CN101864423A - MUC13 gene determining piglet F4ac diarrhea susceptibility/resistance and its application - Google Patents

Abstract

The invention discloses a MUC13 gene that determines the susceptibility/resistance of F4ac diarrhea in piglets and its application in genetic improvement of breeding pigs. The population, through genome-wide scanning linkage mapping, high-density SNP multi-point linkage fine mapping in the target region, breakpoint recombination analysis, linkage disequilibrium-based distant population association analysis, and finally locked the target gene encoding ETEC F4ac receptor - MUC13 gene, and isolated and cloned the MUC13 gene. Five key mutation sites were identified in the MUC13 gene. These five mutation sites were basically co-segregated with the ETEC F4ac in vitro adhesion phenotype in 144 purebred Du, Long and Big piglets from core populations in 5 provinces, and the strongest correlated markers determined the accuracy of resistant/susceptible individuals. The resistance reached 97% (P=1.59×10 ^-21 ). The invention provides a new method for disease-resistant breeding of swine ETEC F4ac diarrhea disease.

Description

MUC13 gene determining piglet F4ac diarrhea susceptibility/resistance and its application

technical field

The invention relates to the field of animal genetics and breeding, in particular to a MUC13 gene determining the susceptibility/resistance of ETEC F4ac diarrhea in piglets and its application in genetic improvement of breeding pigs.

Background technique

Piglet diarrhea is a common infectious disease in pig production, which has caused huge economic losses to the world's pig industry. Enterotoxigenic Escherichia coli F4 (Enterotoxigenic Escherichia coli F4, ETEC F4) is the most important pathogen causing diarrheal disease in newborn and pre-weaned piglets. Taking my country and Denmark as examples, about 35% and 40% of piglet diarrhea are caused by ETEC F4 infection respectively. In my country, its fatality rate is between 10%-30%, and even higher than this ratio in some places, resulting in a serious decline in the slaughter rate of pigs, the quality of commercial pigs, and the loss of economic benefits.

The pathogenicity of ETEC F4 depends on whether it specifically binds to receptors on the brush border of porcine intestinal epithelial cells. After this combination occurs, ETEC F4 proliferates to produce enterotoxin, which leads to the hypersecretion of intestinal epithelial cells, and a large amount of water and electrolytes enter the intestinal lumen, causing a large amount of fluid accumulation in the intestinal lumen, which exceeds the reabsorption capacity of the intestinal wall and causes diarrhea. Therefore, the presence or absence of ETECF4 receptors in piglet intestinal epithelial cells is the key to whether piglets are infected with ETEC F4 or not. Pigs without recipients were resistant, while pigs with recipients were susceptible. Isolation and identification of the porcine ETEC F4 receptor gene and its resistance loci are therefore the key to breeding for porcine ETEC F4 diarrhea resistance.

Existing studies have shown that the pig ETEC F4 receptor is controlled by a single gene, showing a recessive inheritance pattern. Pigs carrying the dominant gene S were susceptible to ETEC F4 adhesion, while pigs homozygous for the recessive gene s were resistant. Common Western commercial pig breeds, such as Large White, Landrace and Duroc, have a certain proportion of susceptible individuals. The susceptibility of ETEC can be judged by the in vitro adhesion test of the brush border of the small intestine. The adherent individuals are susceptible, while the non-adherent individuals are resistant. ETEC F4 is divided into ab, ac and ad three subtypes according to immunological characteristics, among which F4ac is the most important pathogenic bacteria in western pig herds. Therefore, the localization and identification of the F4ac receptor gene has attracted the attention of many research groups at home and abroad.

小组联合开展的基因定位研究中得到了验证，他们还利用FISH和辐射杂种细胞克隆板技术揭示ETEC F4ac受体位于13号染色体q41区(SSC13q41)，分别与人3号染色体的q28-29和q21区同源。2009年，瑞典LeifAndersson小组、丹麦小组和瑞士Peter Vogeli小组联合开展的精细定位将ETEC F4ac受体基因进一步缩小至SSC13q41的SW207与S0075之间。并提出该受体基因最有可能位于Sw207与MUC4之间的14Mb区域。In 1995, the Swedish Leif Andersson group used the European wild boar × Large White pig resource family group as materials, and after three-generation pedigree linkage analysis, for the first time located the ETEC F4ac receptor on chromosome 13, which was closely linked with the Tf gene, and found that F4ab and F4ac are receptors The body sites are tightly linked and it is thought that the two receptors are controlled by the same receptor. In 2002, Peter Vogeli’s group in Switzerland used Large White pigs and three-generation Large White × Changbai families as materials, and used more microsatellite marker information on chromosome 13 to further locate the ETEC F4ac receptor between S0068 and Sw1030, and S0075 and Sw1030. Sw225 is highly linked (θ = 0.00). This result was confirmed a year later by the Swedish LeifAndersson group and the Danish

It was verified in the gene mapping study jointly carried out by the group. They also used FISH and radiation hybrid cell clone plate technology to reveal that the ETEC F4ac receptor is located in the q41 region of chromosome 13 (SSC13q41), which is related to q28-29 and q21 of human chromosome 3, respectively. District homology. In 2009, Sweden LeifAndersson group, Denmark The fine mapping jointly carried out by the group and Peter Vogeli's group in Switzerland further narrowed down the ETEC F4ac receptor gene to between SW207 and S0075 of SSC13q41. And it is proposed that the receptor gene is most likely located in the 14Mb region between Sw207 and MUC4.

小组在欧洲野猪×大白猪资源家系群体中，发现了Mucin 4基因内含子7的一个SNP标记与ETEC F4ac的黏附表型共分离，由此申请了一项国际发明专利。申请人就该位点的影响效应在自身构建的白色杜洛克×二花脸资源家系中开展了研究，发现该标记在始祖动物中没有多态性，而该资源家系的F₂个体中有着充分分离的ETEC F4ac的黏附表型，这表明该SNP标记只是与ETEC F4ac受体基因因果突变(causative mutation)呈一定连锁不平衡的分子标记，不能解释所有群体的ETEC F4ac受体遗传基础。因此，尽管国内外其他研究小组在ETEC F4ac受体基因鉴别方面取得了一定的进展，但仍未有重大的突破，迄今没有鉴别到真正的ETEC F4ac受体基因及其关键变异位点。Identification of positional candidate genes in the SSC13q41 region is the focus of current ETEC F4ac receptor research. A large number of studies on immunobiochemical properties have shown that mucin-type sialoglycoprotein (IMTGP), 74kDa transferrin (GV74) and small intestinal neutral glycosylate H (mLad) conform to the physical and chemical characteristics of ETEC F4ac receptors, among which mucin-type sialoglycoprotein The glycoprotein binds to F4ab and F4ac, but not to F4ad, and the existence of this glycoprotein is highly correlated with the sensitivity of newborn piglets to ETEC F4ab or F4ac, so it becomes an important basis for the selection of ETEC F4ac receptor candidate genes. After obtaining the preliminary chromosomal location information of the ETEC F4ac receptor gene, research groups at home and abroad selected the adhesin glycoprotein coding gene (MUC gene family) and the transferrin receptor gene (TFRC) for research. Peter Vogeli's group in Switzerland Four genes, TFRC, B3GnT5, B4GALT4, and B4GALT4 were investigated, but no candidate genes and markers were found to be significantly associated with the F4ac adhesion phenotype; Denmark

The team found a SNP marker in intron 7 of the Mucin 4 gene co-segregated with the adhesion phenotype of ETEC F4ac in the European wild boar × Large White pig resource family population, and applied for an international invention patent. The applicant conducted research on the effect of this site in the white Duroc × Erhualian resource family constructed by the applicant, and found that there was no polymorphism of this marker in the ancestor animal, but there was sufficient segregation in the F ₂ individuals of the resource family The adhesion phenotype of ETEC F4ac indicates that the SNP marker is only a molecular marker in linkage disequilibrium with the causative mutation of the ETEC F4ac receptor gene, and cannot explain the genetic basis of ETEC F4ac receptor in all populations. Therefore, although other research groups at home and abroad have made some progress in the identification of the ETEC F4ac receptor gene, there is still no major breakthrough. So far, no real ETEC F4ac receptor gene and its key mutation sites have been identified.

Contents of the invention

The first object of the present invention is to provide a MUC13 gene and key marker sites thereof that determine the susceptibility/resistance to ETEC F4ac diarrhea of newborn and pre-weaned piglets; Assisted selection (MAS) selection of individuals with favorable alleles for breeding can significantly improve the resistance to diarrheal diseases of newborn and pre-weaned piglets in the population, and greatly reduce piglet mortality.

The second object of the present invention is the application of the MUC13 gene key marker for determining the susceptibility/resistance of piglet F4ac diarrhea to the genetic improvement of piglet resistance to diarrhea.

First purpose of the present invention is achieved like this:

1. Experimental animals and susceptibility/resistance phenotype determination

1.1 Experimental animals

The pedigree animals used in the present invention for target gene mapping all come from the resource pedigree group of white Duroc×Erhualian pig constructed by the applicant. In this resource family, 2 white Duroc boars and 17 Erhualian sows were used as progenitors to breed the F ₁ generation, and 59 F ₁ sows were self-crossed with 9 F ₁ boars to produce the F ₂ generation. Then select 62 F ₂ boars and 149 F ₂ sows to self-cross to generate F ₃ generation individuals. F ₂ /F ₃ population individuals of each resource family were slaughtered at 240±3 days old.

The remote population used for the fine positioning of the target gene used in the present invention is 292 purebred piglets, of which 12 local pigs representing 6 ecological types in China (including South China, Central China, North China, Jianghai, Southwest, Plateau, etc.) Breeds (including Erhualian, Jinhua pig, Jiangquhai pig, Shaziling pig, Yushan black pig, Hangzhou pig, Tongcheng pig, Lantang pig, Bamaxiang pig, Laiwu pig, Rongchang pig, Tibetan pig) 148 heads, each Select at least 3 paternal lines for pig breeds, and select 3-4 individuals aged 6-8 weeks for each family; in addition, purchase 144 Duroc, Landrace, and Large White pigs from large-scale commercial pig farms in 5 different provinces, and each pig The species also selects at least 3 paternal families, and each family selects 3-4 6-8 week-old individuals.

1.2 Susceptibility/resistance phenotype determination

After the pigs were slaughtered, the jejunum part of the small intestine was taken, and the brush border of small intestinal epithelial cells was extracted, and the E.coliF4ac strain was used to adhere to the brush border of pig small intestinal epithelial cells, and the results of bacterial adhesion were detected by phase contrast microscopy. More than 20 brush borders were detected for each sample. If at least 2 bacteria adhered to more than 10% of the brush borders, the sample was judged as adherent type. If at least 2 bacteria adhered to the brush borders, less than 10 %, but there are more than 10% of brush borders with 1-2 bacterial adhesion, the sample is judged as weak adhesion type, otherwise it is judged as non-adhesion type. The adhesion effect diagram is shown in Figure 1.

2. Preliminary mapping of the ETEC F4ac receptor gene by genome-wide scanning

The present invention firstly selects 151 microsatellite markers published on the website of the American Meat and Livestock Center (MARC) covering 19 chromosomes, 200 F ₂ generation individuals and all F Whole-genome scans were performed in ₀ and F ₁ individuals. details as follows:

2.1 Microsatellite marker PCR amplification conditions

Microsatellite marker polymerase chain reaction (PCR) reaction system is 15 μl, which contains 40ng genomic DNA (deoxyribonucleic acid), 0.2mM dNTPs (four deoxynucleotide substrates for synthetic DNA), 0.2μM primers, 1.5mM Mg ²⁺ , 1×PCR buffer (buffer) and 1U DNA polymerase (Taq enzyme). The PCR amplification program was 94.0°C for 4min; 40 cycles (94.0°C for 30sec, 55-65°C for 20sec, 72.0°C for 30sec); 72.0°C for 5min.

2.2 Electrophoresis and analysis of microsatellite marker amplification products

-400HD ROX以100∶1比例混合而成)，进行混合、振荡和离心后，上样于ABI3130XL自动遗传分析仪(3130XL Genetic Analyzer，ABI公司，美国)进行毛细管电泳。经美国ABI公司提供的公用的DataCollection Software V.30收集PCR产物电泳检测后的原始荧光信号，采用美国ABI公司提供的公用的Genemapper3.7读取微卫星片段的大小，并校正错误数据后，将Excel格式的数据转换成CRIMAP(2.4)软件要求的数据格式，再次检验所有个体的基因型。根据家系记载，确保所有个体的数据符合家系分离规律。然后利用公用的CRIMAP软件进行连锁分析，结果如图2所示。从图中可知，ETEC F4ac受体基因被初步定位于猪13号染色体(SSC13)的SW207与S0075之间，这个区域大约17cM。After the PCR conditions were optimized, microsatellite polymorphisms were detected using the _F1 individual genomic DNA of the resource family, and the combination of microsatellites was optimized according to factors such as microsatellite primer fluorescent marker color and fragment size to improve the efficiency of capillary electrophoresis. After PCR amplification, the product was first electrophoresed with 2% agarose gel, and then the quality of the PCR product was detected by gel imaging with a gel imaging system to determine the dilution factor of the PCR product, and then the PCR product was diluted in an appropriate amount to take 1 μl of combined diluted product was added to 10 μl of buffer (purchased from Hi-Di ^TM deionized formamide and

-400HD ROX at a ratio of 100:1), after mixing, shaking and centrifugation, the sample was loaded on an ABI3130XL automatic genetic analyzer (3130XL Genetic Analyzer, ABI Company, USA) for capillary electrophoresis. The original fluorescent signal after the electrophoresis detection of the PCR product was collected by the public DataCollection Software V.30 provided by the American ABI company, and the public Genemapper3.7 provided by the American ABI company was used to read the size of the microsatellite fragment, and after correcting the wrong data, the The data in Excel format was converted into the data format required by CRIMAP (2.4) software, and the genotypes of all individuals were checked again. According to the family records, ensure that the data of all individuals conform to the law of family separation. Then, the public CRIMAP software was used for linkage analysis, and the results are shown in Figure 2. It can be seen from the figure that the ETEC F4ac receptor gene is preliminarily located between SW207 and S0075 of pig chromosome 13 (SSC13), and this region is about 17cM.

3. Fine mapping of ETEC F4ac receptor gene in the target region

The applicant has added molecular markers to 51 (including 33 microsatellites and 18 single nucleotide polymorphism markers) in the area (SW207-S0075) of the ETEC F4ac receptor site and its two flanks. See Table 1 for marker primer information. These markers were genotyped in 755 F ₂ individuals and all F ₀ and F ₁ individuals in the white Duroc × Erhualian resource family population. The 33 microsatellite markers were detected in the same way as the microsatellite markers in the genome-wide scan described above. The detection method of 18 single nucleotide polymorphism markers (SNP) is carried out by the SNaPshot method, as follows:

3.1 PCR amplification conditions for SNP markers

In a 25 μl PCR reaction system, 25 ng of porcine genomic DNA, 1.0 mM MgCl ₂ , 100 mM dNTP, 10 pmol of forward and reverse primers, 2 units of Taq enzyme and 1×PCR buffer (Shanghai Bioengineering Company, China) were included. Amplification conditions were: 94°C for 4 minutes; 94°C for 30s, 56-65°C for 30s, 72°C for 30s, a total of 35 cycles; finally extended at 72°C for 10 minutes. PCR products were detected by 2% agarose gel electrophoresis and photographed. Take 3 μl of PCR product, add 1 μl of ExoSAP-IT enzyme (Shanghai Sinopharm Group, China) and incubate at 37°C for 15min, then treat at 80°C for 15min to inactivate the enzyme. After purification, the PCR product was diluted 4 times for use.

3.2 SNaPshot reaction and product purification

The SNaPshot reaction system was: 2 μl of SNaPshot reaction mixture (Multiplex Ready Reaction Mix); 0.5 μl of SNaPshot extension primers (final primer concentration was 0.2 μmol/L); 1.5 μl of the above purified and diluted PCR product; 1 μl of deionized water. The SNaPshot cycle parameters were 96°C for 10s, 50°C for 5s, and 60°C for 30s, for a total of 25 cycles. Take 5 μl SNaPshot reaction product, add 0.57 μl 10× buffer and 0.1 μl CIP (SNaPshot reaction product purification enzyme) and incubate at 37°C for 1 hour, then treat at 75°C for 15 minutes to inactivate the enzyme to achieve product purification effect, and store it at -20°C for later use.

3.3 Genetic analyzer electrophoresis and judgment

The electrophoresis mixing system includes: 9.25 μl Hi-Di formamide (formamide denaturant); 0.5 μl of the above-mentioned purified SNaPshot reaction product; 0.25 μl GeneScan 120LIZ size standard (electrophoresis molecular internal standard). After the system was mixed, it was denatured at 95°C for 5 minutes, and then immediately placed on ice for 2 minutes. Load by centrifugation. The specific judgment method is shown in Figure 3.

Table 1 Information of microsatellite and SNP marker primers used for fine mapping of ETEC F4ac receptor gene

Note: * indicates the markers used for genotyping of the F ₃ generation of the resource family.

3.4 Multi-point linkage analysis

The multi-point linkage analysis of 51 genetic marker loci and ETEC F4ac receptor gene loci was carried out using the public ALLEGRO software. The results of multi-point linkage analysis showed that the ETEC F4ac receptor gene locus was finely mapped to the 4.3cM region between SSC13UMNP997 and S0075 (Figure 4).

4. Breakpoint recombination analysis for fine positioning of ETEC F4ac receptor gene

In order to further narrow down the chromosomal region of the ETEC F4ac receptor gene, the applicant developed sequence tag sites (STS) and its SNP markers at a distance of 0.5 cM (see Table 2 for primers), and determined that white Duroc × Erhualian F ₂ (755 heads ) and F ₃ individuals (450 heads) genotypes at these SNP sites. Genotyping methods were performed using the SNaPshot method described above. According to the analysis of genetic recombination breakpoints at these sites in F ₂ and F ₃ resistant individuals, the occurrence of specific recombination patterns and changes in adhesion phenotypes were used to determine the breakpoint recombination of recipient genes to achieve fine mapping. In the F2 _- generation resistant individual with ear number 1487, the applicant found that a breakpoint recombination occurred at the S0283 marker in the UMNP997-S0075 region finely mapped by multi-point linkage analysis, thus determining the ETEC F4ac receptor gene between UMNP997 and Between S0283. Furthermore, breakpoint recombination was found at the UMNP358 marker in the UMNP997-S0283 region in the _F3- resistant individual with ear number 3314, so that the ETEC F4ac receptor gene was finally fine-located on the chromosome at 2.3cM between UMNP997 and S0283 area.

