggatgga gaagaaattt gagctttgga gggggatacc 11400 cgatatcctt tggagagtaa atgttgaagg agcaggaggg tatcttgttg aaaagactca 11460 aaggagggac tacagagtag aaggtcatca atatattgaa taatgtggga agcagaggga 11520 tgggaagaaa gtaaatcatg agagagagct tggctgaagt aatgagggct gtcccagaag 11580 ccttgtggca gtacagccca ggtaagctgc tgggactgat gggtgtcagg gtcagtttag 11640 gtaaaagcaa agagaggctg ggatgtgtgg tgcaggggaa tagtgaaaaa agcatcttta 11700 agatcaagaa tggaatagtg agttgtggag ggagatattg agaacaaaag agtgtacggg 11760 ttgggcacca cagggtggat aggaaaacaa ttttgttgat aaggcgcaga tcctgaacaa 11820 acctgtaaga cttgtctggt ttttagacag gtaagatggg ggaattataa ggagagtttg 11880 taggctttag aagcccatgc tgtagcaggt gagtgataac aggcttcaat ctccttaaag 11940 cctcttgtgg gatgggatac tggcgttaag ctgggtaagg gtgattaggc tttaatggga 12000 tagtaatggg catgtgatca gttgccaggg agggagtaga ggtgtcccat acctgtgggt 12060 taaggtgggg ggatacaaga ggaagacacg aaggaggctt tgggttggga agaaggatgg 12120 caatgagatg tagctgtagt ccaggaatag tcagagaagc agataatttg gttaaaatat 12180 ctcagcctaa taagggaact ggcaggtggg gataactaaa aaggagtgca taaaagaatg 12240 ttgcccaatt tggcaccaaa gttggggagt tttaagaggt ttagaaacct ggccgtcaat 12300 acccacaaca gttatggggg caagggaagc aggcccttga aaagaaggta gtgtggagtg 12360 ggtatccccc ctatcgatta cacaggggat ggacttaccc tctgctgtaa gagttacctg 12420 aagtttggca tccgtgattc tccggggggc ttccgaggtg atcgggtagc atcagtcttc 12 480 agccactaag ccaaggagat ctgggaagga gtcagccaag aaacattggg tttgagctcc 12540 aggaacttta ggagcagtgg cgatgtgagt cagacagtcc aacctctaat tggagcccac 12600 acagacaggg cacggcttag gaggaatccc aggctgtggg ccttctgagg cccagtggcc 12660 aggcttttgg catttgaagc aaggtccatg aggaggtttt gaaggagccc ctgggagctg 12720 tggcttggat gttctgaagg ttttgtatgc tggagacatg gtcttgggtt gtcttacact 12780 ggaggcaagt agctataatt cagagatgca ttgtcacttg gccgcctctt ctctattatg 12840 atacaccttg aaggtgaggc tatttaagtc ctgttgtggg gtttgaggcc tggaatccaa 12900 tttttggaga ttttttctaa tgtcaggaga ggattgggtc ataaaatgca tattaaggat 12960 aaggcggcct tctggcccct ctgggtccag ggctgtgaag catctaatgg taggtgctaa 13020 gtgggccatg aactgggctg ggttttcatc tttaccttgg gtagtttcct ttagcttgtc 13080 ataattaaca gccttgtatg ctgctttttt gaaccccttg actaggcagg agaccatgta 13140 atctcaccta gctatacctg gggagtccat cttgtactgc caatggggat gctcttgggg 13200 aactgccctg atgccttctt gaaggtctgg ctcatggcac tggtgggtgt cagtgtgaga 13260 ttgggctagg gtatcaactc tttcctgctc atctggggag aggggggagg tcaggatgac 13320 atttcagtca ctccaggtta aatcgcagga ctgagttaaa tattggaatt cctgtatata 13380 tttagtgggg tctgatgaaa aagagcctaa acgctgactg atctgggaga ggtcgataga 13440 gaaaaaggca catgtacctt gactatgcct tcagctccag ccacctgact aagagaaaat 13500 tgttgggcag gtggaggagg gctagtctcg gaatgaaact gtaaggtgga gtgggtgtga 13560 ggaggggagg tgatacttct attatagggt gggggagcag aggatgagga agaattggga 13620 cctggcttgg cctggtgagg agcagcctgg tggggagggg agaggtcaga tgggttcata 13680 gaaaaggagg attcaaagga ctcagagctt agggtggaga ctgaagaaag agacaggaga 13740 gaaagaagaa agatttggga tgagtcacac tgggagcaga gactagggag agactgatgt 13800 ataaaagaat gcctggatgt caggcacctc agaccatttg cccatttttc aacaaaaatc 13860 atccaggtct tataagatcg agaaatcaaa agtgtcattt tctggctatt tggaaccatt 13920 gtccagtttg tattgaggcc aagcggtatt gcagaagaaa ataaggcatt taggttttag 13980 gtcaggtgtt agtcaaaggg ggttttaggt tttgaagaac acaggctaag ggagaagaag 14040 agggagtgga ggctggaagg ttgcccatag tgaagggggc aagcccagag aaaagagaag 14100 gtagagacat ggagagaggg gagatggtac ttgccaccca gggaaggtgg tgcttgccac 14160 ccagggaagg tggtacttgc cacccaggga