5. Anchoring of ETEC F4ac receptor gene

The applicant carried out high-density SNP search and identification in the target region of UMNP997-S02832.3cM and its two flanks by direct sequencing, and designed 34 pairs of primers in this region (see Table 2 for details), respectively in the distant population of Du Two phenotypic adherent and two resistant individuals were selected from Locke and Erhualian, and their DNA was used as a template for PCR amplification. The PCR amplification conditions were the same as above, and the PCR products were purified by QIAquick PCR Purification Kit (QIAGEN, Germany), and entrusted to Shanghai Huada Liuyi Company for sequencing. Using the SeqMan software in the public DNAStar software package to analyze and identify SNPs, a total of 104 SNP markers were identified. Select 70 evenly distributed and representative SNPs, and use MassARRAY Analyzer Compact for polymorphism in 292 individuals from 15 distant populations of Chinese and Western pig breeds that have been tested for adhesion phenotypes. Detection, time-of-flight mass spectrometry techniques are outlined below:

The PCR reaction system was 5 μl, which contained 8ng genome, 0.05mM dNTPs, 0.1μM primers, 4.0mM Mg ²⁺ , 1×PCR buffer and 0.5U Hotstar Taq DNA polymerase (Qiagen, Germany). The PCR amplification program was 94.0°C for 15 sec; 45 cycles (94.0°C for 20 sec, 56°C for 30 sec, 72.0°C for 1 min); 72.0°C for 3 min; 4.0°C for preservation. SAP enzyme (Qiagen, Germany) was purified to degrade dNTPs used for amplification to ensure that the extension reaction only extended one base. The SAP enzyme reaction system was 7 μl, which contained 5 μl of PCR product, 0.3 μl of SAP enzyme (1.7 U/μl), and 0.17 μl of 10× buffer. The procedure for SAP purification was 37.0°C for 40 minutes; 85.0°C for 5 minutes; 4.0°C for storage. The single-base extension iPLEX reaction system is 9 μl, which contains 7 μl of PCR product purified by SAP enzyme, 0.2 μl of iPLEX stop mixture, 0.2 μl of 10×iPLEX buffer, 0.94 μl of iPLEX extension primer, and 0.041 μl of iPLEX enzyme. The iPLEX reaction program was 94.0°C for 30 sec; 40 cycles (94.0°C for 5 sec, 5 cycles (52°C for 5 sec, 80.0°C for 5 sec)); 72.0°C for 3 min; 4.0°C for storage. Resin desalination purification reaction and sample loading: Take 6 mg of resin on a 384-hole resin scraper, cover evenly, scrape off excess resin, and place it for 15 minutes. Centrifuge the 384-well plate after the reaction at 4000rpm for 30s, add 16 μL of deionized water to each well, place it upside down on the resin plate, then buckle the resin plate on the 384-well plate by inversion, knock the resin into the 384-well plate, seal the film . Taking the long axis of the 384-orifice plate as the axis, turn the 384-orifice plate for 60 minutes, centrifuge at 3200 rpm for 5 minutes, and then set aside. The detection samples were transferred from the 384-well reaction plate to the MassARRAY SpectroCHIP chip covered with matrix. After the sample is transferred to the SpectroCHIP chip, it can be put into the mass spectrometer (SEQUENOM, USA) for detection, and each detection point only takes 3-5 seconds, and the analysis is fully automatic.

The test results were analyzed using public SHE software, and the analysis results are shown in Figure 5. It can be seen from the analysis result figure that the most significant SNP site associated with the adhesion phenotype, that is, M13A in the figure (the SNP at 46531bp in SEQ ID NO: 1), is located in the MUC13 gene. In addition, it has been reported that the ETEC F4ac receptor is a mucin-type sialoglycoprotein, and only three genes, MUC13, MUC4 and MUC20, express mucin glycoprotein in the applicant's finely mapped UMNP997-S0283 receptor gene region. The applicant's gene tissue expression experiments showed that the MUC4 gene was expressed in a very low abundance in the small intestine tissue, the MUC20 gene was expressed in a moderate and broad spectrum in various tissues, and the MUC13 gene was highly specific in the small intestine tissue (results not shown ). Combined with the SNP marker association analysis results of the above-mentioned distant populations, the MUC13 gene was identified as the ETECF4ac receptor target gene.

Table 2 Primer information for SNP identification in the target region of UMNP997-S0283

6. Identification of key mutation sites in ETEC F4ac receptor gene (MUC13 gene)

6.1 Isolation of new full-length cDNA sequence of MUC13 gene

Primers F1 and R1 (see Table 3) were designed according to the reported partial cDNA sequence of the porcine MUC13 gene (sequence number: EU046996) to obtain a cDNA fragment of 1148 bp. Using this sequence fragment, primer F2 was designed for 5' RACE to obtain a partial cDNA sequence. The conserved coding region of the carboxy-terminus of MUC13 protein was predicted by using the carboxy-terminal sequence information of human, bovine, dog, horse, cat and mouse by bioinformatics method. Then use the bovine conserved homologous region cDNA sequence to design primer F3 and perform 5' RACE again to obtain a new full-length cDNA sequence. The full-length cDNA sequence features are shown in the sequence table SEQ ID NO 1. The amino acid sequence encoded by the cDNA sequence of the MUC13 gene is shown as SEQ ID NO 3 in the sequence table.

6.2 Acquisition of full-length genome sequence of MUC13 gene

According to the obtained cDNA sequence of the MUC13 gene, the human-pig comparative genome method was used to design primer pairs M13-1-F/M13-1-R, M13-2-F/M13-2-R, M13 in combination with bioinformatics methods -3-F/M13-3-R, M13-4-F/M13-4-R, M13-5-F/M13-5-R, M13-6-F/M13-6-R (see primers for details table 3). The PCR reaction system included 25ng of porcine genomic DNA, 1.0mM MgCl ₂ , 100mM dNTP, 10pmol of forward and reverse primers, 2 units of DNA polymerase (Taq enzyme) and 1×PCR buffer (Shanghai Gaming Company, China). The amplification conditions were: 94°C for 4 minutes; 94°C for 30s, 55-65°C for 30s, and 72°C for 70s, a total of 32 cycles; finally extended at 72°C for 10 minutes. The amplified fragment was purified by QIAquick PCR Product Purification Kit and sequenced by Shanghai Huada Liuyi Company. Use the SeqMan software in the public DNAStar software package to analyze the sequencing results, and combine the existing cDNA sequences to obtain the full-length DNA sequence of the MUC13 gene. The full-length DNA sequence features are shown in the sequence table SEQ ID NO 2.

6.3 Identification of key SNP sites of MUC13 gene

Using the 6 pairs of primers isolated above for the complete gene sequence of the MUC13 gene, amplify the genomic DNA of Erhualian and Duroc resistant and susceptible individuals, the PCR reaction conditions are the same as above, and the PCR products are directly purified and sequenced, using SeqMan in the public DNAStar software package The software analyzed the sequencing results, and a total of 179 single nucleotide polymorphic sites (SNPs) were identified on the whole gene sequence of MUC13.

The 104 SNP sites identified on the full gene sequence of MUC13 were selected and used in a total of 292 individuals of 12 Chinese local pig breeds representing 6 ecological groups and 3 western imported pig breeds that had been tested for adhesion phenotype in vitro. Large-scale SNP genotyping by time-of-flight mass spectrometry. The correlation analysis was carried out between the SNP typing results and the adhesion phenotype of ETEC F4ac in vitro. The result is shown in Figure 6. Applicants found 5 SNPs namely SNP C53019T (M131009 in FIG. 6), SNP A46531G (M13A in FIG. 6), SNP A42784C (M13C4A in FIG. 6), SNP A32813G (M13C1B in FIG. 6 M13I-395) was used to determine the accuracy of susceptible/resistant individuals greater than 92%, the correlation P value was less than 1.18×10 ^-19 , and the accuracy of the most strongly correlated marker (C53019T) was greater than 97% (P= 1.59×10 ^-21 ). These five key SNP sites are markers in extreme linkage disequilibrium with the causative mutation on the MUC13 gene or are causal mutations themselves. According to their strong correlation with the adhesion phenotype, these 5 SNP markers can be well used in the production practice of genetic breeding for pig resistance to diarrhea disease, and the resistant individuals can be selected generation by generation through molecular marker selection to breed resistance to ETEC F4ac New lines of breeding pigs for diarrheal disease.

Table 3 MUC13 gene cloning primer information

The second object of the present invention is achieved in this way: the present invention utilizes 5 key SNP sites and flanking DNA sequence information of the MUC13 gene, and uses molecular biology techniques to identify individual genotypes to determine the individual's susceptibility to ETEC F4ac diarrhea. Susceptibility/resistance to breed improved pigs for diarrhea resistance traits.

Since 2003, the applicant has used the large-scale white Duroc × Erhualian resource family group constructed by the applicant to carry out _the research based on 183 _micro- Genome-wide linkage analysis of satellite loci, IBD analysis of high-density markers in the target region, and breakpoint recombination analysis, the ETEC F4ac receptor gene was finely mapped to the 2.3cM region adjacent to S0283 in the q41 region of chromosome 13 (SSC13q41). On the basis of fine mapping, a human-pig high-resolution comparative gene map of SSC13q41 including 20 functional loci was constructed. Further, through linkage disequilibrium correlation analysis of 292 individuals from 15 distant populations, the target receptor gene and the key mutation sites that can accurately determine susceptible and resistant individuals were finally identified, thus creating an efficient and accurate New breeding techniques for resistance to diarrheal disease in newborn and pre-weaned piglets.

The present invention identifies a MUC13 gene and its key marker sites that determine the susceptibility/resistance to diarrhea in newborn and pre-weaned piglets, detects the key mutation sites of the gene through modern molecular biology techniques, and uses marker-assisted selection (MAS) to select Favorable allele-type individual breeding can significantly improve the anti-diarrheal disease ability of weaned piglets in the population, and greatly reduce piglet mortality. The invention provides a new method for the disease-resistant breeding of pig ETEC F4ac diarrhea disease.

Description of drawings

Figure 1 The adhesion test results of ETEC F4ac and brush border, (a) is non-adhesive type, (b) is adhesive type;

Figure 2 Preliminary mapping results of genome-wide linkage analysis of ETEC F4ac receptor gene;

Figure 3 is the SNaPshot type judgment diagram of the C53019T locus, where a is the CC type; b is the CT type; c is the TT type;

Figure 4 shows the fine mapping results of multi-point linkage analysis of the target region of the ETEC F4ac receptor gene;

Figure 5 is a diagram showing the results of correlation analysis between the high-density SNP genotype in the target region of UMNP997-S0283 and the ETEC F4ac adhesion phenotype;

Figure 6 is a diagram showing the results of the correlation analysis between the high-density SNP genotype in the MUC13 gene and the ETEC F4ac adhesion phenotype.

Detailed ways

The present invention will be described in further detail below in conjunction with the embodiments and with reference to the accompanying drawings.

Select individuals from the core group of breeding pigs, use the SNaPshot method to detect the genotypes of 5 loci of MUC13 genes SNPA37617G, SNP A32813G, SNP A42784C, SNP A46531G, and SNP C53019T, and select individuals with favorable allelic types for breeding, and group selection Anti-diarrheal disease performance of post-improved populations in piglets.

Example:

144 purebred piglets including Duroc, Landrace and Large White pigs were used as experimental animals. These pigs were purchased from 5 large-scale commercial pig farms. Each pig breed selected at least 3 male lines, and each family selected 3-4 A 1-2 month old individual. After the 144 purebred piglets were slaughtered, the in vitro adhesion phenotype test of pig small intestinal epithelial cells ETEC F4ac was carried out, and the experimental methods and criteria were as described above.

The SNPA37617G, SNPA32813G, SNPA42784C, SNPA46531G, and SNP C53019T of the MUC13 gene were subjected to SNP genotyping in the above 144 purebred piglets using the above-mentioned SNaPshot method. The amplification primers are shown in Table 4. Statistical analysis was performed on the correlation between the genotype of each SNP site and the phenotype of ETEC F4ac adhered to pig intestinal brush border in vitro (see Table 5).

Table 4 is used to determine the primer information of 5 key SNP markers of MUC13 gene

The results showed that the genotypes of five loci, including SNP A37617G, SNP A32813G, SNP A42784C, SNP A46531G and SNP C53019T, were basically co-segregated with the adhesion phenotype. Taking the SNP site SNP C53019T as an example, the T allele is a susceptibility gene, and the C allele is a resistance gene; genotype TT and CT individuals are susceptible to ETEC F4ac infection, and CC individuals are susceptible to ETEC F4ac infection. Infestation is resistant. The CC type individual is the genotype individual required for the breeding and improvement of diarrhea-resistant pigs. In the core group of commercial breeding pigs, the subculture of CC individuals can significantly improve the anti-diarrheal disease traits of the population, reduce the mortality of piglets, and eventually breed new specialized strains that are resistant to diarrhea in newborn and weaned piglets.

Table 5 Correlation between the genotypes of the five MUC13 key SNPs in the experimental pig population and the adhesion phenotype of ETEC F4ac

sequence listing

                 

SEQ ID NO: 1

<110>Jiangxi Agricultural University

<120>The identification and application on a gene causing diarrhea in neonatal and young pigs

<130>

<160>2

<170>PatentIn version 3.2

<210>1

<211>6801

<212>cDNA

<213>Jiangxi Agricultural University

<220>

<221> CDS

<222>(41)..(5431)

<400>1

gaaaattttc tctctggatt acctggcacc aacatccaaa atgaaagtct tcattcactt 60

cactctgctt gccctccttt tcctaaaatc aggtaaggaa catttacaca tttttaattc 120

tttccgtcat atttcctctg ccattatata tacagtgttt tgtcttctga aactttgcct 180

ttcagcaagt atggctcaga acaagacagg aggcaccaca aagctgacat ccacagaaaa 240

caccagctcc ccgaacgcgg cggaaagcac cagcgctcct tccgctgcaa gaaacagtgc 300

tccgggcacg gcagcaagca ccagcgctcc gtctgcggca gcaagcacca gcgctccggc 360

cgcaggggca agcaccagcg ctccggccgc aggggcaagc accagcgctc cgtctgtggc 420

ggcaagcacc agcgctccgg ccgcaggggc aagcaccagc gctccttccg ctgcaagcac 480

gagcgcaccg gccgcagggg caagcaccag cgctccagcc gctgcggcaa gcaccagcgc 540

tccagccccc gcggcaagca ccagcgctgc cgcggcatca agcaccagcg ctccggccgt 600

ggcggcaagc accagcgctc cggcccccgc ggcaagcacc agcgctccct ccgctgcggc 660

aagcaccagt gctccggccg caggggcaag caccagcgct ccggaccccca cggcaagcac 720

cagcgctcag gccgcagagg caagcacaag cgctccgtcg gcggcggcaa gcaccagcgc 780

tccggccgca ggggcaagca ccagcgctcc gtctgtggcg gcaagcacca gcactccggc 840

cgcaggggca agcaccagcg gtccggccgc cacggcaagc accagcgctc cggccacagg 900

ggcaagcacc agcgctccgt ctgtggcggc aagcaccagc actccggccg caggggcaag 960

caccagcggt ccggccgccg cggcaagcac cagcgctccg gccgcagggg caagcaccag 1020

cgctccgtct gtggcggcaa gcaccagcac tccggccgca ggggcaagca ccagcggtcc 1080

ggccgccgcg gcaagcacca gcgctccggc cacaggggca agcaccagcg gtccggccgc 1140

ggcggcaagc accagcgctc cggctgtagg ggcaagcacc agcggtccag ccgcgtcagc 1200

aagcaccagc gctctgtctg aggcggaaag caccagctct ccagccgcag gggcaagcac 1260

cagcgctccg tctgtggcgg caagcaccag cgctccagcc gcggcggcaa gcaccagcgc 1320

tccttccgct gcaagcacca gcagtccgga ccccacggca agcaccagcg ctccggccgc 1380

aggggcaagc accagcgctc cctccgctgc ggcaagcacc agcgctccgg ccgcaggggc 1440

aagcaccagc gctccctccg ctgcggcaag caccagctct ccggccgcag gggcaagcac 1500

cagcgctcct tctgctgcaa gcaccagcgc tccgtctgtg gcggcaagca ccagcgctcc 1560

ggcccctgtg gcaagcacca gcgctccggc cccaggggca agcaccagcg ctccttctgc 1620

tgcaagcacc agcgctccgg ccacaggggc aagcaccagt gctccctccg ctgcggcaag 1680

caccagcgct ccgtccacgg cggcaagcac cagcgctccg gccgcagggg caagcaccag 1740

cgctccggcc cccgcggcaa gcaccagcgc tccggccccc gcggcaagca ccagcgctcc 1800

agcccccgcg gcaagcacca gcgctccggc tataggggca agcaccagcg ctccgtccgc 1860

tgcaagcacc agcgcaccgt ctgctgcaag caccagcgct ccggccgcag gggcaagcac 1920

cagcgctccg tctgtggcgg caagcaccag cgctccggcc cctgcggcaa gcaccagcgc 1980

tccggccgca ggggcaagca ccagcgctcc ttccgctgca agcaccagcg ctccagccgc 2040

aggggcaagc accagcgctc cggccgcggc ggcaagcacc agcactccag accccacggc 2100

aagcaccagc gctccggccg caggggcaag caccagcgct ccggcccctg cggcaagccc 2160

cagcgctccg gccgcaggga caagcaccag cggtccagcc cccgcggcaa gcaccagcgc 2220

tccggccgcc gcggcaagca ccagccctct ggaccccacg gcaagcacca gcactccggc 2280

cgcaggggca agcaccagcg ctccatcggc agcggcaagc accagcgctc cttctgctgc 2340

aagcaccagc gctccttctg ctgcaagcac cagcgctccg gccgcagggg caagcaccag 2400

cgctccggcc gtggaggcaa gcaccagcgc tccggccccc gcggcaagca ccagcgctcc 2460

ggctgcaggg gcaagcacca gcgctccgtc cgctgcaagc accagcgctc cggccgcagg 2520

ggcaagcacc agcgctccgg ccgcggcagc aagcaccagc gctccggccc ccacggcaag 2580

caccagcgct cccgccgcgg tggcaagcac cagcgctgcg gcccccgcgg caaccaccag 2640

cgctctggcc gcggtggcaa gcaccaaccc tccggccgcc ggggcaagca ccagctctcc 2700

ggccgcaggg gcaagcacca gcgctccgtc tgtggcggca agcaccagcg ctccggcccc 2760

tgtggcaagc accagcgctc cggccgcagg ggcaagcacc agcgctcctt ccgctgcaag 2820

caccagcgct ccggccgcag gggcaagcac cagcgctccg gcccccgggg caagcaccag 2880

cgctccggcc cccgcggcaa gcaccagcgc tccttccgct gcaagcacca gcgctccggc 2940

cataggggca agcaccagcg gtccagcccc cgcggcaagc accagcgctc cggccgccgc 3000

ggcaagcacc agcgctccgg accccacggc aagcaccagc actccggccg caggggcaag 3060

caccagcgct ccgtcggcag cggcaagcac cagcgctccg gcccccgcgg caagcaccag 3120

cgctccttct gctgcaagca ccagcgctct ggaccccacg gcaagcacca gcgctccgtc 3180

cgctgcaagc accagcgctc cggccgcagg ggcaagcacc agcgctccgg cccccgcggc 3240

aagcaccagc actccggctg caggggcaag caccagcgct ccttctgctg caagcaccag 3300

cgctccggcc gcaggggcaa gcaccagcgc tccggccccc acggcaagca ccagcgctcc 3360

cgccgcggtg gcaagcacca gcgctgcggc ccccacggca agcaccagcg ctccggccgc 3420

ggcagcaagc accagcactc tggcccccac ggcaagcacc agcgctccgg ctcccgcggc 3480

aagcaccagt gctccgtccg cggcaagcac aagcgctccg gccgcggcgg caagcaccag 3540

cgctccggcc cccgcggcaa gcaccagcgc tccggctggg gcggcaagca ccagcgctcc 3600

ggctgcaggg gcaagcacca gcgctccatc cgctgcaagc accagcgctc cggccgcggc 3660

ggcaagcacc agcgctccgg ccgcaggggc aagcaccagc gctccggccg ccgcggcaag 3720

caccagcgct ccggcccccg ctgcaagcac cagcgctccg gcggcggtgg caagcaccag 3780

cgctccggcc ccaggggcaa gcaccagcgc tccatccgtg gcaagcacca gcgctccggc 3840

ccccgctgca agcaccagct ctccggccgc aggggcaagc accagcgctc catccgcggc 3900

aagcaccagc gctccggtcc ccgctgcaag caccagctct ccggccgcag gggcaagcac 3960

cagcgctccg gcccccgctg caagcaccag ctctccggct gcaggggcaa gcaccagcgc 4020

tccggccgcg ggggcaagca ccagcgctcc ggccgcaggg gcaagcacca gcgctccagc 4080

ccccgctgca agcaccagct ctccggccgc aggggcaagc accagcgctc cggcccccgc 4140

tgcaagcacc agcgctgcgg ccgcgggggc aagcaccagc gctccatccg cggaaagcac 4200

cagcgctccg gctgcagggg caagcaccag cgctccggct gcaggggcaa gcaccagtgc 4260

tccggccgcg ggggcaagca ccagcgctcc atccgcggca agcaccagcg ctccggccgc 4320

cgcagcaagc accactcctg cctctgcagc aaatgcttct tctgctattt cgagtacacc 4380

tattgcctcc acacccacct caggttccag gaacccttgc cagcaagatt cctgtaaggg 4440

tggttcttca tgtgttagtc tgaataatac gtttttttgc ctgtgttcag aagggtatta 4500

ctataactct tccacctgca ataaaggaaa gattttccct ggtaaaatta cggtcaaaat 4560

atcagaaaca actgatctag aacatgaaaa ttctgtgacc tatcaaaatt tataccttca 4620

agttacaaat ttttttaatg acaatttag caattctgag tacggacaga ccgtaattct 4680

taaagtaagc acaactcctt cagcaagata tgaaatgcgt gccagcaacc agcttgttga 4740

ggtagaagta gtaaacattt ttagagaaac tacaacagaa aatgagagtt ccatatttaa 4800

tgccattgac caagcaatta aaagcaaccc aagtgacttt acagaatata tccagcaaga 4860

tcggtgtgat tattatggat gtaagaggca aaatgagcaa gatgagtgcc ccagtggttt 4920

actgtgtgag tgcaaagagg ggctggagag accaaaccca cagatccctc tgtgtgttgc 4980

tttgggtcca gcatgctctg atacctgcaa tgcagagaac aataggcaat gcctcgtgag 5040

gagcgctggg aacgccgaat gcttgtgcct gcctggctac aaggaggata aaaacaagat 5100

ctgccaagcg tgtccatttg gctacagtgg agttggctgt gaaaacccat atcagctgat 5160

cctcaccatc gtgggcacca tcgctggcat cctcattctc agcctggtga ttgcattcgt 5220

cctctcatta agatcaaaca acaaaaacaa gaatgtggaa gaacagaact tgattgagaa 5280

ygactttcaa aacctgaaac tgcaggagac tggctttagc aacctagag caggaaacct 5340

ctttcctaag atcaaggtgg aatttccag aggtggtcag ccacagaatc cctatataac 5400

ccmgggtagt ataccccgcc ctgactatta gaatggtaaa aatttggaac catccatggc 5460

cgtcaaccca gcacatgaga tttttgtgtg agggtttgat agactgatgg agaaaagaga 5520

tctggccttt agtatttttc ctccatccaa catctggtct ctctgaatgg aagctgtaac 5580

tgtttgcatt gaattcagcg tgtttgaatc taagtaccta tgaagttaaa tgcacacata 5640

aatgctatta gcacttgtaa gatagttgat ttgctccagt acagtggtta ggtgtttgtt 5700

tgttttttcc tggtgggaca acaataacca tttccaaact agagggaagc tccccaggat 5760

tgttcattcc tggaagagtc taagtgttgc tttgagttga tagcatgcag ataagtgacc 5820

taattctcgg aaattcttac acatcagctg tggaattcca gtagacggga ggagacacca 5880

tccttaaagg tactcagtgg accctgaaga ttcattctct agcaagttaa aataaggatg 5940

ctacttctgg tcctcaaatg tgggaatttc attaccctta aactgcttgc gacctggctt 6000

cacaccaaga ggacagaaaa caagacatgt ctcccagctt tcaggaagga gttcctggga 6060

atccaggacc tttgagatgt taattaaaag atgtggactt gggagttccc gtcgtggcgc 6120

agtggttaac gaatccgact aggaaccatg aggttgcggg ttcggtccct gcccttgctc 6180

agtgggttaa tgatccggcg ttgccgtgag ctgtggtgta ggttgcagac gcggctcgga 6240

tcctgcgttg ctgtggatct ggcgtaggcc ggaggctaca gctccgattc gacccctagc 6300

atgggaacct ccatatgccg cgggagtggc ccaagaaata gcaaaaaaaa aaaaaaaaaa 6360

aaaaaaaaaa aaagatgtgg acttgagatg tcttccatga ggatacattc tagagtcttt 6420

gaaatgctgg tgggtgagac caagagtagg agcaacagag gcagaaggaa gctgagctgg 6480

gaggaagaag gtacccaaga cccaccttca tcctcagatt cccctcccca ataaacaggt 6540

gggcactttc tctaatgcat gccctacta tgactcaaga cctatgaaag gatgtgaaga 6600

taaaaaccct ggagcaagag tttggagtca ctttgaaatt gtagacattc tgtctggctg 6660

atggtggaga agtgatgcag attccagagg tacttcacca aagtctcagc cttccctggg 6720

gacagaggct tagcagtggc caccaacaca cacacgcaca tacacagcca agaatggggc 6780

agccacctgc cgcacaagct t 6801

SEQ ID NO2

<210>2

<211>56728

<212>DNA

<213>Jiangxi Agricultural University

<220>

<221> exon

<222>(20240)..(20373)

<220>

<221> exon

<222>(22970)..(27238)

<220>

<221> exon

<222>(32212)..(32334)

<220>

<221> mutation=A/G

<222>(32813)..(32813)

<220>

<221> exon

<222>(33499)..(33605)

<220>

<221> mutation=A/G

<222>(37617)..(37617)

<223>Y=C/T

<220>

<221> exon

<222>(39137)..(39192)

<220>

<221> exon

<222>(42484)..(42647)

<220>

<221>mutation=A/C

<222>(42784)..(42784)

<220>

<221> exon

<222>(46287)..(46412)

<220>

<221> mutation=A/G

<222>(46531)..(46531)

<220>

<221> exon

<222>(46763)..(46892)

<220>

<221> exon

<222>(47846)..(47883)

<220>

<221> exon

<222>(50196)..(50280)

<220>

<221> exon

<222>(52971)..(53169)

<220>

<221>mutation=C/T

<222>(53019)..(53019)

<223>Y=C/T

<220>

<221>5′UTR

<222>(55361)..(56728)