aggtgatact tgtgacccag gggaggtggt 14220 acttgtgcca aggtgaatga tcaaggcagg catccccaca gtgatcagat acctctgaaa 14280 tgtgggtgaa taaccaggca ggtgtcctgg cagtgattta gacaccaagg gaagactgtt 14340 cccgagtcca tgactggcac cagagttttg agttcatgga taaaacgtat ctcctccgtc 14400 tctaccagaa agggaaagaa actgaaatta aggaagggag agattgaagg gtggagggat 14460 agcaagagag gttggagaag agaataaaaa gaggccgctt acccgattta aaattggtga 14520 gatgaccttg ggctggctgg tctgaggacc caagaggtgg atctccccat ggagtgaggg 14580 cgaggacagg ggatgggact cccgaaggag tccccctgtc ccaggtcttt ggcaccaaat 14640 gtcatttgcg tccatgtgaa gagaccacca aacagtcttt gtgtgagcaa taaagctttt 14700 taatcacctg ggtgcaggct gactgagtcc gaaaaggagt ctgcaaaggg agataggggt 14760 gggtcagttt tataggactg gggtaagcag tggaaagttg cagttaaagg aagttatcta 14820 ttgtcagcag aggagggggt cacaaggtgc atggtaggga gatcataaga ctcattgtcc 14880 agaagaagaa tgtcacgagg tcgatcaatc gatcagttgg ggcagggcag taacaagtca 14940 taatggaacg ttgtaaggtt ggtcaatcag ttaagacagg agctggctgt ttcacctatt 15000 tgtagttttt ggttgcctca ggccatctgg atgtacccat gcaggcttgg gctaagaggc 15060 ctgaaaccca ccactttccc atgtcaaatc tttagtagat gtacccccaa gatacacatt 15120 cctcggacct tcttttccat agttaaaact tcacccctga aatgtagaaa caggaaggtt 15180 tttttttaag tttcagtgca aactcggagc aagtgtcata attttctgtc tccgatgtgt 15240 agaggtgaca ttttctcaga actttcatgt taagctggaa aactggaaag cgagtccact 15300 ttgtcattct gtcactcact cattttctca ctcaacaaac atgcctcaca cttatctaaa 15360 tctgctaggc taaaagaggt ccctggtgtc tgtaactttc taattctgct agaattctag 15420 agtgagctca tgaaataaat gaaaaggatg aagaacaaag agaaaaaaga ctgcacgttc 15 480 ccttctggcg ctcactcaca ttccctcagc ctcagtttct ccacatgccc ctagaggtga 15540 tcattcaagg atttatgaga ttttagagac aacacatgaa aaagcaaaga gacatcagaa 15600 agacaaggag ttacttagta tttatacaca aggataagac attcagtatc gacaacactt 15660 aaagaaaatt caagagtgat tttaaatttc ccatttcaaa tacctcctct attttgtttc 15720 ctttctctta tgacatctct aaataagctt cctctaactg ccagcaagtc taatttcatt 15780 ggatttgact gttttcaacc ccaattagag gcagggttaa gtacacttaa aataataatg 15840 aaatattatt ttgtttctcc tcccagggct attggacgtt tgacatggaa tgttataaaa 15900 agtatagaaa agtctggggg tgagtattct ggaagattcc attggataga cttgtttcta 15960 tgatgagttt accccactgc acagaggata gtctcagccc agagcctctt gcgatgaagc 16020 tcttgccagc ctaactacaa acagagaggt tttctgaaag aagaagatat gtatttggga 16080 gaagagtatt agaatgggaa actgtgtgcc attataagct atgtgcaaat tcagggaggt 16140 aaaggaagac aaagatgttc cacagaaaat atgaggagga cttcatgtgg attttgaaat 16200 agttctcctt cactacaaag atcagtaaca agggtgatgc cacaccaagg ttggacaggc 16260 agttgctggg caggtgtcct tgcagaaata tttttgtgta aggttgggtt gggctttctg 16320 caagctggtg gttttgccaa gtattttgtg atagttttgt tatcaggcac acaagcatga 16380 gaatccactc atcatagcct tctttgattt atttgtcaag gtttttaatc acacacatag 16440 gcacacagac acactgacac acacacacac acgtgtcata attttggttc tcacaatgtt 16500 cacactcgct attgtaaaac aactttttga gatttgtcct accaaggatc ccatatgtca 16560 ccaggtgttg aggtcttcgg tctaaactag gctgagacca ttgtggttac cacttttctc 16620 caagctttgg tggcccaggg actcccagca tcacgttctg tccagtgtct gcctattccc 16680 ctcttctttt tttcttccct tgatgccctt ttatcacatg cattgtctca gacccttcga 16740 atatgtgctc ataaatgcat ggcatcatct ccttcccaca tcgattcact ttcaattaaa 16800 agccaaaact ctttcatttc aactttggat ttaacatgct tttgaaagaa gggttgagaa 16860 atatagagaa acagattggg aaaccatgct ctgctgtttc ttttttttaa actttctatg 16920 taagtgtgga atttttcatt ctgttttatt attaacttta agccaagact ttttaataga 16980 aggatatata aatacatctt tgtctataca