<400>2

accgcggcca tagtcgatcg ttctgctgtg catgatggga attcccataa tggttgtaat 60

tttgatgtgt ctgtgggagg aggtgaattc agagttcttc tattccacca tcttggccac 120

acccaatagc agacatta tgagccatag gaaatgaatt ctatgtccca tttctagtct 180

ctgaggaaat gccaatgctt caacattttc tttcttaaat caccaatgca tatgcatcat 240

gacaaaaaaa aaaagaaggt aaaaatggac ttcaagtgcc attcttttaa atcccagtat 300

ggattatttt agactttgaa gtatggatta tttttttatt caacttgatg gcaaagcatt 360

atctatcatg cagtgacact gttggtgggc taaaataata catttatgtta atttaccaga 420

ctaagcacgt atcacagtat ttccaactca cagaaaagca accatcaggg aaaatagaaa 480

gttttaagcg gaatatctca ttgtagcaga atttccccgt aaaattaaat aatgaaaatg 540

aggctgcaat caaagcaagt tttcaaatgg ctcatttgtt agccaagcca ggaaagccat 600

ttagggatga gttaattaaa ctatttgatt gcagtagtca aagaaatgtg tctagaggaa 660

agaaacttgc ttaagactgt ttcagcaaga acagttactc agaatgttga gaacagtagg 720

aacagaatca atggtcaatt aataaacaag gtacataatt ttgaatggtt tccttggctc 780

atgatgagtt gtaggtgtt accaatagca ctcaattttt gtttattcca ggattcaatg 840

acaagtttga agtgactgaa taattatcct ctgtgaacag attctgtgga acaactatag 900

gaaagaatat tttcaaaata aaattgtata ttttaaatgg ataaattata agtgaattaa 960

atctcaataa ggctcttttg taaaaaagaa tgatttcaaa gaagttgaga aaacactatt 1020

ttcagtataa actgaagtag aatctgccaa gatacaactg aagatagtag aaatatgtgg 1080

ggagtagaaa aagttatagt tggattcatt tatagtgtgt gaaaatgtta gacatttaaa 1140

tcctctggtt attcattaca ttattatga ataagtattt tgtgggaaat atttaaatac 1200

gtcatatgct atcgagccag cactgtaagg gtgaacttca ttcattgtca gtggacttaa 1260

ccgtcattgg ttccatgaaa tttttttgtt ttgttttgtt ttgttttgtt tttagggcca 1320

cacccatgac atatgaaggt tcccaggcta gagtctaat ctgagttaac agctgctggc 1380

cacagccaca gccatagcaa ggcaggatcc aagccgcgtc tgcagcctac accacagctc 1440

acagcaactc cagatcctta accactgag caaggccagg gatcaaaacc tcaacctcat 1500

ggttcctagt tagattcatt tccattgtgc ctcgatagga actcccaggt ttggtgaatt 1560

tttgtgggat atagaagcta ttatcctgac ttgccctgct ataaagcagt tggatggctt 1620

agcagtaaaa aagttttt gcaagttttt gagcatagggg ttaggattaa aatctttctg 1680

agtagaacca gtctaaacca cttgcattaa acgggagtgg tttttgaaga tgtcttttgc 1740

tgcagacttg ataatgcttc tcaatgaatt caactcacaa ttactcaagg caaaacagct 1800

ctacatgcaa attttatgct atggtaaagt cattttaaca acaatttaat aatgttgtgt 1860

aattcacaag taatgtcaag ctactttcta taattcccat gtagtcagaa ttttaaatga 1920

gaagatcttc cttcccacat gaatttcagc agacatattt tctggactta aagtacaatt 1980

ccagcagtgt tttgaaaact tcagtacaag agcatagaaa atttctgtat ttcaaaatcc 2040

atactatgca attgaggagc agttacctaa cattcaatcg ggaatgatca atttgcaatg 2100

taatggcgtg tcaaaaggca actatcaata gaagaatcta atagaatgct gtaactgcct 2160

ttcaagcaat gagtatactt aactaaagct acatgttcat ggactgatac tagaatttgg 2220

cagaactacc tatgtgtatg aaaagatatc ttcaaaaatt aaatataggg agttcccatc 2280

atggctcagt ggttaacgaa cctaactggc atccatgagg accccaggttc aatccctggc 2340

cctgctcagt gggttaagaa tctggcattg cccagagctg tggagtaggt cacagacgtg 2400

gcttggatcc cgagttgcta tggccctggc gtaggctggc agctgtagct ccggttggac 2460

ccctagcctg ggaacctcca tgtgccgcat agccctaaaa aagagagaaa gacaaaataa 2520

aaaaaaaaga aagaaatata taaatatcat tataaatctg tattaacagt tggacatttg 2580

tgatacattt tgatgatata gggaatctga ctctgaacct caattaagca aaatattatc 2640

gctccccaaa taattccatt ctctatcaat aaaccagtat caccaaatac tgtgctcaat 2700

actattatat tttaaatttc atcagtaaat ttttttggaa atttgtttcc tctttagtat 2760

acataagtgt ctacattata tcttcaattt tccctcttgg ctcacaaaac ttgaaatatt 2820

taatatctag gagttccctt atggcatggc aggttaagaa tctggcattg tcactgcagc 2880

agcttggctt gctgctgtgt ggaccgagtt tgatccctgg cctaggaact tccacatacc 2940

acaggcacag ccaaaaataa tttactgtct agccctttat taaaaaagtt tgctgattcc 3000

aatctatgtc ccagtcctac ctatgcttca gatttctaac cttagcattc atattataac 3060

agtaaatctt cttcatggtt ttgagttaat aagtgcaaag tgtatagcat aatggcaaag 3120

ggtaggtgct ttttaagtat gtttttatta atattaagaa ctatattgct atagctcttt 3180

tgaactatac agagttgtgg tcattacaaa agatgatctg taggcagagg gaggcctctg 3240

gtgaataaag atttccttta agaaaatctt ctttatcatt tttttttt atgttttagt 3300

ggattatagt tcatttacag tactgtgaag acagtcctct ttaaaactgc cctggggtgg 3360

gggaggcttc cacttaacgc atcccagctt gctttcttaa aattttgttt ttttggtagc 3420

agctttactg agatacaatt cacataccat aaaattcaca cattaaaatt cactgcatca 3480

tcacaaccaa ttttagatta tcttatttta gccatcatct ccccatcctc ccacctcccc 3540

ccagccatag gcaacaccta atcctctttc tttctctgta gatttgcctg ttctggaatt 3600

taatgtaagt ggaattatac atgtggcctt ttatgactgg ttatttcat ttagcatatt 3660

actttttttt tttttttttt gtctttttgc catttctggg gccgctctca cagcatatgg 3720

aggttccccg actaggagtc taatcggagc tgtagccact ggcctacgcc agagccacag 3780

caacgcggga tccgagcctc atctgcaacc tacaccacag ctcatggcaa tgccggatcc 3840

ttaacccact gagcaaggcc agggatgaa cctgcaacct catgattcct agtcggattt 3900

gttaaccact gcgccacgac gggaactcct agcatattac tttcaagcct tctccacatt 3960

gtggcatgta tcagtacttc attccttttt tggtggagta gtgttccatt gtgtgagtgt 4020

accccacttt gtttatccat tcatcagttg atggacattg ggatatttct gtgtttgttc 4080

tcttgtgaat aatgaacatt cacatacaaa tattatgtg aacaatgttt tcacttcttt 4140

taggaatata cctagaagtg aaattgctgg gtaaaatggg aactctatgt ctaacttttt 4200

gagaaactgc cagatttctg tccaaagtag ctgtgtcttt gttttttttg ttgttgtttg 4260

ttttttgttt gtttgtttgt ttttggtctt tttagggccg cacctgcagc atgtggaagt 4320

tcccaggctc ggggttaaat ccaagctaca gctgccaacc tgtgccacag ccacagcaac 4380

gccagatctg agctgggcct gcaacctaca ccctagctca cggcaacgct ggatccttaa 4440

cccactgagc ggagccaggg atcgaaccca catcctcatg gataccagtt gggttcctaa 4500

ccactgagcc tcaacaggaa ctccagctgt gccatgtttg catccttacc agcagtgtat 4560

gaggagtcca gttcccccac gtcgccacca acacttgaaa ttcaattatc tggctctttg 4620

attgtaggca tcctagtgat gataaagtga taaccagctc atttttaatc ctctgggatg 4680

gggtattgggg gaagctattc tttttaacac tccacacagg gtctacgggg gaacagccct 4740

ttggaaactt agccctgcaa gatcatcagg gtatttccac tgaaactgaa gtttttctca 4800

gagggattgt gccccattag taactcgtcg tcgatttgta acaacccaga tatttgcacc 4860

accctcttag tagatattat tgggagatga gcacatattt ttgagggact acttacagtt 4920

acaaggcaga ttattacaag gcatctcgac tccacaggca ccaggacaag atcccttcta 4980

gagagaagtt ctgtagacac gactgcccag ccccacctcc caccacctat ttcccatccc 5040

tgtgatttgc agacaccaag gacccttctg gggagagcaa ccagagctgc tgggaaagtt 5100

tccataaaga aaggaagagt ttctctgggg aggcagacac cacagcacag caagagctca 5160

gaagggaagt tggttcctgg tcctgccctg tgtggtggct gcttcccagg gtggaagaga 5220

aattgaactc tttgcaccca aagcaagtca gccagtcaag ggctgtttac tgggtgcctg 5280

catatgctga cgtcccttgg gtgttgcact tactgggtgc accttcagag agtgtttcaa 5340

agtcaggtct tcccattgca tgctcccact ccaccctgag ggtgtcttgc atagcacttt 5400

gcacagttat ggcagaaatg ctaatttcag ttcattgctt ggtacccaga gcctggaaca 5460

gcatctggca tgtgcacata ctctaaagta tttggtgaat gaaggaaaaa aatctaaacc 5520

tgacactggc tagtggaatc caagcacagt agtacctcag catatattat tttgtagttt 5580

atttgtactt agtaaatata gcgttgttct cctgttgtta ggtattccct gatggcccag 5640

tgcttaagaa tatggtgttg tcactgctgt gacttaggtc accattgtgg cgctggtttg 5700

atccctggcc taggaacttc tgtaggcctc aggcgtgcag ctgaaaaaaa agatgcagta 5760

ttgttgaaaa tttcatttgg catagtgact atgtaacatt cttctcatgt acagtgaggt 5820

accacttcac actagtcaga atggccatta taacaagtca acgaataaca aatgctggag 5880

agagtgtgga gagaagggac ccctcctttg ctgttggtgg gaatgtaaat tggtacaacc 5940

actatggaaa actaaacatg gaagtaccat atgatccagc aatcccattc ctgagcatat 6000

atccagacaa acatttgttc aaaaaaatac atgaaccctt acgttcatca catcactatt 6060

cacaataggc aagacatgga aacaacttaa atgcccattg atagatgaat ggatcaagaa 6120

gatgtggtac atatacacaa tggaatacta ctcagccatt aaaaaggaca aaataatgtc 6180

atttgcagca acatggccag aactagagat tctcataacta agtgaggtaa gtcagaaaga 6240

ggaagatatg acatcatttg tatctggaat ccaaaatatg gcacaaatgg tcctatctac 6300

aaaacagaag ttcccgtcat ggctcactgg ttaatgaatc ttacgaggaa ccacgaggtt 6360

gtgggttcaa tccctggcct tgctcagtgg gttaaggatc cagcgttgtc gtgagctgtg 6420

gtgtaggtca cagacatggc tctgatcttg tgttgctgtg gctctggtgt aggccagtgg 6480

ctacagctcc gattcgaccc ctagcccagg aacctccata tgccacgggt gcagccctgg 6540

aaaagacaga aagacaaaaa aaaaaaaaaa ccaaaaaaca gaaacagatc atggacatgg 6600

agagcagact tgtgtttgct ggggcaggga gggaaggag tgggatggat ggagagtttg 6660

gggttggaag atgcaaaata ttacatatgg aatggataag caatggggtc ctactgtaca 6720

gcatagggaa ccaagtctaa tgtcttgggt tagaacatga tggaagatat tattaaaaa 6780

aaagaatgta catgtatgta tgactgggtc actttgctgt acaacagaaa ttgaaggaac 6840

attgtaaatc aactacactt taataaaaaa aattttttta aatattctcc tcacaaactg 6900

accaaatttg tcctgaccaa gctctggcta ttggatattt acaattgttt aaaatttttt 6960

cctatgaaaa aataatagtg ggcttgggct gtgggatgga aatcctgtga aatcagattg 7020

ttatgatcat tatacaacta cagatgtgat aaattcattt gagtaataaa aaaaatgaaa 7080

aaaaaatagt gggagttcct gtcgtggtgc agcggaaatg aatctgacta ggaaccagga 7140

gattgggcgg gttcgatcct tggcctccct cagtggattc aggatctggc gttgccatga 7200

gctgtgatgt aggttgcaga cgcgcctccg atctggcatt gctgtggctg tggcctagtc 7260

tggcagctac agctccgatg agccccctag cctgggaacc tcatatgctg tgggtgcggc 7320

cctaaaagga caaaagacca aaaaaaaaaa aaaaatagtg atacatcctt gtagaaatgt 7380

ctttgagtac atatctgact gcttccttac tataattccc ggaatggaaa tcctgggtcg 7440

aggaacatgc ccgttgtcaa aggtccatga tgcctatttg ccagcatccc ttctcccaag 7500

ggctgtggat ggggcctgtt tccccaacac cattgacctc agcaagcatc gtcattttct 7560

tcatgctttc agtaaaattg ccccatagtc agaatatctg attcctgata ctacacagcc 7620

tctcagggga ttgcctgtga atgcaggccc aagttttatg tcaaatttgc cataaagaaa 7680

atagttcctg agaaaaccag atcaaggaaa ccagatttcc ttcttggaaa agccatttta 7740

tagatgacta ataataacca ctaccaaaaa gaatgtctgg ttaaagattc ctggaaaagt 7800

tgcaggttga gtgtgcgcct tgaacgtcaa gttaagtgtc atacactttc cccagactgc 7860

cagtcagctg ggggttgggg gtgacttccc aggaagaggt gcttatctga aaataattta 7920

gttgctagaa agtaactcag ggtgcggctc cagcaggctt gattcaattt tctcttttac 7980

tacattgatg tgacgtttgt catgttactg atatatatat ttttaattta attcacctgt 8040

ttgcttcccc tgtccccaca cttttaaagg gcctcttccc taacatccag ccaaccatgt 8100

gtcctagcag attttccata caaggttctt gacacaccgt gtatctctgc tcttctgtaa 8160

tctttttctt gaaattgact tattttttaa agacctcaat gaatttattt ttaaaaaatt 8220

tattggagga cgttcccatt gtggcacaga tgaaatgaat ctgactagga accatgaggt 8280

ttgtgggttc gatccctggc ctcattcagt gggttaagga tccagctttg ccatgagctg 8340

tggtgtagtt cgcagacgca gcttggatcc tgcgtggctg tggctctggc gtaggcttgc 8400

cagctacagc tctgatttga cccttagcct gggaacctct gcatgctgag ggtgcggcag 8460

taaagagaga aaaaaaaaca aaaaaaattt ttggagtata gttgactcac aatgttgcat 8520

tagtttcagg tgtacagcaa agtgattcag ttacatgtat acatgtatcc attctttctc 8580

agattctttt cccatgagct tattagagaa tattgagtag agttccctgt gctatgcagt 8640

aggcccttgc tgattatcta ttttagatat attagtgtgt acatgttaaa cctgaactcc 8700

taatttatcc ctcccccacc atgatttccc ttttggtaac tgtaaatttg attttgaaag 8760

ttgtgagttt gtttatgctt tataaataag ctatcttgta tcacttttga ttagattcca 8820

catataagtg ttatcatatg atatttacct ttatcttgct gacttcactt agtatgataa 8880

tctctaggtc tatccacgtt gctgcaaatg gcactatttc gttctttttt atggctcagt 8940

aatattccat cgtatacatg aaccacatct tctttatcca cttttccatc aattccacgt 9000

cttggctact gtaagtagtg gctactgtaa atagcagtga aattgggat gccagtatct 9060

ttttgaatta tggttttctc tgcatagatg cccaggagtg ggattgctgg attatatggt 9120

acttctatat ttagtttttt aaggaaactc catactgttt ttcataacag ttgcaccaat 9180

gtacattgcc accaacagtg taggaggatt cccttttaaa tgaatttgct tgtaaagtta 9240

actttacatc attacctcaa attaaaaact agtttacttt tccatgaata ggaaggcacc 9300

ataaaacaaa tacaataaaa ttctaggtgg atctctgtga ttaccaaagt ctctttgagc 9360

agttttcttc ctggtagtgg gaaataggca ggcattcgag aacagttaaa gatataccag 9420

cagtaaatga gtggtataat cagaatgatt gacataaatt tgaaaatgca attaccttcc 9480

ccgtcaatgt gaaccattgt tctctaacgc tatgtccaca ggccacttaa aattgtcttg 9540

tgtgtcacaa gcacagtagc tgttatacct gagtgtgcat cagagtttcc tggagtctcg 9600

ttaaaacaca gatcactgat taagtctggg ataaggcttg aaatgtgcat ttctaaaaag 9660

tgcccaagcc agagttcccg ttgtgactca gtgggttaag aactgactag tacgtggagt 9720

tcctgttgtg gcacagtggt taacgaatct gactaggaac catgaggttg cgggttcggt 9780

ccctgccctt gctcagtggg ttaatgatcc ggcgttgccg tgagttatgg tgtaggtggc 9840

agacaaggct cagatcccat gttgctgggc tgtagtgtaa gccagcagct gcagctccag 9900

ttccaccccct agcctagaaa cttccataga ccacaaatgt ggacctaaaa agaaaaaaaa 9960

ataaaataaa aaattaaggg cattcctgtt gtggttacga acctcactag tatctatgag 10020

gttgtgggtt cgatccctgg cctcactcag tgggttaagg atccagcatt gctgtgagct 10080

gtggtgtagg tcacagatac agcttggatc ctgcatttgc tggtctgcca gcagccacag 10140

ctctgattcg acccttagcc tgggaacttc catattccgc aggtgcggcc ctgaaaagca 10200

aataataata ataataataa attaaaataa aaacataagt gcctaagcca tgctggcact 10260

gctagttcag tgcacacact ccaagaggca gaaccctcat ttcatcacct ctcactttaa 10320

gaaaaactgt ctggggaagg cttacagatg caaagcaaaa acaactgcag gtacaaacag 10380

gtacaaacct cctcccatcc cctgtgggca attttccttt caaagagcca caagcccagg 10440

ctaggcacta ccctccctga aagcagtgct gtaatctcta gacccaccca cttcctctag 10500

aggaagcctg aattaaaatg tggagccggg agagcctt ctgatcttag taaaacctcc 10560

ccaccatagg tgagcagagg aggaagtaga tgagaaaccc aactactttc ttggaaaagc 10620

ccctccatat aatttttccc agagctgaat ttctagaaga tcaaaaatca aatggaatta 10680

agaaatccac agaaattcct ttttttcttt ttctttttag ggccgcatcc acagcatatg 10740

gaggttccca ggctaggggt ccaatcagag ctatagcagc cagcctacac cccagctcac 10800

agcaatgcca gatccttaac ccactgagcg aggccaggag tcgaacctgc atcctcatgg 10860

atgctagtca ggttcattaa ctgccaagcc acgatgggaa cttcaaaaaa gcattttttg 10920

agaactgact aaactcagtg aggtgagtag atgagtgatt gattgtggag tttcacacgc 10980

caagatgcaa acacctatgc ctcccccttt tgaaggccga ggtattggga atgcagctcc 11040

agtgagtgaa ggtccttgtg tcagtattag aaccttcctc tgagcccttg atatcactgg 11100

tgtgtgagaa gagaaacatc tcaggaagac agctcaacca cgatcctgcc tcccagtaag 11160

tcaaagcaca ttaatttgtc catgtttcct gctagagctt ttctttatag aggcacattt 11220

ttcatgtaat tagatatata atgataacct gctttttttt ttttttttgc tttttagggc 11280

tgcacctgca gcatatggaa gttctccggc taggggtcga atcggagcta cggtttccag 11340

cttacaccac agccatggca atgctgggat tgaacctgca tcctcatgga tactagtcag 11400

attcatttct gctgtgccac aatgggaact cccaagactc ttgcatttaa aaaaaatttt 11460

attgaaatag atgaagtatt gttgattac aatattgtgt taattcctgg tgtacaaagt 11520

gattcggttt ctcatctata ttcttttcca ttatagtttg ttacaggata ttgcatgtaa 11580

caggatatag tttgttgcag gatattgtgt tatacagtag gaccttctcg tttatctatt 11640

ttatatattg tagggtttt tttttttgtt ttttttttttttgctatttc ttgggccgct 11700

tccgcggcat atggaggttc ccaggctagg ggtccaatcg gagctgtagc caccggctta 11760

cgccagagcc acagcaacgc gggatccgag ccgcatctgc aacctacacc gcagctcaca 11820

gcaacgccgg atcgttaacc cactgagcaa gcgcagggac cgaacccgca acctcatggt 11880

tcctagtcgg attcgttaac cactgcgcca cgacaggaac tcctatattg tagtttgtat 11940

ctgctaatac caaaccccta atttattcct cccccacccc cttttccctt tggtaaccat 12000

caatttgttc tctatgtctg tgagtttgtt tctgtttcct aagtaagttc atttgtatca 12060

tattttagat tccatgtata aatgatattg tacggtattt gtctttttct ttttgagggg 12120