tttctgctga atttgaagag atgctgaata 17040 ttcttaaacc attgtgttcc ctggtgggct gatggactgt gattttataa ggtggcctca 17100 gccaactgca gcagctgttc cctgtcagag gggctagagg tttggcaaga gcggtggaag 17160 aggtgcagtg gtgtgttcgt tcactagaag cattagggag aaggttttgc ctgtttgtat 17220 ttcatcttct ctcatcaagt cctcagaaac cacagtgctg tctgcagggt gctgtggatc 17280 tggcatggcc catacaggca acatgactga gtagaaagga cacacagctc tggatgtcct 17340 tgggccccac agcaactgcc cttgaaacat ttagtccttg tgagcatttg atgatttact 17400 tgccttcaat tttccatgga cctaatactc tttataaagg gaaatatttt aaacctatga 17460 aacattgtgg agaatggcat gggaaatacc catgtatgca ccacccagct taacaaatgc 17520 tctcctgtca tttctaacca caatctcttt gaagagctct tttgtctttc aatctctctt 17580 ccctgtttgg cccacattac ccttcatcgt atgaagactt ggatggctcc tgtgtcagac 17640 tcttgctgtg agttatacct aatgaactag aacttaagat tactgtgtat tgtacatcta 17700 gaggattatg taatgtccgg atcaaagtct ggcttcctgg gtttggctcc agctgtagca 17760 taaggctgtt gatgtttaat caactctgtt ttctccacac agtatttatg actgtcaaca 17820 gcctatgctg gctatcacag atcccgacat gatcaaaaca gtgctagtga aagaatgtta 17880 ttctgtcttc acaaaccgga gggtaagcat ccattttttg aaattaaaat actcattgat 17940 tgattaaatt tttattttaa aattcttata tattcataga cagttgccta aaaactgtgc 18000 aggaaggttc catgtacact tcatcctgcc catcccccat ggtaacatct tgcaatcttg 18060 cataactata aatacagtat attcacatta ctgcagtaca ggaaattaac attgatacag 18120 ttcacagagt ttattcagat ttcaccagtt ttatgtgccc tcatttgtgt gggtagtttt 18180 ttgtttttgt ttttgtttgt tgtttgttgt ttgtttttcg gactgagtca cactctgtca 18240 cccaaatggg agtacagtgg tgtggtcttg gctcactgca acctccacct ccagggttca 18300 agtgattctt ctgcctccac ctcccgagta gctgggacta caagttgcac caccacgccc 18360 tactaatttt tgtattttta gtagagacga ggtttcacct tattggccag gctggtcttg 18420 agtgtgtggg tagttctatg caaatttatc acatttgtag atttgtgcaa tgaccagcat 18480 gatcaagatg cagacccatt ccatctccgt atggctccct gctactatcc tacagtcacg 18540 agtgtttttc tctgagaggg tctttatcag ggaaaacttt ttttattttg aggaggccaa 18600 tgtattaata tttttttatg gattgtgctt ttggtgtcaa gtctgaaagt tctttgccta 18660 gccctccttc ctatattgcc aatttttcta aaagtcataa agtttaatgc tttactaaat 18720 gtaactctac tatctatttt gtgttatttt ttgtataagg tatgagattt agatcaacgt 18780 tcattttttg tggcttgtag ttgtccagtt gttccaaagc catttttgga aatgtgggca 18840 tattgttaaa aattaattgg gcatgtttct ttgcatcttt catttgctat tctgtcccat 18900 tgatcactgt gtttattcct ccatcaacaa caaactggtg actctaatac ctgtacagta 18960 actcttagca tcatgtagag tgattcttcc cacttttatg gatgtatttt tttagatttg 19020 ttttagctct tcctggtcct ctgcttttcc ataaaaattc agaatgagtc tctcagtgtc 19080 tataaataaa catgctgtta ttttaacaag aattgaatta aatgtataga tcaatttagg 19140 gaaagtcttt atcttcacta tgtttggtct ttcaattcat gaacatagta tgcctctcca 19200 tttccttaga ttttctctga ttgcttttag caatttgtag ttttcagcat agagatcctg 19260 tgcatgtttt gttggattta cagctaaagt atttcatttt tgttggagtg attgtaaatg 19320 atattatgtt tttcttgtat tttcagatat tgattgttgg tacatagaaa catgcttggc 19380 ttttgtttgt tgatcttgta tcatagaacc ttgccgagct gacttagtat ttctagaagt 19440 ctttttgtat attcttgaga ttttatacat tgacaattat gtcacttgaa aatagagaca 19500 attcttttat ttcctttcca atctgtatgc cttttaattc tttttcccag tataatgtca 19560 aataaatgtg ttgaattttt gaattgttcc taatattagg aaggaagcat ttggtctttc 19620 atcatcaaga atgatttaga tgaagtgggt ttttttgtag attctcttta tcaaatggag 19680 gaattttctc tccctagttt gctgaatttt taacataaaa gagtactgta agtactcatt 19740 ataaaacaaa atatggctgt ggaagatgaa agagagtttc aagcatgctg gcttgatagg 19800 ccagatccaa gctggcaaaa ataattatct ctttcttctt tttttctatc catggaataa 19860 aaaattaaga ggaaagaatg ttaatagaat cgcattattt cttcaaaata cgttgtgagt 19920 tttaaaagta ttacctacct tttttattat acttttttta gggtacatgt gcacaacgtg 19980 caggtttgtt acatatgtat gcatgtgcca tgttggtgtg ctgtactcat taactcatca 20040 tttaacatta ggtataactc ctaatgctat ccttcccccc tctccccacc ccacaacagg 20100 ccccagtgtg tgatgttccc cttcctgtgt ccatgtgttc tcattgtata tttttttaaa 20160 tctaccacat caaggcacct ctttttcatg ttgcccatgg tttaggtgaa cataaagaca 20220 gagctcgtct gaggcaacat acagtccaac aaagccacct gcctctctgt ctccactctc 20280 tctctacact gcacgcgtgc taggtgttga tcctgtctat tccagtggaa gaacaggttc 20340 cgtaccatgt ggagaatttg catgtaaaag gagactggga tatacaggct ggagaccaca 20400 tcaggtggct gggcatgtgg gataaatcct attgagcatc tgtcataggg cctgtcactt 20460 agtagacagt cactaaatat ttgttaaata catgatgcct gtttaacaca ttttctacaa 20520 ccatggagac ctccacaact gatgtaggac aaaatctttc tgctttgaac tctagccttt 20580 cgggccagtg ggatttatga aaaatgccat ctctatagct gaggatgaag aatggaagag 20640 aatacgatca ttgctgtctc caacattcac cagcggaaaa ctcaaggagg tatgaaaata 20700 acttgggttt taattagaaa cttaaagaat gaatcaggtg gggacaggta gaaagtaaga 20760 tcagagttcc tttccgagga gtagtctgct gaatttgagc ttcctaaaaa tagtcttttt 20820 atgtacagaa aacacatcat aaaattcatt atacaatgtc acttattgtt ccatgccagg 20880 caaagtcatg tccttctggg acttatgtct gcacatttaa ctatgggtgg tgttgtgttt 20940 tgtgcttaga tggtccctat cattgcccag tatggagatg tgttggtgag aaatctgagg 21000 cgggaagcag agacaggcaa gcctgtcacc ttgaaacagt aagtaggagc acagccatgg 21060 ggttctgagc tgtcatgagc ccctccagct gcctgctatg gagctgatac tcccgctgtt 21120 gggttattcc agtgaccaga caaaaggagg gctgtggtaa tgcaacttca atgggtctcc 21180 caagatgggg cagctccgat gaggaggtgg ggcagctgga ggaaaaggat cttctcccct 21240 gtgcacagag gtcagggttt acatatctgt taaattgtca ccttggatat tctggaggac 21300 taaatacatc ctttaggggg aaaagtgtga ttgtatcaaa gttttaagca tggagtgtat 21360 gggatggtgg aaggggaagg cacttggtat ctgttggttg gcagtgagta gggtgggaaa 21420 gttataatgg agaacttaga ataactttga tcatttcatg ttttttttct gagggtatca 21480 gtagaatact aaatattaaa cattcccacc atttcttttt cctccagtct caaagagaga 21540 gggtggtaaa aacgctatag gtggggcaag cctattattt gctgtctaca cttatgcagg 21600 aacaacaggt gtaatctgag cctgtcctgg gcagacaggg gatatgtggt cactcactat 21660 agaagtttcc aaatcaaatt ttgagagttt tttttaacca ggacatcatt tgtcattata 21720 ttttacaaaa ataattctgc catcagggca acctcagctc accacagctg gggatagtgg 21780 aattttccaa agcttgagca gggagtatag agaataagga tgatatttct aggagctcag 21840 gacatggtac tgttgctttg taaagtgctg aagaggaatc ggctctgggc atagagtctg 21900 tagtcaggca atgtcacctg tcttgagccc cttagaaaga gtgaattttt ctactcttgt 21960 tctgctgaag cacagtgctt acccatcttg tatcatccac aattaacaca tgctactgca 22020 gttgtctgat agtggatctc tgtctttcta tgactaggct ccttgacctc agaggtaagt 22080 ctaactcagt tgagtgtctc catcaccccc agcggagagc cagctgtgtc actgacacct 22140 gataatcacc ttctgaggga gtgtgatggg agatgctcca gtaaatagtt ctgaaagtct 22200 gtggctgttt gtctgtcttg actggacatg tggatttcct gctgcacgca tagaggaagg 22260 atggtaaaga ggtgctgatt ttaattttcc acatctttct ccactcagcg tctttggggc 22320 ctacagcatg gatgtgatca ctagcacatc atttggagtg agcatcgact ctctcaacaa 22380 tccacaagac ccctttgtgg aaaacaccaa gaagctttta agatttaatc cattagatcc 22440 attcgttctc tcaataagta tgtggactac tatttccttt tctctccctc tctcttaaaa 22500 ataactgctt tattgagatg taaatcacca tgtaattcat ccacttaaaa tgtacagttt 22560 aatggtttgt agtacatttg aagatatgta cgaccatcac cattgtaaac tttaaaacac 22620 ttctgtcatt ctagaaacct catacatgtt tagctatcaa