tgggccacac ccatggcatg cagaagttcc ctggccaggg gtcgaacctg tgccacagca 12180

gcaacccgag ccacagcagt gacaatactg gatccttaac cttctgcacc acaagggaac 12240

tcctgtcttt atctttccaa cttacttcac ttactataat catttctaat tgcatctatg 12300

tttctaggaa tggcattgtt tcattctttt ttatggctga gtagtattcc attgtataat 12360

attatcacat cttctttatc cattcctctg ttgatgaaca tttaggttgc ttccatgtct 12420

tgtatattgt acataatgct gcaatgaatt ttgagatgca ggtatctttt agaactaaag 12480

tttttctcca gatatatgcc aagagtggga tttgttggat catatggcaa ctatattttg 12540

gggtttttgt tttgtcttgt tttttttctc ttcttagggc cacgcctgca gtatatggaa 12600

gttcccaggc tagggtcaa atgggagctg cagcagctgg cctactccac agccactgca 12660

acttaggatc tgagctgcat ctacaaatat atgctgcagc ttgtggcaac atgagagact 12720

taacccaccg agcaaggcca gggatcgaac ctgcatcctc atggatacca gtcaggttct 12780

taacctgctg agccacaaca ggaactccct gtattttttt tttttttaag gcacctccat 12840

gctgtcttcg atagtggctg caccccatta cattcccaaa caatggtgta gagaggagaa 12900

ccctgccttt tccacacttt gtcacagtag aataagcact ctcttttt gctgtaccta 12960

ggcttccctt gtgcgatgct gtcatatatt tcctcaagga gagtcaccac atttattcct 13020

ctgtccttga atagtagcag ttgtttttaa ttgttttttg gtaataatgt aaaagtgcaa 13080

caaaagttat tgtacatata attttccctt catttggaaa tatttcctta gggtaaagtc 13140

ctagaaggga atttctcagt ctaaagataa gaacattttt atggccctta atatctctcg 13200

ctgaattgcc ttttgaaggg gatgtagcat ttgcaagtgt gtgaaattat ctgaaccccc 13260

accagaattg aatattatcc atgtgaacat ttttaccaaa tgtaaaatat tacttcattt 13320

gtgttctagt cagtagcatg tcttttttgt ttgtttgctt gtttgttttt taatcttttt 13380

agggccacac ctgcagcata tggaaattcc caggctaggg gttgaatcgg agctgcagct 13440

gccagtctac accacagcca cagcaacgtg ggatccaagc tctgtctgcg acctacaccc 13500

cagctcatgg caacaccgga tccttaaccc actgagtgaa gctggggatc taacccacat 13560

cctcatggat gctagtgggg ttcgttacca ctgagacacg aggggaactc ccagtatcat 13620

gtctttaaaa tcagtgaaaa gtcaagcatt ttcccatata tctttactaa ttttattttc 13680

tctttgtgaa taatttatca tgacaatcga cttttggaaa aggaattgtc caaatgcata 13740

gtgagtctga agaccaggac tatatgttat ttactttgca ctccctgctc cctgtatcta 13800

caacaaatgg gatgacgttc aactattctt ttctttattc cataaatact tactgggtgc 13860

ctgctatgtg ccaggtttct gggtgctggg atagaggagt aaacaaggca gagacctggc 13920

ctcagacagc tctcatgggc aacagacaac caaaccaaca aagataccaa gtcatttcat 13980

actgtgatga ggccatagag gaaatgaact ggggatggat agtcaggtgg cttttttttt 14040

tttttttttt tttttttttt ttttgctgtg catgcatcat gtagaaattc ctgggccaga 14100

gatcaaacct gcaccatagc atctacccgg gccactgcag taacaatgct ggatccttaa 14160

cccactgctc cacatgggaa cttaagtggc tcctttagat gagaccacca agaagagccc 14220

tcctgtcatt ctcctgctta aatgtcccca tggcttcctg gcttagcaca gaataagatc 14280

cagaataaga cctgtgcagg accagtcac aatctgcctc tctgacatca cctccttcca 14340

ctctcctcct gaatcacagt gctcctcatg tttgaatggc agcctctttc ttaataacag 14400

gtctgagttc agatagcagc tcccatgaga tgattctgct tactgtccta tttaaagaag 14460

taccgtctac ccacaagtca ctatcccatt gtgttttatt ttcttcatag cacttatcac 14520

tctcttaaat tattttgttt ggttgttttc tgctcacccc tcagcccctg taagctccag 14580

gagagtggag acttccatca tcttgttcac tgctttatgt tcagagcccg agagatgtac 14640

agaagtatct caggcatctg atatcaattg cacagccact tgaactttat tcttgtaaag 14700

tatccagtgg gtgaaaagtt aatttcagca gatgtgaatg ccaggattgg aaatggtttt 14760

gtcaacgtgt caataattag gagtttccat tgtggctcag tagtaacgaa tccaactagt 14820

atccaggagg aggcaggttt gatccctggc tcactcagtg ggttgaggat ccggcattgc 14880

catgagctgt ggtataggtc acagacaggg cttgggtcct gtgttgtggc tgtggtgtag 14940

gctggtatat gccgcagggc ggccctgaaa agaaagcaaa aaagcattac taattaaacg 15000

cttgagttgg aattttgctt tcccataagc catttctctg ggaatggttt gctgaattag 15060

ataggaaaat aactgatcaa ctttatttac tatccaaata agatattgat aattagcttt 15120

cgtgaatctg ggaagacttt agttatcaag ggactagaat tattgagaag cagctaagag 15180

cagcaaaaat ttcttgcaac atttgattct gtagacagaa aaatttctat ggatctagtt 15240

atttaatctt gagaaagatg ttagatttct aatggtgctt cccgagcctt agagccttat 15300

gatggtactg ccatgttgca gaatcaaagt agttaaggag ggcttcttga cagatcaaat 15360

tccactccta aatggctttg acacagctct ctcctgttta ctctacatct gaatagtcgg 15420

atgtaattta gattggtgaa cacatccttt ccctctccat agccaagaga tccttagtaa 15480

ggttcctgct gaagaatagt tctgtggtaa ccaggtaagc aagggttaac tttttagctg 15540

atgctggaca cggtcaccta catgcaatat tgtttctgtg ggaaaatgga gtctgtgatc 15600

tcaacttaca gatacatagc tagaatatag gtggtatggg acttcccatt gtggcacagt 15660

ggaaacgaat ccgactagga gccatgaggt tgcaggttcg atccctggcc tcactcagtg 15720

ggttaaggat ccagtgttgc catgagctgc ggtgtagttc gcagatgcgg cttggatcct 15780

gtgttgctgt gtctgtggtg taggctggca gttgtagctc cgattggacc cctagcctgg 15840

gaacctccat atgccacggg tgtggcccaa aaaataaaaa ataaaaaaaa ttttttttaa 15900

atagaatata ggtagtgtgg agttaataaa tatcacctct tttcctcgcc caggtgttga 15960

agcatgtgag aaatgaaaca aagcatatgg cagatggaga caaaatagca ctttcagttg 16020

aagctagttt tggagactat gttttaaggc ttcctaatac tgaatcctaa ccctccccag 16080

ctacaaccat cctgaatacc caggttctct ccccaaaggg acaaggcaag atcgagaggt 16140

gagtatttcc agtttattcc tctgagagcc ctcagtaccc gtcacttagg gggcagtgaa 16200

taaaggcagg aagggaggcc aggagcctgg cgctcacaga gctgcagcca cgtgaacaca 16260

gcagcaacgc ggagcaagca tctcccctac tgagaggctc tgcgtacatt gtctctacag 16320

ccctggagtg gtgcagtcca gtgtggactg agttaaaagg ggcctaagag aggagggctg 16380

tggattcaca ctttggggac tgcctaacgt cgttgcccat ggttttccag ggaaggccag 16440

ggaagagtga gaaggcaggt gcctagcaga cctgcatccc ttcccggtga ctcttccctt 16500

ccatgtgtcc tcagcagaga agtggagtgg aaagccaggg taggtatcaa caccttcata 16560

aaaaatagca tgagcacaca ctggcaacag ggccctgtgg atgttactct cttgatatct 16620

tgaacccagg aaaagggaga gccgtgtgct cttcctgatt tagaagatag gatttgagac 16680

tctgcagcta attatgtgac cccaggccac tgtcccaatt ttcctaaatt cctgggtagc 16740

cttttataaa atgaatgtaa taggttgaaa cgccatgtaa aaatggcagg aagggagaat 16800

atatttccca tacatatgta aaggacactg gaaactcaaa aataagaaaa taactccact 16860

aagaatggat aatgattgaa acaaatcttt ttttaaaaga agagatatga gtggctaata 16920

ggcacatgaa aagatgctga atgtcattcg tcatcaggag atgcaaatta aaaccctaat 16980

gggagactac taaataccct ttagaatggt taatattaga aagactgatg atgtggagtt 17040

gccatcatgg ctcggcagaa acgaatctga ctagcatcca tgaggatgca ggttcaattc 17100

ccggcctccc tcggtgggtc aaggatctgg tcttgccatg agctgtggtg gagaccacag 17160

acaaggctca gatctgacgt tgctgtagct gtggtgtagg ccagcggcta caactccaat 17220

ctgaccccta acctgggaac ctccatatgc tgcgggtgcg gccctaaaaa gacaaaaatg 17280

aaaagaaaga ctgacaataa caaagtgtta gcacaggtga ggagcagctg gagtgcatat 17340

gttgctgatg ggaatgtaaa acgggacagc tttggaaaac agtctgagag tttatgaaaa 17400

gttaaccatg caccagggtt cccaagtggc ctaccagcta aagatttggc cttgtcacta 17460

ccagagctca ggttctaccc ctggccctag aatttctgca ggccatggat gcagctacca 17520

aaaacagtat taagcatata cctaccatat gatgcagcca ctccatgact aagtatttac 17580

ccaagagaaa tgaaagcata taaccacaca aagacttaca catgaatatt ctcatacctt 17640

tataataatc aaacactaga tcaagccagg ccaaaaaata tcccctccca aatccccata 17700

acattctaca tatataaaat tctagaaaat gcaaactaat ctatagagac ggaaagcaga 17760

tcaggggtta cctcagactg agggtgcgag gaaggatgaa cacaggaaca cttgggaatt 17820

atgagaatgt ttgctatcct gactatggtg atggttccat ggtgtttgcg catgccaaag 17880

ctttaaacag gtgcagtttg catacattac acttcactat aaaagttgta aaagaaagaa 17940

aggacgcaag gagggaagaa gaactgatac ccaggaattt gtatgaaatt tcacaatcat 18000

cctgctgagg gagaaaaacc aggcataagc cagcacctac tgtatgattc cacttctatg 18060

caattctaga aaaagtgaac tgatcaccgg tgacagagag caggtgaggt gttgccccaga 18120

gttgggggtg ggggagggga tggactacag aaaggcaaga ggaaatttgg ggaagtgata 18180

gaagcattgt gtatcttgga gatgggatca aaacggcgga atagtaggat gggagctcaa 18240

cttctctcct caaaacaaca aaatttacaa ctacatgctg agcaatcttc aaccaaattg 18300

actggaaacc ttcaaaaaga tatcctactc caaaggcaaa gaggaggcca catcaagagg 18360

tatgaggggc gattacatga tataaacaac cccatacctc ctgggtggga agccccccac 18420

agagtagaaa ctaactggtt cacagagact cacctacagg agtgagagtg ctgagcccca 18480

catcaaaccc tcaccagcgg ggatctggca ctgggagaag gagcccctgg agcatctggc 18540

attgaagacc agtggggatt gtacgcagga gccccacagg actgggggaa acggagaccc 18600

cattcttaaa aggcgcacat ggattttaac gtgcactggg tcccagggca aagcaaagtc 18660

tccataggaa tctgggtcaa accagactgc agttcttgga ggacctcctg ggaaaatagg 18720

ggtgaatgtg gcttgttgtg ggggaaggac attggaagca aatctctcgg gaatattcag 18780

cagctgcttt tctctggaag tggccatttt gggaaaatct ggcctcaccc atcagtcagt 18840

cagcactgag aagccccagg gcaaacaaca acccaggtgg gatcacagcc ctgcccctca 18900

gtaaacaggc tacctaaaga cccctcaggc actcggctgc ctctaaccccc atccagagac 18960

taagccccac ccaccagagg gattagaagc agctccacct accagtgggc aggcatcagc 19020

ccctcccatc aggaagccta cagcaagccc ccacacagac ttcagccaca aagggggcag 19080

acaccacaag gaagagaggc tacaactcta ttatctggaa aaaggtcacc acaccaaaaa 19140

cctataaaaa tgaaaagaca gagaactata actcagatga gggagaaaga aaaaccccca 19200

gaaaatcagc tcagcgatga gaagattctc agcctccagg aaaaagactt cagactgtcg 19260

atgctgaaga tgatgcaaga cattgaaaat aaactggagg caaagatgga taacttacag 19320

gaaacactga gcaaagagat acaagataca caacatttct taagttttat atcttgaata 19380

taatggtgat ttgacatgac tatacatcta tcaaagctaa ttgaatttta cacttttttt 19440

ttcttcttag ggctgcacct gtggcatatg attacaaagt tgctaaaagt gaccctcaga 19500

gaggcccaga gcggtggtcc acagcgaaag acgacaaagc tcagcagcgc gccttgtggg 19560

gaagcacagt ctcctcagag attctcaaac taccgctgga ggagttccca ttgtggcgca 19620

gtggttaacg aatccgacta ggaaccacga ggttgcaggt tgatctttgg ccttgctcag 19680

tgggttaagg atccggcgtg gctgtgagct gtggtgtagg tggcagacgc ggctcggatc 19740

ccgcattgct gtggctgtgg tgtaggctgg cggctacagc tccgatgaga cccctagcct 19800

gggaacctcc acatgccaca ggagccgccc tagaaaaggc aaaaaactac cgctggagga 19860

aacaggatgt cctgcatccc agtggagtga ctggagcaca aactctgtga ttgggtattt 19920

gaaggaattg tgacttccaa gaagatggta gcttggggtg tttggaatgt gaagagatta 19980

gctgacaccc ccgccccctt ccgctgtccc ccaaccccca tcctcccaag gacgtttggg 20040

ctgtgttgag ttattttata ctatgagaca gaaaaataat ctataagact agttttgcct 20100

cttccatcac aaaagctcag aagagaccta cgctctatga cccaacccta actagcagtt 20160

tcctttgtct ggtttgacgt cagttaatga ctggactttg aacctgaaag aacattaaaa 20220

ggaaagatag ctgaagggaa aattttctct ctggattacc tggcaccaac atccaaaatg 20280

aaagtcttca ttcacttcac tctgcttgcc ctccttttcc taaaatcagg taaggaacat 20340

ttacacattt ttaattcttt ccgtcatatt tgtctttgag atagaattag caggcttaag 20400

ctaccaatcc cctctgcccc aactagtggg ctattttgta gatttcctcg ccaaccaggc 20460

tcaaagatcc aagccaggga gttcctgttg tggctaagtg ggttacaaac ctgactagta 20520

gtctatgagg acatggattc aatccatggc ctagctcagt gggttaaaga tctggcattg 20580

ctgtgagtgg cagtgtaggt tgcagacatg gctcggatgc tgcattgctg tggctgtggc 20640

gtgggccagt agctgtagct ctgattcaac ctggagcctg ggaacctccg tatgtgaccc 20700

taaaaagaaa aataatgaag aagaagaaga cacaaggcat ttcaacctgt gatgaaagca 20760

ggtagaacat tagtaagagg ggcttctgga ctaagatttg gtgttcagca aggcagacat 20820

tttcttgagt gtcctgaggc cagacaggaa cagcttagta attccagggc ccaatgcctt 20880

agagtctcac taaccactag atatcacttg ggttctaatg cagagtaggg atctggtttg 20940

atgcctgaat cttgcatcaa gagttttctg cccttgctgc cctgttgtgg cgcagaagaa 21000

actcatccga gtttgatgcc tgaatcttgc atcaagagtt ttctgccctt gctgccctgt 21060

tgtggcacag cagaaactca tccgacttgc aaccatgagg ttgagggttc aatccctggc 21120

ttcgctcagt gggttaagga tctggcattg ctatgagctg tggtgtaggt cagagatgca 21180

gctcggatct ggcgttgctg tggctgtggc gtaggctggc agctgtagct ctgattatagac 21240

ccctagcctg ggaacctcca tatgctgcag gcatggtcct aaaaagacaa aaaacaaaac 21300

aacaaaaaaa gtgttttctg ccctttacac gtgggaattg aggcttatct ctggacaaat 21360

cctttagttc ctctgagtat tggtgggcat ttctgtcaaa tgtggatgct cacatccatt 21420

ccataggcac tgttgggcac agggtaagtc cttgataaat gataggagga gtgggaacag 21480

gggcagcagc agtcaaggtg ctcagctggt gaactatttt ctagccccta gaagaaaact 21540

ggcttatgaa tgatggttta agccagaccc tgttcaaggt gaagagccaa attctgctac 21600

ccagaggctt ccacacccaa gggggaatca ggtagcttca gcattttttc tatgtctgaa 21660

agtttaggga tagcttcctc attaaaactt atcttttaat attttaaacc caccttcatt 21720

gtcagaattc ttagagacaa agtttctaaa aaagtgactg gatcacaaaa agcttattac 21780

cccatatgtc caggggttcc taagagttga aaaaaggcaa ctttggaagt tcccactgtg 21840

gcgcaatggg ataggtggta tctctgcagc tctgggacac aggttcgatc ccccccagcc 21900

accgtgggct aaggatctgg cataggttgc aactgcagct tggatcttat ccctggcctg 21960

ggaactccat ttgccatggg ggaccctcaa aaaaggcaag tttgaggatc accatagccc 22020

ttagagtaaa gatcatctct atcattttac tctttaaaaa cttcatgtgt ctagaattat 22080

aaatgggtaa ataacactag aaaaaaattt gaaagaagtt taatacacac ttaatgtatt 22140

aacgattttt atttaatcaa tctttaaaca catccccttc cctaccatcc ctcagctatt 22200

atattgtgta atggaaaatt ggggcaaatg ccttaaagtg taaaaggaca aaaggaggcc 22260

atttccaggt agtaacagtg ccaccttgtg gcattttcaa ttttcctcac tgagaatcat 22320

ggaggaatat aaactcagat tgacagatct gcaactatag gcataaatga tactacatct 22380

tgtgtcaaca tctgatatgt tgatattaat atgtatatat aattatactt tatatatcat 22440

aatatcatat atcatattat aaccaataat taatacaatt aataataaga taaatgatgt 22500

taatacttct ttatccagtt gagttaacag accaggtacc atcatatttt ccagtaaccc 22560

ttttccagtt ttgtctgttt aattttttct ttgtctttgc cttttctagg gccgctctgg 22620

cgacatatgg aggttcccag ggtaggggtc gaataggagc tgtagccacc ggccttcgac 22680

agagccacag caacacggga tccaagccat gtctgcaacc tacaccacag ctcacggcaa 22740

tgctggatcc ttaacccact gaacaaggcc agggatggaa ccctcaacct catggttcct 22800

agtcggattc gttaaccact gccccacgac gggaactcca attttttatt tttctttatg 22860

tgtttttttt tttagatgta gtcatgtatt ttcattttta atgtactcca ctacatctac 22920

ttttcagaaa aaaaaaggat tgggtaccaa ttagtattac atagtagcct ctgccattat 22980

atatacagtg ttttgtcttc tgaaactttg cctttcagca agtatggctc agaacaagac 23040

aggaggcacc acaaagctga catccacaga aaacaccagc tccccgaacg cggcggaaag 23100

caccagcgct ccttccgctg caagaaacag tgctccgggc acggcagcaa gcaccagcgc 23160

tccgtctgcg gcagcaagca ccagcgctcc ggccgcaggg gcaagcacca gcgctccggc 23220

cgcaggggca agcaccagcg ctccgtctgt ggcggcaagc accagcgctc cggccgcagg 23280

ggcaagcacc agcgctcctt ccgctgcaag cacgagcgca ccggccgcag gggcaagcac 23340

cagcgctcca gccgctgcgg caagcaccag cgctccagcc cccgcggcaa gcaccagcgc 23400

tgccgcggca tcaagcacca gcgctccggc cgtggcggca agcaccagcg ctccggcccc 23460

cgcggcaagc accagcgctc cctccgctgc ggcaagcacc agtgctccgg ccgcaggggc 23520

aagcaccagc gctccggacc ccacggcaag caccagcgct caggccgcag aggcaagcac 23580

aagcgctccg tcggcggcgg caagcaccag cgctccggcc gcaggggcaa gcaccagcgc 23640

tccgtctgtg gcggcaagca ccagcactcc ggccgcaggg gcaagcacca gcggtccggc 23700

cgccacggca agcaccagcg ctccggccac aggggcaagc accagcgctc cgtctgtggc 23760

ggcaagcacc agcactccgg ccgcaggggc aagcaccagc ggtccggccg ccgcggcaag 23820

caccagcgct ccggccgcag gggcaagcac cagcgctccg tctgtggcgg caagcaccag 23880

cactccggcc gcaggggcaa gcaccagcgg tccggccgcc gcggcaagca ccagcgctcc 23940

ggccacaggg gcaagcacca gcggtccggc cgcggcggca agcaccagcg ctccggctgt 24000

aggggcaagc accagcggtc cagccgcgtc agcaagcacc agcgctctgt ctgaggcgga 24060