ccccctgcca gaaatcctgt 22680 tgtcgtatat aaccactagt caactttctg catctataga tttgcctatt ctggacactt 22740 catagaaatg atattaattt ttcagggttt ttttaattct ctagttcatg aatttgtact 22800 ttagtctgta ttattttctt tcttctgctg gcttcaggct tagtttgccc ttctccgttt 22860 actgtgttgt ggcatgaatg tagattattg atttgtgggt ttttttcctc taaatttaga 22920 cattacaact ataaatttcc ctctgggcac ttgtttgcta aataacatga cattctggcc 22980 tatcacatct taatttttgc tcagctcaaa acagtttcta tttgcccttt taatttctac 23040 ttcaacttat tggttgctta aaagtttatt gtttaacttc cacgtatgtg tgagtttccc 23100 aattttcgtt accttattga ttttagcttt atcctatgat aggtgacaga gatacactgt 23160 gttatttcta tctgatttct tgaagagcat gattaattta agcagcaaat tatgatttgg 23220 gttatcctaa gagaccctag cccaatacat aaaggcaaag agattagggc attgaatttt 23280 gttcctttca tcattcaaaa gatgcagaag gggctgctga tctcactgct gtagtggtgg 23340 tccctatgca cagaccttcc cttgctcagc cactggcctg gaagaggggc aaaagtcata 23400 gaaggaatgg cttccagttg agaacactga tgtatttcac tcttctgatt ggtagagaaa 23460 attagaattg ctccaggtaa attttgtatg ttcacaatgg tttcgttttc tgttggtttt 23520 ttttttctac agaagtcttt ccattcctta ccccaattct tgaagcatta aatatcactg 23580 tgtttccaag aaaagttata agttttctaa caaaatctgt aaaacagata aaagaaggtc 23640 gcctcaaaga gacacaaaag gtaaaacgtg gtggtagtta taggaggatg tttagttttc 23700 ataatttttt agaaatatac atatgatcgg tgcacttccc tgtatgtttt tagagaaaga 23760 ttttaacata aagagtactt atgtttagag caagataatt catttggtag gaatgcagaa 23820 agtggagttt gaggaaggag atatgagaat gagtcagaga gagcacttaa aatttgatat 23880 caggcacaag aattagtatg ccatgacata aatattattc agataaaacc ttatcacagc 23940 acttcccttc aataaattgt caaaggataa acattactgt ttggaaatac attttattgg 24000 tattatgctt tctctagatc atagattggt aaagagtcat ttcaagtaca aggctattat 24060 tttacacatt ctccaacaaa aggtcctatc aggccacctg ctgaagtgta gtgtgtgtta 24120 catttgagcc ctctgacagc ctcgagtgaa cagaggtctt ctctgggaat tgaggctgca 24180 aggctggcta actcagcttt gccttcatga gccctggagt ccagccaaaa gatatctgca 24240 ggtcaggggg gcaagaaggg gtaacccagg tgtcactgaa gattgtacag atttggggaa 24300 tgttgcagta tttgaaagtg ctcccaccaa agctgctgac aagttctgga aatgtcagga 24360 gaataatctt tgtggacatt ggtgatgaag aagacagaag aataaaaggc ccaggacttc 24420 ttcatgggta tgcagttagg accagggaaa agaccaaaaa gcaattgaac attttagcca 24 480 gatttgtttc aggaaaacat cctgctttcc aaggatttag atggatattc cttttcactg 24540 gtgactcagg caacacccat cagatttctg gtctccaagg ggctatattc tcagaaggga 24600 ggttgaggac tgcacttctg attaacttct gacttcacga gtcactttct gtcaataaaa 24660 tttctctttt tgcttctagc accgagtgga tttccttcag ctgatgattg actctcagaa 24720 ttcaaaagac tctgagaccc acaaaggtaa caaaagagtg cttctgaggg ctactggtgg 24780 ggcactcaga gggaaggttt tgttctgaaa atgtgcagga agtattccag gtagatgaga 24840 atttctgcca catagcagaa tgacacaggt ttggatgtta aaatggtagc tggaggcaac 24900 ttctagaggc atgcaggcat agatagccat gttctaagct taaagggcaa ccctaaggaa 24960 aactggcatg ccaggaagag tcagcagtct gtggatcacc tacataagat caaatgccag 25020 ttctcagcct cctccagatc cacagaatta aagtttcttc ctggaaattt attaggtttc 25080 tgcaaaaata attgcaattt ttgtgattgc tttcaatggc aaaaaccgca attacttttg 25140 cagcaactta atgcatctaa cacctaatca ggagccatgc tatccaaata agggcgattt 25200 taactggcaa tttttggtga aagtgtgttc accaggttaa tcaactatga agttattttt 25260 tcctttgcaa tggataagta actttttgga aaacctctga gactgtgtaa atatcctttg 25320 tcttcagtct ttcacctacc agttttacaa cccatgtacg atttttactg aagtcattgt 25380 tactatgagt ggcaaaatga tggccatgtt cagtttcaag ccaccaagat ttggcaatca 25440 tgtcacaaaa ttcctgaata tttaatcatt ggttctagag agcaggagta ggctgacttt 25500 ggcatacaga tttaaatatg tccatgtcta catctacttc tgtctgtctg tctatctatc 25560 tatcatctgt ccatcaatca tctatttatc tacttactga ttatctagct atcatctctc 25620 tatctatccc aatacaactt gctgtgataa aggaaatagc ctactgtttt agtgtttcat 25680 ctagaaatca ctagacacat atagctattg agcactgcac atgtggctag tgccactgat 25740 gaaaaatttt aagttgtatt taagtttaat tgaataaaat ttgattttaa atagccacat 25800 gtcaatagtg gctaccacat tggacagcag agctctaaac tttgagatta tagtttggtt 25860 tcacatcagt atcatcaggt tcatggtaac agaatacaat tattagatag ttgaatttac 25920 ccacaactgc atcacagaaa tcttcactgg taaatcacag ctctgttgac atttctcaca 25980 ctcctttcgt attggttttg gtggtgagtt acatggttgg aacaggcact atatttattt 26040 ctgtgttccc ggtctctaaa tgtcctaatc cagtcctgat caaacagacc agtgatgggc 26100 catcctgagc ttctctcagg aaaaaaatca agagaggggc caacttgcaa tcacaagagc 26160 ttataccatt ttaatgccat tcatcagact gcaatcaatc accactcatc tagctttttg 26220 tccatcttca gttgctgtac atcctgagat agtcaattct gagacctata gcctgggtct 26280 attacctggt gcctctcaaa gatataatcc atacttctca acctgggcta tgcacacaaa 26340 ttaccacatc ttgtgaaatt cagacttgaa tcagtagttc aatggtgggg tttgagattt 26400 tgcatttctg tttttgttgt tgttgttgtt gttttgtttt gttttgagac agagttttcg 26460 ctcttgttgc ccaggctgga gtgcaatggc atgatctcag ctcaccacaa cctcagcctc 26520 ccagattcaa gcgattattc tgcctcagcc tcccaagtag ctgggattac aggcatgcgt 26580 caccatgcct gggtaatttt gtatttttag tagagaaggg gtttctccat gttggttagg 26640 ctggtcttga actcctgacc tcgggtgatc cacccacctt ggcctcccaa agtgatggga 26700 ttacaggtgt gtgccaccat gcccagcctt tgcatttcta ataaggtcac agatgcttat 26760 gatccacgga ccacactttg agtagcaaga ggtggtctgt attccaatat tggtctctta 26820 agttccctca aacatatcta aaggaaaggt cctttaattt tccatacatc tagccattaa 26880 tgttgcaaat tctctcaaag tttgtcaagg tatattgtag ctaaaatgaa ttacattttt 26940 cttgggagag aagactaccc catattaatt tacaataaag tacttttagg atcattcaag 27000 gagcacacca tgacactgag tatgttatgc agaaatgttc tctctggaaa ttacccagct 27060 gtgaaggtgg cagggatggc acgaggagga gcggatggtc tacgttcttg aagacattgg 27120 gggaaaactg gattaaaatg atttgccttg ttctggctct gtaagacaca catcagagtg 27180 aaaccacccc cagtgagact ctgaattgct tttctatttt ttcccctggg gatttgaagg 27240 ttccacttag atttcttttc atctaaactg tgatgcccta cattgacctg atttacctaa 27300 aatgtctttt ctctcctttc agctctgtct gatctggagc tcatggccca atcaattatc 27360 tttatttttg ctggctatga aaccacgagc agtgttctct ccttcattat atatgaactg 27420 gccactcacc ctgatgtcca gcagaaagtg cagaaggaaa ttgatacagt tttacccaat 27480 aaggtgagtg gacagtacat ggagaaggag ggaggaggtg aagccttagc aaagatgctt 27540 cctcaccact tcccaggaga atttttataa aaagcataat catgattgtt tcattcacaa 27600 tatgtaggaa ggctctcaaa ataaaaagaa ggaagcatag ggaatggttg ctattggcag 27660 aagtataaga tctttgtaaa acagtcctct ccctggttca tctgctttct gttaccacaa 27720 taatgctaag taaaaaaaca tccaaaaacc tccctggcat ttaacaatat gcatattgct 27780 cacacgtctt gatagtgagc tctgctgata ttggcaggac ttgctctggt ctggctgtga 27840 tctgatggag cctggccctg ggtgcgctgt gcaggttgac tcagctctgc cccacatgtg 27900 tctcatgttt cagtcaggta accactggtg aagaagcaag ctaggaacca gggtatctga 27960 cttctgagct aaactcttaa actctataat attgcctttc aaatataaca ctaagtacta 28020 ggtgcctatc aaccacactg ttttcagacc tctgccaaaa cttggattct ttgtgctatg 28080 aagagacatg gcttttgggc tggtctttgt gtggcagtga ggtgagcaca aagggatgtt 28140 cttcagagat tacagtccag ccctgaagca acaactagga gactgtttca gcaagtgagg 28200 acagggctgt gtggggttct atccttttca taactttgcc tggcactgaa atcacatgct 28260 ctgataacat ccaccagaac tttcttttgt catatttggg atagaaaggg