aagcaccagc tctccagccg caggggcaag caccagcgct ccgtctgtgg cggcaagcac 24120

cagcgctcca gccgcggcgg caagcaccag cgctccttcc gctgcaagca ccagcagtcc 24180

ggaccccacg gcaagcacca gcgctccggc cgcaggggca agcaccagcg ctccctccgc 24240

tgcggcaagc accagcgctc cggccgcagg ggcaagcacc agcgctccct ccgctgcggc 24300

aagcaccagc tctccggccg caggggcaag caccagcgct ccttctgctg caagcaccag 24360

cgctccgtct gtggcggcaa gcaccagcgc tccggcccct gtggcaagca ccagcgctcc 24420

ggccccaggg gcaagcacca gcgctccttc tgctgcaagc accagcgctc cggccacagg 24480

ggcaagcacc agtgctccct ccgctgcggc aagcaccagc gctccgtcca cggcggcaag 24540

caccagcgct ccggccgcag gggcaagcac cagcgctccg gcccccgcgg caagcaccag 24600

cgctccggcc cccgcggcaa gcaccagcgc tccagccccc gcggcaagca ccagcgctcc 24660

ggctataggg gcaagcacca gcgctccgtc cgctgcaagc accagcgcac cgtctgctgc 24720

aagcaccagc gctccggccg caggggcaag caccagcgct ccgtctgtgg cggcaagcac 24780

cagcgctccg gcccctgcgg caagcaccag cgctccggcc gcaggggcaa gcaccagcgc 24840

tccttccgct gcaagcacca gcgctccagc cgcaggggca agcaccagcg ctccggccgc 24900

ggcggcaagc accagcactc cagaccccac ggcaagcacc agcgctccgg ccgcaggggc 24960

aagcaccagc gctccggccc ctgcggcaag ccccagcgct ccggccgcag ggacaagcac 25020

cagcggtcca gcccccgcgg caagcaccag cgctccggcc gccgcggcaa gcaccagccc 25080

tctggaccccc acggcaagca ccagcactcc ggccgcaggg gcaagcacca gcgctccatc 25140

ggcagcggca agcaccagcg ctccttctgc tgcaagcacc agcgctcctt ctgctgcaag 25200

caccagcgct ccggccgcag gggcaagcac cagcgctccg gccgtggagg caagcaccag 25260

cgctccggcc cccgcggcaa gcaccagcgc tccggctgca ggggcaagca ccagcgctcc 25320

gtccgctgca agcaccagcg ctccggccgc aggggcaagc accagcgctc cggccgcggc 25380

agcaagcacc agcgctccgg cccccacggc aagcaccagc gctcccgccg cggtggcaag 25440

caccagcgct gcggcccccg cggcaaccac cagcgctctg gccgcggtgg caagcaccaa 25500

ccctccggcc gccggggcaa gcaccagctc tccggccgca ggggcaagca ccagcgctcc 25560

gtctgtggcg gcaagcacca gcgctccggc ccctgtggca agcaccagcg ctccggccgc 25620

aggggcaagc accagcgctc cttccgctgc aagcaccagc gctccggccg caggggcaag 25680

caccagcgct ccggcccccg gggcaagcac cagcgctccg gcccccgcgg caagcaccag 25740

cgctccttcc gctgcaagca ccagcgctcc ggccataggg gcaagcacca gcggtccagc 25800

ccccgcggca agcaccagcg ctccggccgc cgcggcaagc accagcgctc cggacccccac 25860

ggcaagcacc agcactccgg ccgcaggggc aagcaccagc gctccgtcgg cagcggcaag 25920

caccagcgct ccggcccccg cggcaagcac cagcgctcct tctgctgcaa gcaccagcgc 25980

tctggaccccc acggcaagca ccagcgctcc gtccgctgca agcaccagcg ctccggccgc 26040

aggggcaagc accagcgctc cggcccccgc ggcaagcacc agcactccgg ctgcaggggc 26100

aagcaccagc gctccttctg ctgcaagcac cagcgctccg gccgcagggg caagcaccag 26160

cgctccggcc cccacggcaa gcaccagcgc tcccgccgcg gtggcaagca ccagcgctgc 26220

ggcccccacg gcaagcacca gcgctccggc cgcggcagca agcaccagca ctctggcccc 26280

cacggcaagc accagcgctc cggctcccgc ggcaagcacc agtgctccgt ccgcggcaag 26340

cacaagcgct ccggccgcgg cggcaagcac cagcgctccg gcccccgcgg caagcaccag 26400

cgctccggct ggggcggcaa gcaccagcgc tccggctgca ggggcaagca ccagcgctcc 26460

atccgctgca agcaccagcg ctccggccgc ggcggcaagc accagcgctc cggccgcagg 26520

ggcaagcacc agcgctccgg ccgccgcggc aagcaccagc gctccggccc ccgctgcaag 26580

caccagcgct ccggcggcgg tggcaagcac cagcgctccg gccccagggg caagcaccag 26640

cgctccatcc gtggcaagca ccagcgctcc ggcccccgct gcaagcacca gctctccggc 26700

cgcaggggca agcaccagcg ctccatccgc ggcaagcacc agcgctccgg tccccgctgc 26760

aagcaccagc tctccggccg caggggcaag caccagcgct ccggcccccg ctgcaagcac 26820

cagctctccg gctgcagggg caagcaccag cgctccggcc gcgggggcaa gcaccagcgc 26880

tccggccgca ggggcaagca ccagcgctcc agcccccgct gcaagcacca gctctccggc 26940

cgcaggggca agcaccagcg ctccggcccc cgctgcaagc accagcgctg cggccgcggg 27000

ggcaagcacc agcgctccat ccgcggaaag caccagcgct ccggctgcag gggcaagcac 27060

cagcgctccg gctgcagggg caagcaccag tgctccggcc gcgggggcaa gcaccagcgc 27120

tccatccgcg gcaagcacca gcgctccggc cgccgcagca agcaccactc ctgcctctgc 27180

agcaaatgct tcttctgcta tttcgagtac acctattgcc tccacaccca cctcaggtaa 27240

gagagattcc ctcacatctc ctcactgagg catcagggcc ttttggcagt tgacaggtac 27300

gtgtgttgtt cgggaatggg gaagtgcata taaagggacc caagaggacc ttggttacga 27360

aaccctggag tcacaaatgc taatagtgat ggcaaagatg ctaaaagatt acctgcagtt 27420

ccatgctatt cccctttcct caagcatttg ccgtatgtgg aatattccta tgagttggac 27480

tttatgaaag ctcctgttcc agcttccttc tccctaggag atttttggaa ataatgtcag 27540

cttctttgag aatatcaggt ccaaataaac actgactctt cagccccact gatctagtga 27600

ctgaaggaga gagtcacttc tactctgatt ccacatcacg tgttattgaa aaatatgaca 27660

ttaaatggta ggtgatcaca atttggtatt tgagctaatt catggaattt gaggctaagg 27720

aataaaagcc aaacaagagc attgctcaaa gagtcctaga catgaccaag gccatggtcc 27780

aaaagacgga tgtttgggga tacttatcta agacaagatg attgatctca agccaaagtt 27840

gaaaccaaaa ccacttgcgt catttcccaa agtttggtgt agagcaagtg tcaataaagt 27900

ttatcagtga aaggaaacta ataagtagtt taggttttat gggccgtagt aaggtctttg 27960

ttacaactac tccactctgt cctgttaaca tcaaaacagc cataggcaat atacaaatga 28020

atgtgcctga ctgtgtgcca ataaaacttt atttgcaaaa tcagaaaact ggtcagaatt 28080

ggcactcaag tcatcatttg ccaaaccctt gtctaaagga atgctgaaac ctaggcttca 28140

agggtctata taaaccctgt tattccaatt taggggaact gaagtccagt aagggcagca 28200

acttgcccaa ggccaaaagt caagtcgatg tctccctgag cttatttctt gtggcttccc 28260

caatgttctt tgatagacct gtgaaatgaa tgattaagca aatggagcca gagcctgtat 28320

caaaacttgg ttcccaggct tcttttgtgt ttgaagggtc cactctgaag gccaggtcaa 28380

gagaaccaca gctgcacaat cagttctcca gtagggagct tatctaagct gttttgcttc 28440

cctagacact cttgcagagt aaggcagtct tcttgtgttc tggccctaca ctaactaact 28500

ctggggcaca aggcaaatca cttctgtctt ggcttcaatg gactcatctg taacaaggtg 28560

tgactgaact gaatagccat cccggaattc ctgttgtggc tcaacaggtt aagaacccaa 28620

ctagtatcca cgaggatgtg ggctctatca gtgggttaag gatctggcat tgctgcaagc 28680

tgtggcatag gatgtggcta ggatcctgtg ttgctgtggc tatactttaa gccagcagct 28740

acagctctga ttcaaccccc atggcttggg aacctccata tgccatgagt gcagccctaa 28800

aaagacaaaa aaaaatagcc acccaaatcc ttttcccact gtgacatcct acgactccat 28860

tcctggcaca cagaaatgtt aacaagtctg ttctttatg ccagggagaa agcacaccgg 28920

gagttatctg tctggtatat tcagaatgga gctgtactaa atttcttccc tagaattttc 28980

caatgtgact gaggcttttt taaaaaagac atttacttta cttagaatgt tggtttacat 29040

cttctcaaag gggtgaccct cacatggttt atgagatctt gtgccatgcc tcttatgtcc 29100

catagcatgg ggtttgggat ttgggggttg attttgtgtg ggtttttttt cccccttaaa 29160

atagagcaat tttgggttca cagcaaagtt gagaggaagg tacatagatt tcccatgttt 29220

cccggtcctt gtgcctgtac agcctccctt atcatcaaca tccctcgcca gatggcacat 29280

ttgtgacaac tgatgactct acattgacat attataatcg cccaagtcca gtttacgtta 29340

gggtttacag ttgatgttgt atgttctata ggtttcaaca agcatatggt gacatgcatc 29400

catcattacg ctatcgtaaa gactgttttc acctgcttat tcatcctccc tgctaaccct 29460

tggcaaccac taatcttttt tactattgcc atagttttgc ctttttcaga atgtcatgta 29520

gttggaatcc cacagtatgc agccttttca gattggcatc tttcatttag caatggcatt 29580

tgagtcctcc atgtcagttt caaggcttga gcactgagta atattccatt gtctgattat 29640

accagtttgt ttatccactc acctactgaa ggacatcttg gttgcttcca aattttggca 29700

gttatgaata caatgctcta agtgtttgta tacagggttt tgagtggata taagttggta 29760

aatacttagt tacatcaggt aaattacaag acaggcactt gctaggtcat gtggttaaga 29820

gtgtgtttag ttttgtaaga aactgccaac atgtcttcta aagtggctga accatttcac 29880

atttgagagt tcctcaccag cagtatttta gtccataggt tataagaaaa gaagcttaaa 29940

gagggcaaaa gtatatgaaa taaaaatgaa atcttcctcc tatcctaaac tcctaggacc 30000

cagacttcag tttcaacttc tgcagccaat tttttgtgtt tgtgcaaata caaacgaata 30060

tacttgtatg tttctgtgta ttcctctcct tttttttaat catgaaaaag tgcagggtat 30120

ttctggccat atacccaaga gtggaattgc tggatcatag ggtaaaacta aaaataaact 30180

accctgtgat ccagcaattc cattccttgg tatatagcca ggaaaaaaaa atgaaaacac 30240

taacttgaaa agatccacgc acccccatgt tcataagagc actacttaca atagccaaga 30300

catgggagca acccaagtgc ccaccaatag actggataaa gaaaatgtga tatgtgtgtg 30360

tgtgtgtggc tgtaatgata gtacttatat gtagaatcta aaaaaataat actggagttc 30420

ctgttgtgac tcagtggtta acgaatccga ctaggaacca tgaggttgca ggtttggtcc 30480

ctggccttgc tcagtggttt aaggatccgg tgttgccgtg aactgtagtg taggtcgcag 30540

atgcggctcg gatcccacat tgctgtggct ctggcgtagg ccggcagcta cagctcagat 30600

tagacccccta gcctgggatc ctccatatgt ggtggggaaa ggccctagaa aagacaataa 30660

ataaataaat taataataca aatgagtgta tatacaaaac aaaaacagac tcacagatag 30720

agaaagtaaa cttgtagtta ccaaagggaa gacagacaaa ttaggggtat gggattaaca 30780

gaaacaaact actatgcata aaatagagaa gcaacaaagg taaattgtat agcacaggaa 30840

attttagrtg ttatcttata ataacttgta rtatartctg caaaaatact taatcctatg 30900

ctgtatgctt gamactrata magtactgta ratcaatgat acttcaataa aaagagacca 30960

tgttggagtt cccatcgtgg ctcagtggaa acaaatctga ctagcatcca tgaggatgca 31020

ggttaaatcc ctggccttgc tcaatgggtt aaggatccgg cattgccgtg agctgtggtg 31080

taggtcacag acacggcttg gatttggcgt tgtcgtggct gtgacataga ccagtgttta 31140

tagttccgat ttgaccccta gcccaggaac ctctgtatgc cactggtgag gacctaaaaa 31200

aaaaaaaaaa aagagagaga gggagaccat gttataccca ttttagttta tgtatagctt 31260

tttttttttt aagcaatgat ttccaccacat aggatcctaa gaatcttaag acmgcactga 31320

gggttgtcag tcacttgatt ctatcctgaa accaaagttc agaatcattt cacttaacaa 31380

agataaagaa aatggctaac caagctcatc ttttggccac tgccctaact tttccttctc 31440

agatcccatc aattctccag agacaggcaa gatgctgcat cctgcctggt ctaggagtcc 31500

cagcccatgg tgtagtaacc attgtaatgt tcttctcacc atttattcag gtcatccacc 31560

caaaaaacca gcgatcccct catcacttcc tcttagaatc ttcactggat taatccactt 31620

tgttccctta ctatcccttt cttttcttcg taggtctgta taccatgcat actaaccaaa 31680

atataacacc taatytattt ttggcagctt cttgcctagt cctctctgta agtcagttgt 31740

tcacaaaata tgatgctttc tctgaaatgg ttttagggaa cactacctag tgttagaaaa 31800

ccagatgtca aggttctgga ggatccttga ggtccaaatt atttccttat tcctcagaat 31860

tagaagatgc cctctctctc tctgcacaga ctcatccacc tctgatcttt cctgtttgac 31920

aagtgcttgg ctccaaaatt ttagcaaaaa actttccaga gacataagaa aatatgtact 31980

aacagaaatt gacagaacat tgtaaatcaa ctgtaacaaa aaaagaaaaa gaaaaagaaa 32040

gaaaatatgt atttgtttct agctgatta agggtgtgcc atggagtaag acagctgaag 32100

aatagaaagt gttcaggaga ggaggcatga atatttaaaa gcatgtttct gggtgctgtt 32160

tgtttgttgt cattctgtgt atttttaacg tttttctact ccattgcagg ttccaggaac 32220

ccttgccagc aagattcctg taagggtggt tcttcatgtg ttagtctgaa taatacgttt 32280

ttttgcctgt gttcagaagg gtattactat aactcttcca cctgcaataa aggtgagtta 32340

gaagggatga atcaggtaga gctgcatctc agacctaaaa tagaagagaa aacttaaact 32400

aatagcaaga agaacttcta tagtattaga ggcagatttg gaggcaacaa agcccaggag 32460

agtacagaaa agagtcccgt cttatttcag tgcaaactga tgggagagga aatggactaa 32520

gtcagatttg tgagctagtc ctaaaggtgt gcgtgtggat gggtcccttc acctgacctg 32580

ttcaagggaa tggtgccatg ggatcaaagt cagaatgata ggtgttgacg tccctggaca 32640

ggtacaaagg tgtataaata ttaagtgatg ctgttacatg gggacactga tgcagaatta 32700

aggtgatttc ctaaaaccac acagaccagc cggaaacctg gtctctgacc ttggttcagt 32760

tctctgtcct aagatttgca taggaaatga aagcattgta ttggccacaa ggattagata 32820

tcccaaaagc caaaatagat gttcatrttt ttagcactaa taatctcatt ttggggatcc 32880

aacactaagtc aatgacccta aatacgggaa gttttatgca cgaagaggtt cattgcagtt 32940

ttttctcttt tragtagtta aatgcactga aaacaagaaa tgactgcatt ggagytcctt 33000

ggtggctcag caggttaaag atctgttgtc actgctgtgg ctcaggtcra tgcagtggtg 33060

crggttcagt ccctggtctg ggaatttcct cttrttatgg gtgtagccaa aagagaaaaa 33120

atgaaacaaa acaaaayaaa aatgacttca tcgataaagg aaacatgtac ttcctagaat 33180

gttttgtagc atttaaaatg ataattatga agtctaagta atgatttggg aaatgcttat 33240

gtaatatacr aagacatgat ataaacttgg arttataatg atgtaaacta gggacacaaa 33300

aaagctggga gaaatatgcc aaaatgagga tagttgtggt aggttaggtg rgggttgggg 33360

tcatgttttt cmtattcmca gacttcctgt agtatcatca gacatttaga atttttaaaa 33420

aattatctgt tttaaagaaa agagaaatgc ctctcttcac attaaatatc acagaaagat 33480

tgtgtttgaa tttcaggaaa gattttccct ggtaaaatta cggtcaaaat atcagaaaca 33540

actgatctag aacatgaaaa ttctgtgacc tatcaaaatt tataccttca agttacaaat 33600

tttgtaagta ttttccaacc atccttccta tacaaatatt ctttatatat atatatataa 33660

tgatttctak ttttttccca ttatagctgg tttacagtgt tctgtcaatt ttctgctgta 33720

cagcatggtg accccagttac acatacacgc atacgttctt ttttctcaca ttatcatgct 33780

ccatcataag tggccagaca tagttcccag tgttacacag aagtagctta ttgcttatcc 33840

attccaaagg caatagtctg tgtctgttaa ccccaagcac ccatccatcc tactccctcc 33900

cctgatacta tatatagaaa accctaagga ctcaacccag aaactacctg aactgatcaa 33960

caaattcaga aaagtagtag ggtataagat taatattcat aaatcagtca catttctgta 34020

tactaacaat gaaatattag aaaaggaata taaaaatata atacctttta aaattgcacc 34080

ccaaaaaatc aaatacctgg ccaagaacrt aaaagactta tatgctgaga gctataaaac 34140

atgaatcaag gaaatcaaat aagatgtgaa gaaatggaaa gctattccrt gctcctgggt 34200

tggaaaaatt aatatygtaa aaatggccat actacccaaa gcaatctaca gatccaatgc 34260

atccctatca aattacccat gayatttttc mcagaactag rrsraacaaa caatccaaaa 34320

acttatgtgg aaccacaaaa gaccyagaat taccaaggca attccgarga acaaaaacca 34380

agkaggaggc ataactctcc cagacttcag gcaataaata ttctttaact gacttgttag 34440

actgatattc ggrtggtaca ggctggtacc agtgattcam ataggtgaaa tgcatcctta 34500

ttggttggtg aagagcagct gttccaaact cttgggtgcy gtggcttcct ggttgcctag 34560

actccacctc tgggaccctg ggacctcccc rtgatcctac acctagagtg taacacccag 34620

cactgcagag agcctttaga agcctgctat agccatgcta tacttgtttt tatggaggct 34680

attaaaatat ttttkaatgg acagataaat taattatact trrraataag ttatttccat 34740

agatttgaag agaaaaaaat tatttgatta ctcaatgaat ttatrcatt tatagttgat 34800

catcacaatc cagttttata gtgtttccat cccayacccc cagcgcatcc ccccaccctc 34860

caacctctct cctttggaaa ccataagttt ttcaaagtct gtgagtcagt acctattctg 34920

caaagaagtt ccttgygtcc ttttttcaga ttcccgamct gtcagtgaaa gcattcaatg 34980

ttggtgtctc attgtctgac tgacgtcact yagcatgatc atttctaggt ccgtccatgt 35040

tgstwaaaat gccggtattt cattcctttt aatggctgag taatattcca ttgtgtatat 35100

gtaccacmtc ttctkgatcc actcctctgt ygatggacat ttaggttgtt yccatgtctt 35160

ggctattgca aacagtgctg caatgaacat cggagtacat gtgtcttttt gagtcatggt 35220

tttctctgga tagatgccca ggagtgggat tgctggatca aatggtagtt ctatgtttag 35280

ttttctgagg aatctcccca ctgttttcca cagtggttgc accaatttac aatccccacca 35340

agagtgtact agggttcctt tttctccaca ccctctccag cacttattgt ttgtagactt 35400

tttgatgatg gccattctgr cyrgtgtaag gtggtacctc atagtggttt tgatttgcat 35460

tttgaagaga aaaaaattta aaggtgttgt tttttccatt ttaccatcca atttgcatac 35520

atttctagac tagtccagct tttttaatgt cygaaggaac tccttctaaa ttaacatccg 35580

ttacttcttt ttcccattgt caaaaatgtc caaatgagca acagcttcat ttctttgtga 35640

ttattgtgag tgaatrtgtc gatgtgagag aggtggagat ctctattcta aaaccacagc 35700

tgtcagacag gaaggcctta gagaaggtgt cycatcrgtg tttctcaaar tttagttttg 35760

aaggaaaaaa tgttagcttg gaaatatgtc taagcacata attctaggtg aagaaatgct 35820

ttttaaaack rrsatcttac tctatgatac tgcctttccg tttgacatta tgtcattgta 35880

aagagtttta ttgmtagaat tggctaaaag tgaaaartta ttatccaaat ggtcttagag 35940

attgtacata taaagtgagt gctcacaacc aaacagccag tgcctgacac tgctctgata 36000

aaacccatgt ccactgcatc ccggacttgc cctggcccct ttgctgttyc acctgcacat 36060

gaggggcrca tggccatgac ttagcytctt accctgggtc tgcattccag ttccccactc 36120

agaagatatt cagtacattg aagatagcaa cacratgttc ctgtctccag ggaacccctc 36180

aattgtctta tgtracttcc ccaakccctg gtccctggag ccactgcatt tcccaacttg 36240

ctttggatct gaatgtccaa agcccygtct ggcttcaggc caacgccctg gtcttcggag 36300

ataagtgtag caacytgatc tcagatagga cttgatggtc ccaggggtac cagggctggg 36360

atggaacact ttcataattt ctgagagtct ttgaatctga aagggatctg ccagaaygcc 36420

tttgcagctc ctagagagag gaaaagagag acaattgact ttcttcaagg ycccctttgc 36480

agactctgta gctgtggcag tgccagctca ggccctgagg ggtctcccag ggcccatttt 36540

ctccccawgc agagccaaat ggacagtcca catagggakt ttttttttaa tgagtcatat 36600

tggagtatag ttgacttaca aagagttaat ttcaggtata cagcaaagtg aatcagccat 36660

acatatacat atacatatat atccattcct tttcagattc tttccccaaa taggtcacca 36720

cagggtactg agtaaattcc ctgtgctgta cagtagatcc tggttactta ctgattttgt 36780

acatagtagc gtgtctgggc caatcgcaac ctccccagttt atccctcccc tcaatgtttc 36840

cccataatca gagggatttg aagtttccct gttgcatgcc atacctggca ttgcatgtgt 36900

catatttcta attctcaaaa taggcacctg ggggagacgg tactttgata caggtttgca 36960

ggtgaggtag gtagtcaagg atcaggaagg ctatgtaact gaataactta cggaaggtca 37020

cccagctgac gaatgaggat ggggagatgg gagcccagga cctgcctgac cacagagccg 37080

cttctycttc tctcctgctg gctctcttgt gccatggact ccagacaggc ctgtttggat 37140

agttcctgcc ctcctgtcca accctgcagc ccttcacatt gaaaggtgta tagccttaaa 37200

tcctcagagg gtkacctgga agagggggaa aaaaaggaaa tcttttcttc cctgtgcccg 37260

atgctaattt trcaatccct tctttggaat taagaaaact aaatgggcaa tggtcagtag 37320

gsgagctgta aggaaaactt ggggtggaac ttgttccagt taatttgttc ctaagactcc 37380

cagggccctg agggtttata ggaataccag ctttccctga tattkaaaag aaktaaaatg 37440

aatatggaca tattctggat acaaggctkc ccagactaac taaatttatt tgaatttgac 37500

taaaataaaa aagaaatgat ttgatatctt tttaaatgac attggctgat cttctaaaat 37560

rtggggtcct tgtggggagaa aaataataaa attcttgggt tataaaaaat ttgcgccaac 37620

catgttcaac attgacataa taaatatggr gaccakggcc caaagagaag attttccaa 37680

agtcatgtct ttcatagtta attcattcag tcatcagtca actggtacct gttgagtgtt 37740

tcatggggat tgccctgtgc tcagtgctat gggtgggaat gggtgacaga caggtgtatg 37800

catgtggggg gtgtgtcatg agaaagagga aattaactcc agcctgacat caagaaactt 37860

atgttccaac tgggacccct gccctacctc cacctcaccc ctactagaag tcccagctcc 37920

gtctctggcc atgtctttat aagtatctag agttgtctct gccttttttt ttagggccac 37980

acttgcagca tacggaagat cccttgctag gggtcgaatc ggagctgtca ctgccagcct 38040

acaccacaga cacagcaaca ctagatccaa gccacatctg caacctacac tgcagctcac 38100

agcatygtca gatccttaac ccactgatca aggccaggga tcgaactctt atcttcatgg 38160

ayactagtca ggtccwctac caccaagcca caatgggaac tcccaagttg tctctgctct 38220

tgaatacaac tcaatcccaa taagatcaac ttgtgctgga gcccttccca gttcccattt 38280

cacaaagcca tgtatcatat ataccatctc atttcctaca tcgtatagcc ctatgtggtg 38340

ggaattactg atctctcaga ttttccaaga ggaagtgaag cccaaagaag ttaaatttwg 38400

carcaaagat gacaagttgt gtaatgtgac actccagcct gctaccttcy gagtggcact 38460

gtttcccagt gttggtgtgg gtggctcctt ctwctgcttc ctggccccac atgaytctcg 38520

aaatgcttta agtcccatct cctttatgaa gccctctcyg gtttkctaca actggaagtg 38580

agtttctttg aacttccaca ttagattgta ctctctgtgg cacttacctc tctcagctcc 38640

atggtgtaac ctcctatttg atcacacgct gggtctgccc atcttggtta tctctatgtt 38700

tctggcatga tgcctcagca aggtaggcaa ctgagtcttc ctttggtaaa ctgatgagcg 38760

tatgaagtac gcrtaaagat aaattaacaa atgtgcagaa cttcacagtg gctttgaaaa 38820

gacaaaagtt agtcacatta tttttaagac tgtgatcatt gatttggctg agtgctttga 38880

gtttccccct tcttcctggg gagctctgca ttgtatcagg atcktactac tttgaagcaa 38940

aaaaatactg aacgttktta gcgtgtgtgt gaagagtaat ttgctgtwaw awaaawgtta 39000

attrwawaw waatgwwaaw awrwawawrw awawwrwwaw awaaatgttt agaaatarat 39060

ttatgttaaa attaaaatat agcatcacgt ttccccctct ttgtagttac taaatctgtt 39120

cttcttctcc acagtttaat gacacattta gcaattctga gtacggacag accgtaattc 39180

ttaaagtaag gtaagtttac tgtggaccga tgrgktactt tggaagcagg tgtgagtttg 39240

cctaacttcc tgtcttctct tgcattcccc atgcagcacc aaggccctag ggtggctgct 39300

cacatgttaa gaagaagtag tcagtcagag gactgcaggg gsccctgcct ctgatgtcac 39360

attgacccca gcctgggtgg gtggggtgaa gcctgggaca gcaagttgct gtctgtaatt 39420

ggctgctcag cgacttacac acatggtcta gggcatcacc tggatcggtt tggttggttt 39480

gatcccttgg ctacaaacca cacctagtcc acccctggaa tgtgcattcc aggagtcag 39540

gaggcgaatg aatggctcac ggccatctct tccctcatag cggggagtat ggcagacgac 39600

aggaaggtat cagtcatgca cctggctact gaaggccctg tcgtgagctc tttctgccat 39660

gagagtaagc atgcaatgag cccagcaaca ttcagtgacc aggactgcat gctatttggg 39720

gacccaaaaa ggcagaggac tctcaagaag tcagtgattc catctcagcc trccctgaag 39780

casattgata cacaagggct gcagtctgag gaaagaatgt cctaggatrc atggagtcat 39840

ctagtactgc agccctaaca tttctgtttt acggttttac tactgcatct ctctttgtaa 39900

actgtgtcga atcctattgt ggaaaaagat aatctgtata tacttagata tagtatatat 39960

ctatatasag aaggcctcct aaatatggtg agytttgatc ttgrtgrtga asgaggggca 40020

cttgtgattg ggagccagga ggccagcakg gagcgtartg ggatgcacag akcaaacatc 40080

aggcagggag gctgggctct gactgcagag gctggaagcg ggagggatag cagaccactg 40140

tcatgtagtg tttaaggacg ttagtttggt tgaggggacg acagggtgtt catacatcgg 40200

gttgaccagg taaaaacagg aatggttagg tgtgggtgtg tccgctaaat ctgtacttga 40260

atcctggcct cagcattttc ttgctgtggg accgggcagg cattttcacc tctctgaact 40320

ttcgatacct cttctgcatg actgggatag tctgatcaac tttcccggaa ccctgggaga 40380

agcgaatgag ataccattga aaagatgccc tgtgtaaagt ttctggcggg tagtcagttt 40440

cagtatgtgt tcatttccat cagtttccct tccagaatac cagatcctac aagtgtgatg 40500

gagagctggt cccacatggt cacttgggga gccggggggg gggggtcct taaaatagtg 40560

tccctacttt cattcttaag gagttactac tgaaaggacg cccatggagt ttggaagcaa 40620

ccccttgcct gtgagctcag ggtcctggag aacacccacct tcccccagct tgggggctga 40680

ggagggcaac gagacctcag tctcacctca gaagacattt ctttgactcc agaccagtgt 40740

cctctcctct gactccaaag ctctttatt acttcccaaa tgctgcaatg ggaaggatgc 40800

tcagaaattc tctaaccttc aaatctctct ctggacagag gaggacactc agactgcggg 40860

aagctgggac gggtcttggg tcacggaagg tggtgaccat cctgggttct taactcccag 40920

gctatgctct agccattac actgtacctc tttgtcactt agtacacagc tgctccatcc 40980

tttccctgaa caattctgtg attctaacgt cttgcctctt gaatcagagt atatgttagt 41040

aagttcttca aacgaatgaa tgatggacgg catttctttc gtagccccac tgcagaatta 41100

taccagat ctctagataa agaatagctt atgtgaaata cccctttctc actgccctgt 41160

acattccact accacctcca aatcccatca tgaacaagtc atgataggaa gcttgaaatt 41220

agaattcagc cctaaagaga ccaaaagcag aaacaaagca cacggccaca aatcagagaa 41280

ggaagggggt tttgttttgt tttttttgtt tgtttgtttt tggtcgtttg tctttctagg 41340

gccacaccccg cagcatatgg aggttcccag gctagggggtt gaatcagagc tacagctgcc 41400

ggcctatacc acagccacag caatgcggga tctgagccaa gcctacaacc tacaccacag 41460

ctcatggcaa tgctggatcc ttaacctgct gagcaaggcc agggattgaa cccacaacct 41520

tatggttcct agtcggattt atttccgctg agccatgatg ggaactccag gagttctctt 41580

ctttaagccc tttctctctc tctctctctc tctctctctc actcactcag tggttggatc 41640

atttgactcc ttttggataa tgaaagaatt agaacttggc tggtcctagc gccctgcttt 41700

gcagtgctga ttagagagag actcctataa tgtcttgytc atcacaagcc attagtgtct 41760

ggtggtgtga tttggtatca gatgctgatg ttcaccacag ccccatcgct ccttggaagg 41820

cagctatgct taccatgtac caccaacacc ccatccctcc ttggaatcaa ctctcctctg 41880

ggtcaggtgg gcccagaccc catcatacaa gcattttgaa gctgcttgct ggcttcctag 41940

gccctttctg ggagtgaccc agacaccata tggtcagctg ccgccctctc tgtgcccaaa 42000

gacaaacacg cacctcccaa cccccatctc tctcaaagca caagtctggc attttacagc 42060

tcatccagag acctattctt ggaagttccc tggtggctta gcagttaagg rtccagcgtt 42120

gtctcttgat tttctgcaga aagtgacaac ttactatttc agatgtcatt tattctactt 42180

ctggttcact ccaggctcct aggtcctaaa aataagcatc catgattttc ctaagtgtcc 42240

tccaacagga ccttgttgta cccctagatt tcctcagaat gcattctagg ccagagatct 42300

ctaggcctat aaatgtcctt cttatggcaa ccccaagcct ccctgggat tgggcctcaa 42360

ctgtggctat agaacagtgg gttgggacaa gcgtggggag attggagagt ctgggaggtc 42420

tgaagatcaa tttgttgatg cttcaattca acttaagtga tgggttcatt tccatttta 42480

gcacaactcc ttcagcaaga tatgaaatgc gtgccagcaa ccagcttgtt gaggtagaag 42540

tagtaaacat ttttagagaa actacaacag aaaatgagag ttccatattt aatgccattg 42600

accaagcaat taaaagcaac ccaagtgact ttacagaata tatccgtaag tactaaatca 42660

ttttattcaa catttacctg aagatatgtc tacctttttg tgaacttttc tattatatgt 42720

gtgaaagagg ccagaagaac tgaatcactg aatccacaga tgtctctgag tttcctttga 42780

acccgccctt tcctctctta attggcttct gctcctttat aaccattctc caagccgtca 42840

gactgactag tttgttcctc ttcttggaag gttgtcttaa taaacaacct ctggacaggg 42900

aaagatgaag aaaagagaaa caagggggct ctatgctccg ttatcacttt ccacatgctt 42960

atagaaagtt ttcctaggct gttttccttt taggcagaca cacaagtcca cagcagagct 43020

gtaacaattc attttgctgt gttaccaata ccccaaaact tagtggccca aaacattaac 43080

catctgttta ttacaatacc ataattctct gggtcaggat ttgggctggg ctcagctggg 43140

tggttctgct gatctaggca gggcttggct gatctctgct gagctcactt aagctgctgg 43200

gtgggtccac tggaggtgaa tgatctcaga tgctcacctg gatagtttgt ctctgctcca 43260

cacagtatct catcctctag caggttagtc caggtttcca aggtggcaag agggtgcgtt 43320

tcaattcaca ggtgtttctc actctccgct tgtgtcacat ttacaactgt cttcatggcc 43380

ggagcaattc acatagctga gcactgtgtg gcaggcccca ccacggggca tggatacagg 43440

aaggtaggaa aaaattgacg tgattagcac aaccaccaca gcagtcatcg ggagtttctc 43500

ttctcagagg taccttggcc agccctgggg tcggttctgt gcaactggtt atcagtcctg 43560

gttgccaaac ccaaaggcct ctgttgagac acaatttatc tgacagttct gtagcgtttg 43620

cactggtacc ctccccttcc taaagtcctc ttctctctgg ccctccatcc accattctcc 43680

tgatcccctt catcccgcct ccctcatcct tcattttgag acttccaaga ctcctgcttt 43740

accctctcac gttttgcttt ttgatgattg taacctccat gtcctcattt gcaaaacata 43800

ccactctttc taaaaataca cttaaatcct gtttgtatat agtgttcaaa catctttctc 43860

tctttctaag atgtcacaga gattctcacg gttctctcag gaacttggaa tataaatgct 43920

ctggaataaa acatgatgct gccagaaaag atggggctct cctgcaccat cccctctccg 43980

tggctgtggt ccccttagca atgtccatcc aagtatcctt tcctggcaga agagactgga 44040

gcaaaatagt tgcagagcca ttcgattttt ctcttgtcat tcatattact ccagccaggc 44100

ctgacctttc catgagcccc tgtagcatag aaaacacaca caccccttca ttcggccatt 44160

tttcttttct ttctcacttt tcatgtttaa ctctttgctc attgccatca gattttcttc 44220

ttcttgttat tgttttcggt tttgtttgtg gcaaacctat gctcatttgg actctagacg 44280

gtctctgtca ttcttttaat attccatctt tggttaagag accttctttg catattctgt 44340

ctatatctct cacctgagag tttcctgtgc gaaccatgcc agtttcttaa attaactcac 44400

cctctcccat ggtttttttg tagttcattg ggaccatatg cagctgcaca gtgagaatta 44460

cattttcaga gcttcttact gttttaacct attttcatct tcctgagtct catgccatgg 44520

aactgtacct taccctttct gaacacttca aaatgtatct ctcaaagtca tacttctcta 44580

tctggctatc acttggtcct tatcttcaaa aggaccccatc gcttctgtct gccaatatat 44640

ttcttcttgg ggtcataatt tgctccagag tagtaattca tgtagtcaga acctctgccc 44700

cctggaagat gaaactgtca atgaggcaga caaaacctta tcatacgctc cgtttggggc 44760

tggatgagac ttctaacaag tgtgaggaag ttgagtgtcc ccgtcagcat ggtcaccaga 44820

tgaaaacaac tggtgcattc ttcaagavaa agttggggtt ccctttgtaa ttctctcatt 44880

tgactctact tagtaaccac cttggctctg agctttggca gagttgaaac catgtggcat 44940

cagcactgag aagtgctctt tttttttttt tttgatatct cctcttaaca cgtctctact 45000

ctgaggccag gcatgtgctc tcagatacat taacaaattc ctgaatgatg ccaactagag 45060

gtggccacaa gagcatgaat cacccagcaa atagaagtga gagcttagga cttttagaga 45120

aaaacacctt ttgctgctct cttgtttctg ctgcaaatac ggactctctc ctttgtgtca 45180

aaagctgtag aatctctttc tctaggtttt tacttggagg catttttaaa tggcaggtaa 45240

tgaaggaagg ttagttacag ctttgggatg tttagaagag agcaaccaca tctctgttct 45300

acctgctcct gctctctgct cagattcatc ttttgtccct aatgccttcc attatgcttc 45360

tacactattt ttaaaagatg gctttctmgt gccatgtttc tgctcagaat ccaccaattg 45420

attcttaggc ttacaggata aaagtcagtg ctcttagcct ggacccaaac cctgctatac 45480

caacgaatga acaagagaat gattatttgt tgagtttcct gtagtggaca cttgcattta 45540

tatgagctca ttcagttcta acartaatga tgtgaaacaa gaatcattag ccccatttca 45600

tagagggagg gaacttggac ctaccacaaa tcaaataatt gaacaaaaag cccacagcca 45660

atggggacgg caaagccaag aagtctcagt gttcatcccc ctcattgcat tgcctctccc 45720

ctgcccaagg acaactccat cactcctgga caggaatcct ttacttcttt ccataccttg 45780

tttattccac ataggcagac tctgcctccc ctttgactgt cacatcttcc ttatctttca 45840

agacttagcc caattcctgc ctcctccaca atgttacctt taggtagctg agcccctgaa 45900

ttcccattgt acttaaagtc tgtgttacaa gttgctggat gcaactggct atgggaacca 45960

acacgtaact ctccaaagat ggggcccaaa acttaaccaa gtactgtgtc ccttgtggaa 46020

cctagtcaag agctaggttc agaagagatc ttgaacttga tcgtaattgc gttgcactgg 46080

attgaattca tttgaattgg ataatgtaag gaaattgttc tagaagagat gattggggag 46140

ttgagtttca aaagcccaga tttctattgg ttgaccccca agacatgtta aaaggcaacc 46200

gtaacatcag tggaatgggg tgctttcaca agctaccctg tagtgtcttg ctaaatacgt 46260

ctttgaattc tttgtgcctc tcagagcaag atcggtgtga ttattatgga tgtaagaggc 46320

aaaatgagca agatgagtgc cccagtggtt tactgtgtga gtgcaaagag gggctggaga 46380

gaccaaaccc acagatccct ctgtgtgttg gtaagtgttc cacctgcttc tgaatggtcc 46440

tcacacctcc tgccttttct ccattcacat ttgcaaagga ctcattttct ggctcaagtt 46500

tccatgtaca tttcagagtc tgagggatgt gaattttggc tataggcctg tggtcctgga 46560

tggggcgtgg gctaaggagc tggtgaggag gcccatgtgc tgcctcacag gggacaagtc 46620

acatgcagag ccatggagcg cctcattcta cccagaaaaa gcaatgtgaa aaaattgtat 46680

tctgtgacat gtcattttgg ggttgaacgg gttgaactgg tcgtttctga aaattaccca 46740

gaaatcgctc tgttttgaag ctttgggtcc agcatgctct gatacctgca atgcagagaa 46800

caataggcaa tgcctcgtga ggagcgctgg gaacgccgaa tgcttgtgcc tgcctggcta 46860

caaggaggat aaaaacaaga tctgccaagc gtaagagaaa tcactccttt ttcatcttgt 46920

catagatgaa gctgagagag aagttagcac gtgtcacata catatgggtt tagggtcagg 46980

aagctggtac gctggcatcc gttaggaagt tgcaaagcgt caccaccttc atcatcacca 47040

ttcccatcac catcgggttt tgaaccttct ctatgtgctg ggaaccaccc aagcaatact 47100

tgtcacacag cagcactgtc gggcctgtgt tcgttccctg tcttcaggtg aggaagctga 47160

agttcggaga atataaacat tccccagatca cagggcagtg aataagtagc agagccatgt 47220

ggcctcacag gctgaggggg tcccacaatg ctccatgcct ctggggctct gctccgccca 47280

aactgctgaa tgttcctgat gctcggaggg yggagcgttc agtcagtagc aattggaacc 47340

acaggagatt ggggagcacc tccctcaacc ttccggaggc cgtgaaaggt actcccctga 47400

agaatgggag tgtaaggaag aaaagagaaa gaggagatgg aacagggagg agagagtgaa 47460

agaaagatgc aaggatggca agttcccatt gtggcacagt ggaaacaatc caactaatat 47520

gcatgagaat gcgggttcca tccctggcct tgctcagtgg tttaaggatc tggcattgcc 47580

cggagctgtg gtgtaggtcg cagactcagc tcagatcttg agtggctgtg gctgtggctg 47640

tggttgtggc tgtggcatag gctggcagct gtagccctgg tttgactcct agcctgggaa 47700

tttccatatg ctacaagtgc agccgtaaaa aaaaaaaaaa aaaaaagaaa agaaaaatgc 47760

caggatgaca tggaggttgg gtgggacctc tgatgtgctc ctccaaggct ctttaccatg 47820

gtttccttct cattgcattt taggtgtcca tttggctaca gtggagttgg ctgtgaaaac 47880

cgtgagtaga aaagtcttgc tctgttattc agcttctgaa aaccatggac cacattgaga 47940

attctaccca gtcccccaaa ggccctggct tgtttcataa agagagagaa aatctgcctg 48000