actagttttt 28320 cctcaaatta ttgatagaga ttttatataa tatagtgttt ctctccacat tttatgtata 28380 taacaaaagc cctgcttttg tgtatatatg catatatata tatatacaca cacacacaca 28440 tatatataat acaaatcctg ctttgtaact gtttttgttt gtatatataa caaaaagagt 28500 tatgaaccag aagtttggcc aataatcctt gtcgcacaga gaatttgctt tttctatctg 28560 ttttcacttc cttggttaca gacgtgtaac ctcttttttg aatggtgaca atcactttgt 28620 catattttat ttgatgctag tggtcatagc ctattagtca tgtttgcttc catgagaaag 28680 aaaaaccact acatggttat gctaaggatt tcagtcattg gggttagagc cttcccgaat 28740 gtctcctgct ttcataactc ctccacacat cttagtgggc cattgagcac atcaaagggc 28800 atgacagtta ttaaaatact ttatgaatgc tacaatcctt tgccagtatg agttgttctc 28860 tggaacttct aacagttcaa cagtactaca tggactgagt taaaagttaa ttcaaaaatc 28920 tcaatttatc caaatctgtt tctttctttt caggcaccac ccacctatga tactgtgcta 28980 cagttggagt atcttgacat ggtggtgaat gaaacactca gattattccc agttgctatg 29040 agacttgaga gggtctgcaa aaaagatgtt gaaatcaatg ggatgtttat tcccaaaggg 29100 gtggtggtga tgattccaag ctatgttctt catcatgacc caaagtactg gagagagcct 29160 gagaagttcc tccctgaaag gtaggaggcc cctgggaagg gagccctccc tgaaccagcc 29220 tggttcaagc atattctgcc tctctacagg acagtctggg cttgtacaat cgtttgcttg 29280 tctttttatg tttaaaaggt tttttcaaat catgaaattg atcattgtca cactttacaa 29340 accacagact agataaaaga aaactatagc cagtcacagt cccagcaact taagatgaag 29400 gtcctcaatt atgtccttat gggtcataag tgtccaaaat gtaaggactc ttttaaaaac 29460 acatgatcac aatgctatta ttatgtccca caaatgaata ttttttcctg aatatactca 29520 aatcttcagg aatcaaattt gaataaaaaa catgcgtcta atcttcaaag aatttatagg 29580 ttagtgcaac agatagacaa agaaagcagt gatgacactg ctttccatca atacagtagc 29640 atcatatgcc tgtgtaaatt atctgactta aactattcta tggaggtgtg ggggagaaag 29700 aaggagagat ggagattaga agaaggagga gaaggaggag agaaggaggg gtaagacaag 29760 gtagggagga gaaggaggag aattagaaaa acaagagacg agaggagaag gaaagtgcaa 29820 aataacaatt ttgaagtagt gcaagacaat ttcttctcct tcctcatgac caacataagg 29880 gtgacttgag gcaggaatct acttttctgt cagtcattct catcacttat gtgccttttg 29940 tagtgtgaac acatcaccat cctgactata atttgagtgt ttagaaataa atatactttg 30000 caacagtatt tatctcctct caacaagact gaaagctcct ataatgtaag gagagtagaa 30060 aggatctgta ccttacaatt ctcatagcaa aatatgcata gcaggatttc agtgactagc 30120 ccacaaaagt atcctgtgta ctgctagtag aggggtgggc cctaagtaag aaaccctaac 30180 atgtaactct tagaggtatt atgtctttaa cttttaaaat atctaccaat atggaaccag 30240 gttcagtaaa aagaacaagg acaacataga tccttacata tacacaccct ttggaagtgg 30300 acccagaaac tgcattggca tgaggtttgc tctcgtgaac atgaaacttg ctctagtcag 30360 agtccttcag aacttctcct tcaaaccttg taaagaaaca caggtcagtc aattttctgc 30420 attaataatg ttttattaac aattatttta actgaatggt ctatatattt aaaaaagaat 30 480 acactcactt aatcttttaa taatttgttc tatgggccaa ggaatctatt tggacccatc 30540 tatgatcttt aagggtgctt cagttctgga gttcaaaagc tgtagcatta aaaacatcat 30600 gcaatgtcaa tgtagactag catgacatga ttatctacag tctccttgaa cttgagcaaa 30660 attaaattca gtttcaagtc aattggaaag atggggaagt attttgcaca atcatgaggt 30720 gcacttatta tcttgttttg acttttgaat gatgctcttc cttcccagaa cttgtagaag 30780 ttttctcatg acagtggctc ccaaccacta gctgtacatt ggaatcacca gggagcttta 30840 aaaattcatg atgcctggaa catctcagaa attctaaact aatttgccca gagtgtggct 30900 ttaaaagctc ccccattgct tctcatgtga agccaaggtg gaaaatgact aatttaaggt 30960 agttctattg gatacgaagg actaccacag tccaaggcca tccttactga cctcaccttc 31020 caggtgccta gctccaacca actgggctcc ttttcaactc aattataact ctgctaatct 31080 tctttccttt cttctcctca ttctgttctt atctttcaag tcctagttca attcccaagc 31140 ccctccaagg tgtcttcttg acctagtcca gggctcattt actcctctgc tctgttatta 31200 gttactggag gctattatca tataatttca taatttgcca ttaaatcatt gagttttgtt 31260 ctctatattt tctctttatc taagaatgtc ttcccccctc cattaacaat atcttctcat 31320 tttattccat ttaaaatatc ccagtggtgc cttgcaagtg acctcatcta actcaagcat 31380 ttggtcattt ggtaaatgtt taaggagtga tatgcagtcg attggtttgc atacaaatat 31440 taagtttttt aatgtgaaca tttagcaaat gacattcatg tatttgcatg tgtgtgtgct 31500 tgcacatgtg cacatgcatg tctgtctgca gggaaaatat attcatgcct tttgaaaatt 31560 tttaaataat gtgttatatt tatagaaaga tttggaacct tttctctgaa gaagttaaag 31620 aacagatgtc attgattcat attaagcaag accctataaa tcttatttct aggtctcatg 31680 tatttattaa gcaactccac accttaagca ggctttctac atagaagagg aagaagatag 31740 agatggtttc catattattt tcatattcca cattatttgt ggctttaggc cagctatgta 31800 gctatcctgt atgtgtgctc agacaggaga ctcagccctg agagaaggcg gtcctctggc 31860 acacctagga tggggaaggt actcccttgg aagtcccaag ctggcacttc tggatctcca 31920 tggcaatttt cttgcccatc actccatgga gatcagaata tcactctatt gtgtcccctc 31980 aacactgaag gagtgtctca ataagaaaag ttgagtcaaa acactgtagg aattgagagg 32040 ttccccactt gcactaccct tgtaaaccaa gagaagatgt taaaaaataa aacgataatg 32100 cttcctgaag gtgtcttccc atctttacac tagatgggtt caattgagag gaattactgg 32160 actgtggaag ttgaagactg tccacataat taaaatgtac aatagctact caggattacc 32220 ttgcaagttt caacatacac aaaattaact tcataagatg gtttaaaaag tttaccgtta 32280 tacctaataa tctggtttaa atttttaaaa ctcatccatt ttcgttaaaa tttaaatcaa 32340 aaaagaacac gggttcccat gaatttgtct caggtcaaac ctcacacaga ataggtgctc 32400 catgaatatt ttgttaaatg atagatgatg aatgttctca ctatccaatc ttcacacatc 32460 ttatagagta agtataacga atccaagatt tatagtgctg aaagtagttt ttatatgttt 32520 acaaagcatt attgtcagta attttttttt actttgatgc tatactttct acttttgctt 32580 tatttaatgc ttctcaatat gctcgtttaa ctgttgcaga tccccctgaa attacgcttt 32640 ggaggacttc ttctaacaga aaaacccatt gttctaaagg ctgagtcaag ggatgagacc 32700 gtaagtggag cctgatttcc ctaaggactt ctggtttgct cttcaagaaa gctgtgcccc 32760 agaacaccag agacctcaaa ttactttaca aatagaaccc tgaaatgaag acgggcttca 32820 tccaatgtgc tgcataaata atcggggatt ctgtacgtgc attgtgctct ctcatggtct 32880 gtatagagtg ttatacttgg taatatagag gagatgacca aatcagtgct ggggaagtag 32940 atttggcttc tctgcttctc ataggactat ctccaccacc cccagttagc accattaact 33000 cctcctgagc tctgataaca taattaacat ttctcaataa tttcaaccac aatcattaat 33060 aaaaatagga attattttga tggctctaac agtgacattt atatcatgtg ttatatctgt 33120 agtattctat agtaagcttt atattaagca aatcaataaa aacctcttta caaaagtatt 33180 attggatgtt tcctgcacat taaggagaaa tctatagaac tgaatg 33226

Claims (3)

A gene as a marker associated with dysfunction of the CYP3A7 protein, which encodes a variant CYP3A7 protein which generates a translation termination codon (TAA) at the 55th amino acid position in the amino acid sequence of SEQ ID NO: 21. The gene according to claim 1, wherein a thymine (T) base is inserted between a 134th base (C) and a 135th base (T) in the nucleotide sequence of SEQ ID NO: 20. A method for detecting a gene as a marker associated with dysfunction of the CYP3A7 protein comprising the following steps: (1) isolating DNA from a blood sample of the subject; (2) amplifying the separated DNA by performing PCR using primers of SEQ ID NO: 17 and SEQ ID NO: 18; And (3) confirming the presence or absence of mutation of the CYP3A7 gene by pyrosequencing method using the primer of SEQ ID NO: 19 as a template of the PCR product of step (2).