tagtttgcct tctgggtcag ccatcccatg tccaccctta ggtcccagac agagcccctt 48060

cagtgtagga aataacagaa gtccacccaa ggggtgcact ggtgcctctg gacttgtctc 48120

aggccaaaga agtttgtccc aacattatat ttgtttccta cgttagcttc aatctgcatt 48180

gtctgccact ggaaaggata caagactrta ttccaaaact gtgttcatgg gcgagcagaa 48240

ctagaaactg gacttttaac tgctcaagtt ttctttttgt ctcagagctc tttgctagtt 48300

tgggatgtgt tattgcaaat tcagttgctt tgttccctcc tgccagattg ggacaatact 48360

aacttcatat tttacattat tctttttttt tttttttrtc tttgtccttt ttagggctgc 48420

accctcggca tatggaggtt cccaggctag gggtctaatc agagctgtac ccaccggcct 48480

aagccacagc cacagcaaca ccagatctga gctgcgtctg cgacctacac cacagctcac 48540

ggcaacgccg gatccttaat ccgctgagcg agaccaggga ttgaacctgc aacctcctgg 48600

ttcctagtcg gatttgtkrr staaccactg agccacgaca ggcactcctt aaattattct 48660

atatttaaaa ttgtacgtaa cgcaccaaaa gatcaatgac ccaaaatgaa cgaaatattc 48720

ctattcacct tccagccagc cacaaagtta gtgatgagaa atgytttcag tttccgctgt 48780

attttttttt tttctttttt ggccgcccca cagcgtatgg agttccctgg gccaaggatc 48840

agatccaagc tgcagtatcc acttacgctg cagctgcagc aatgctggat ccttaaccct 48900

cctgcatccc agtgctcccg acacactacc tctcccttcr cgccacttca ggaactccaa 48960

tttccactgt attttgtttc aaggctgggt ttgctaagtc atttcttcct tctggctttc 49020

agtgtccaac ctataaagtg agaggtttgg atcagataac rtttgaagtt tttccagacg 49080

gaacagtctg tggcccatga accccaggtta ggggtggcat acgtggcagg catgtagcag 49140

aagaatcaag gggatctaga gtcgtgctgt ccaatagaaa tatgacgtga acttcagatt 49200

tcatttcaca tttaaaagtt ctaagcaaaa ttttaaaggt acagagaaat agatgaaatt 49260

gatttttaac attttgtttc attcartaca ttgaaaatag tatcatttca acaggtaatc 49320

aatataagaa tattaataag gggagttccc gtcatggcgc agtggttaac aaatctgact 49380

acggacyatg gggttgaaga ttcgatccct ggccttgctc agtgggttaa ggatccggtg 49440

ttgccgtgag ctgtggtata ggtcgcagac gcggcttgga tcccaagttg ctgtggctct 49500

ggcgcaggct ggcagctaca gctccaatta aacccctagc ctgggaacct ccatatgctg 49560

tgggaacgrc cctagaaaag gcaaaaagac caaaaaaaaa aaaaaagaag agaatattaa 49620

taagatattt tacatttttt tcatactatg tgtttggaat cactgcgtat gttacgcttc 49680

caggccatcc cacagccaca tttcgagtgc tcagtaacat tagagcagat ctagaggaaa 49740

aagggtggta atgaacctcc agacttttcc ctccccaatt ccatctggac atcacccggc 49800

tcctctatca gggacaaatt taactcccag ccaggtcccc tggtttatcg gcaaggtcag 49860

ttgtggcagg cargtgagag cagcagaagc agggagatgg catggacctc ggaygagatc 49920

ccatccccat gaagcccagg aaggtcctgc caccctggcc tactttgtgt ttaggtcctt 49980

gattatctgg caaggctgcc tcttctgaac acacagttgg gggtgagcag acaggggtt 50040

agagaaggtg agccaccac ctatcctggc cctggagacc agagcctgat ggtcctcagg 50100

gccaggagcc actaggaacg cgtgtacctg tgcagtagac agtggaggtc tgcttggagg 50160

aggctgttcc tactcttctc ttctcccttc cagcatatca gctgatcctc accatcgtgg 50220

gcaccatcgc tggcatcctc attctcagcc tggtgattgc attcgtcctc tcattaaggt 50280

atggaatggt gtctctttcc tgggacacag cgycactggc ctcacactct ggttcgcaaa 50340

cacacccaag ccttttccca ggcaagagga gaaaaataag gcgaggcctc tctagcacca 50400

ttgccctttt ttcccgacct taattttctc ctgattctat aagctacaca tagtccctga 50460

agaaaayagg gaaacgtgca aagaaacaga ataaaaaatc accccacaaaa tccactaccc 50520

tgaagcaacc aacattttgg catatatctt tccaggtttt ttcctacctt ttaaaacaat 50580

attgtatatg taactttttt tttcawtttg tgcctttttt tttcattttt tttaagtttt 50640

attgaggtag agtggattta caatgttgtg gtgatttctg ctttacaaat aaagtgattc 50700

tgttatgcat gtacccacat ccattctctt tcagattctt ttcccacatg gattatcaca 50760

aaatattggg tcaagttccc tgtgctatac agcaggttcc tgttggccaa tcattctata 50820

tacctcagtg agcaaaggcc aatcccaaac cccccctctg ttccattccc tcccgcctcc 50880

ccctccccct cagcaaccac aagtctgttc tctaggtctg tgagtctgtt tctgtttttt 50940

gcagataggt tcatttgtgc catattttag attccacgta taagtgatat catatctgta 51000

tacatagttt ttatcttttg cgtttgtaaa atttgttgtc gaggtttgat tttgcttaat 51060

actgcaatgt gatgtagcag tcagttttct gatgtcatta aaaaactcta gttagcggta 51120

tttaactcca ctgatgttgt caaatgtaat ttacaagtca ctcccctgct caagtccctc 51180

caaagactct aacaaccacc tcaactgagc gccctgggct ttaagtagaa atgttatctc 51240

cgtgtggcca tttctcagct tgatgcctta aatgctctct ttaaaaactt aagtaggggtc 51300

cattcattta ttcagcaaat atttcgatat aataaaattc cctccattaa gagggatgct 51360

atcatctatt attatctatt atttattcta ttctaaatgt gccaagtgag gatctacttt 51420

gaccaaaaaa aaaaaaaaaa aagctacttt tggaattccc gtcatggcac agcatgttac 51480

caaatccgac taggaaccat gaggttgcag gttcaatccc tggccttgct cattgggtta 51540

aggatctcgc attgctgtgg ctgtggtgta ggccagcagc tacagccctg attagacccc 51600

tagcctgaga acctccatat gccatgggag cagccctaga aaaggcaaaa agacaaaaaa 51660

aaaaaaaaaa aagctacttt taaaaaataa cattgagcac atatcttata ttttagctaa 51720

ataggttttt ttttcactat aatatggaca aatagatttt aggttcctaa atatttgact 51780

tcctcttctt ttttttctct ttaatccatt ttgatatgaa ctctccccag agacttgatt 51840

acaagggcac cacactggga actggaaagg caactttccc tttccagtca gagggccagg 51900

atgccaggct ctggagcagg tctgccctct tggcaggagg actgactcac tgccctggtc 51960

tccagggtcc aggggctgca gcagaacttg caagcccgtg acagccacag ttttcagtcc 52020

ccccttcctg ctgcagagcc aggtttgctc ttaattcagc cttagggtca tgattcttgg 52080

gagcacaccc rcaattgtca tttcttgccc atttcaaagg gaaggaagcc tctcctcagt 52140

cacmctagat gggagtgttg tcacagcarg aaccctataa tgggctggrc tgaccatcag 52200

gtcaccgggt gaggggrtct tgacgacatt ccagtgtctg agagacacct gagamtactc 52260

tgtggccatc acccatgcct gtggccccac tgcccactct ccagggccag gaccatacct 52320

gggggccaga ggacagaaag aagggttaga ggatcagagt tacttggggg cagatgccty 52380

cctcacctgc aagtccttcy acttggcttt gaayrcttac wcttgagggc ccatagcaga 52440

acctgggaaa ctcctattty tccttttaag ayctttatwt cttgaattca ttttattcak 52500

ggagskgytt ctgtagatac tgactttgcc mraatgaaaa atwygtcaca awtatyttct 52560

ttctttgact tttattatca tttccataag taattcaaag gcatacttaa raatactrac 52620

aatattagca aaatataaac aggactggtg gaattagagg gttttagttt tgatggtttc 52680

atggagtttc tcaggagagt tagggaatct acactgaatg aagcaatgaa gtagaaccgg 52740

taatgggcat caggtctgca acttactctt aaatgrttca aaaatgttaa cagttgggga 52800

atctgggtga gagatgtatg ggagctctat gtaatttgca tgtgtgtgac ttacgtgtaa 52860

atttgaaatt atttcaacat ttttaaacat tttgaaaaaa gccttattaac atcagtttca 52920

agggacttgc tgccattgtc agatcctaat tcctgtgact gattacagat caaacaacaa 52980

aaacaagaat gtggaagaac agaacttgat tgagaaygac tttcaaaacc tgaaactgca 53040

ggagactggc tttagcaacc taggagcagg aaacctcttt cctaagatca aggtggacat 53100

ttccagaggt ggtcagccac agaatcccta tataacccmg ggtagtatac cccgccctga 53160

ctattaggtg agtctttggg tgctttctgt ttgcttttgc tgagtgaaat gccccagagc 53220

agacttggaa cctgcctagt gtatcatcac cagccctggc tgccctgtgg gtttgggttt 53280

tgggttttkg ggggtttgtt tttttgaatc aaaagttact ggagttyccg tcctggctca 53340

gtggttaaca aatccgacta ggaaccatga ggttgcgggt ttgatccctg gcctcactca 53400

gtggggtaag gctccggcat tgccgtgagc tgtggtgtag gttgcatact cggcttggat 53460

cccacgttgc tgtggctctg gcgtaggccg tcggctacag ctctgattag accccttgtc 53520

tgagaaccgg ctacagctyt gattagaccc cttgtytgag aacctccata tgctgtgggt 53580

gcagctctwg aaaagacaaa aaaagacaaa agaaaaaaaa aaaaagttwt tgtaatascc 53640

mccgcttgat ccgtagcact agccatctct gtgactttar acacacaact taatttctct 53700

gaccatcgat ttcctcagct gtaaaaaggg aagcttacat taracaccct taaratctct 53760

ttacatcctc tgttcctatg gctgtaacgc tggkggttgt gatgacaatt ccaaractgc 53820

tgaagggccc aggacctagg ccagaaatgg gaaggactgt gggatgtaaa cacagttttg 53880

ttcacgtggw tcctttccag ccascccttc taaagraaaa tctctccttt cttycctccc 53940

tcagacctca aggcaactat atcaatttct aaaagaacaa gacaagtgag gctggtaata 54000

ctttctttcc cctggaggam tttcaaggag catagcttgt cccgttagat ttgtcttgga 54060

cagtcatggc cagcaatgct gtatgacccg atggaatgct gaaagcagaa agcccatcag 54120

agagcactcc tgaaggaaag cgtgacaaga tgaaggcggg agagatggca aaggaggagg 54180

aggggccaga gaggagagggg gggaaggaag agaggcgagg agagcagcga cttgtttgaa 54240

aaaggaaatg ctggcattcc agtcgtggcg cagtggttaa cgaatccact aggaaccatg 54300

aggttgttgg ttcgatccct gtcctcgctc aatgggttaa ggatctggca ttgccgtgag 54360

ctgtggtgta gctcgcagac gtagctcgga tcccgtgttk ctgtggctgt ggtgtgtagg 54420

cctgcagctg tagctccaat tagaccccta gcctggaaac ttctgtatgc catgggtgca 54480

gccctaaaag gacaaaagaa aaaaaagaaa aagaaaaaaa aaggaaatat tcagagaaaa 54540

ggaaagaagg agggccagga gtcaggcatt cccaggagag aagaaaaata aagagtctga 54600

aagaggaggg gacccagaag ggtggtagag aggctgggga agccccccta ccccggcccc 54660

caacatccac actctctgtg tgcatatttc acttccatgg gctgccctca gcagatggat 54720

cgtcctttga ggatcttaat gggaaaggct gacttttatt ttgaaatccc agggttgatt 54780

ttagacattg tttcctttag tcatgaaatc taagtgttac atgtgtgcac aaagggatga 54840

cttgtaccac ctgggccatg tggggtcaga cacagagtct ggtagcagga gctgagagtg 54900

gatgtcctgt tgggtctacc tgaggtgaac cttaaccatc aaactcattt cagacttccc 54960

ttctccacat cagtggagcc cgtacttcag gtataacctc tgaagaaagc ccttgagtgg 55020

ctttcccatg cttataggat ttagaattgg acagacctag gttcatagcc aggctcttcc 55080

tgccactagc tgtgtgatct ccaatgagtt gtttgacttc tgtgggtttg ttttctcatc 55140

tgtgaaacgg acaaaatgaa acctacctca caggataaag atcaaataag caaatatgca 55200

aaaagaactc agcaccatga tgggttctaa tttaaataac agtattcaaa taatggtaat 55260

taaaataatg ttaatcattt atgaatttaa attttctcat aggcttctgt tggaatgttt 55320

ctaagagttt ctctctctct ttctctcttt ttgtctagaa tggtaaaaat ttggaaccat 55380

ccatggccgt caacccagca catgagattt ttgtgtgagg gtttgataga ctgatggaga 55440

aaagagatct ggcctttagt atttttcctc catccaacat ctggtctctc tgaatggaag 55500

ctgtaactgt ttgcattgaa ttcagcgtgt ttgaatctaa gtacctatga agttaaatgc 55560

acacataaat gctattagca cttgtaagat agttgatttg ctccagtaca gtggttaggt 55620

gtttgtttgt tttttcctgg tgggacaaca ataaccattt ccaaactaga gggaagctcc 55680

ccaggattgt tcattcctgg aagagtctaa gtgttgcttt gagttgatag catgcagata 55740

agtgacctaa ttctcggaaa ttcttacaca tcagctgtgg aattccagta gacgggagga 55800

gacaccatcc ttaaaggtac tcagtggacc ctgaagattc attctctagc aagttaaaat 55860

aaggatgcta cttctggtcc tcaaatgtgg gaatttcatt acccttaaac tgcttgcgac 55920

ctggcttcac accaagagga cagaaaacaa gacatgtctc ccagctttca ggaaggagtt 55980

cctgggaatc caggaccttt gagatgttaa ttaaaagatg tggacttggg agttcccgtc 56040

gtggcgcagt ggttaacgaa tccgactagg aaccatgagg ttgcgggttc ggtccctgcc 56100

cttgctcagt gggttaatga tccggcgttg ccgtgagctg tggtgtaggt tgcagacgcg 56160

gctcggatcc tgcgttgctg tggatctggc gtaggccgga ggctacagct ccgattcgac 56220

ccctagcatg ggaacctcca tatgccgcgg gagtggccca agaaatagca aaaaaaaaaa 56280

aaaaaaaaaa aaaaaaaaaa gatgtggact tgagatgtct tccatgagga tacattctag 56340

agtctttgaa atgctggtgg gtgagaccaa gagtagagc aacagaggca gaaggaagct 56400

gagctgggag gaagaaggta cccaagaccc accttcatcc tcagattccc ctccccaata 56460

aacaggtggg cactttctct aatgcatgcc ctacctatga ctcaagacct atgaaaggat 56520

gtgaagataa aaaccctgga gcaagagttt gagtcactt tgaaattgta gacattctgt 56580

ctggctgatg gtggagaagt gatgcagatt ccagaggtac ttcaccaaag tctcagcctt 56640

ccctggggac agaggcttag cagtggccac caacacacac acgcacatac acagccaaga 56700

atggggcagc cacctgccgc acaagctt 56728

SEQ ID NO3

<210>3

<211>1796

<212>PRT

<400>3

MKVFIHFTLL ALLFLKSGKE HLHIFNSFRH ISSAIIYTVF CLLKLCLSAS MAQNKTGGTT 60

KLTSTENTSS PNAAESTSAP SAARNSAPGT AASTSAPSAA ASTSAPAAGA STSAPAAGAS 120

TSAPSVAAST SAPAAGASTS APSAASTSAP AAGASTSAPA AAASTSAPAP AASTSAAAAS 180

STSAPAVAAS TSAPAPAAST SAPSAAASTS APAAGASTSA PDPTASTSAQ AAEASTSAPS 240

AAASTSAPAA GASTSAPSVA ASTSTPAAGA STSGPAATAS TSAPATGAST SAPSVAASTS 300

TPAAGASTSG PAAAASTSAP AAGASTSAPS VAASTSTPAA GASTSGPAAA ASTSAPATGA 360

STSGPAAAAS TSAPAVGAST SGPAASASTS ALSEAESTSS PAAGASTSAP SVAASTSAPA 420

AAASTSAPSA ASTSSPDPTA STSAPAAGAS TSAPSAAAST SAPAAGASTS APSAAASTSS 480

PAAGASTSAP SAASTSAPSV AASTSAPAPV ASTSAPAPGA STSAPSAAST SAPATGASTS 540

APSAAASTSA PSTAASTSAP AAGASTSAPA PAASTSAPAP AASTSAPAPA ASTSAPAIGA 600

STSAPSAAST SAPSAASTSA PAAGASTSAPA SVAASTSAPA PAASTSAPAA GASTSAPSAA 660

STSAPAAGAS TSAPAAAAST STPDPTASTS APAAGASTSA PAPAASSPAP AAGTSTSGPA 720

PAASTSAPAA AASTSPLDPT ASTSTPAAGA STSAPSAAAS TSAPSAASTS APSAASTSAP 780

AAGASTSAPA VEASTSAPAP AASTSAPAAG ASTSAPSAAS TSAPAAGAST SAPAAAASTS 840

APAPTASTSA PAAVASTSAA APAATTSALA AVASTNPPAA GASTSSPAAG ASTSAPSVAA 900

STSAPAPVAS TSAPAAGAST SAPSAASTSA PAAGASTSAP APGASTSAPA PAASTSAPSA 960

ASTSAPAIGA STSGPAPAAS TSAPAAAAST SAPPDPTASTS TPAAGASTSA PSAAASTSAP 1020

APAASTSAPS AASTSALDPT ASTSAPSAAS TSAPAAGAST SAPAPAASTS TPAAGASTSA 1080

PSAASTSAPA AGASTSAPAP TASTSAPAAV ASTSAAAPTA STSAPAAAAS TSTLAPTAST 1140

SAPAPAASTS APSAASTSAP AAAASTSAPA PAASTSAPAG AASTSAPAAG ASTSAPSAAS 1200

TSAPAAAAST SAPAAGASTS APAAAASTSA PAPAASTSAP AAVASTSAPA PGASTSAPSV 1260

ASTSAPAPAA STSSPAAGAS TSAPSAASTS APVPAASTSS PAAGASTSAP APAASTSSPA 1320

AGASTSAPAA GASTSAPAAG ASTSAPAPAA STSSPAAGAS TSAPAPAAST SAAAAGASTS 1380

APSAESTSAP AAGASTSAPA AGASTSAPAA GASTSAPSAA STSAPAAAAS TTPASAANAS 1440

SAISSTPIAS TPTSGSRNPC QQDSCKGGSS CVSLNNTFFC LCSEGYYYNS STCNKGKIFP 1500

GKITVKISET TDLEHENSVT YQNLYLQVTN FFNDTFSNSE YGQTVILKVS TTPSARYEMR 1560

ASNQLVEVEV VNIFRETTTE NESSIFNAID QAIKSNPSDF TEYIQQDRCD YYGCKRQNEQ 1620

DECPSGLLCE CKEGLERPNP QIPLCVALGP ACSDTCNAEN NRQCLVRSAG NAECLCLPGY 1680

KEDKNKICQA CPFGYSGVGC ENPYQLILTI VGTIAGILIL SLVIAFVLSL RSNNKNKNVE 1740

EQNLIENDFQ NLKLQETGFS NLGAGNLFPK IKVDISRGGQ PQNPYITXGS IPRPDY 1796

Claims (7)

1. A MUC13 gene that determines susceptibility/resistance to piglet F4ac diarrhea, characterized in that: its cDNA sequence features are as shown in the sequence table SEQ ID NO 1. 2. The MUC13 gene that determines piglet F4ac diarrhea susceptibility/resistance as claimed in claim 1, characterized in that: its DNA sequence is characterized by the sequence table SEQ ID NO 2. 3. The MUC13 gene that determines piglet F4ac diarrhea susceptibility/resistance as claimed in claim 1, is characterized in that: its encoded protein amino acid sequence is as sequence table SEQ ID NO 3. 4. the MUC13 gene of determining piglet F4ac diarrhea susceptibility/resistance as claimed in claim 2, is characterized in that: the 32813bp place of its sequence table SEQ ID NO 2 is the base mutation of C32813-32813T; The 37618bp place is The base mutation of C37618-37618T; the base mutation of A42785-42785C at 42785bp; the base mutation of A46531-46531C at 46531bp; the base mutation of A53019-53019C at 53019bp. 5. The MUC13 gene for determining piglet F4ac diarrhea susceptibility/resistance according to claim 2, characterized in that: the primer sequence used to isolate the MUC13 gene is: Isolate MUC13 Gene Primer Information 6. The MUC13 gene for determining piglet F4ac diarrhea susceptibility/resistance according to claim 4, characterized in that: the base mutation primer sequence for detecting the MUC13 gene is: Primer information for determining 5 key SNP markers of MUC13 gene

7. The application of the MUC13 gene and its SNP site, which determines the susceptibility/resistance to piglet F4ac diarrhea, in the selection and breeding of pigs to improve the anti-diarrheal disease traits.

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