CN101372712A - 一个可鉴别西方猪种断奶仔猪f4腹泻易感性/抗性的muc4分子标记及应用 - Google Patents
一个可鉴别西方猪种断奶仔猪f4腹泻易感性/抗性的muc4分子标记及应用 Download PDFInfo
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Abstract
本发明公开了一个可鉴别西方猪种断奶仔猪F4腹泻易感性/抗性、可用于种猪遗传改良的MUC4分子标记及其MUC4分子标记在选育改良种猪抗腹泻病性状中的应用，它先进行MUC4多态性位点g.13383C＞T，g.15580A＞G，g.21208A＞G和g.26002A＞C的鉴别，再采用PCR－RFLP方法对每个多态位点进行基因型判定，然后构建单倍型。本发明通过现代分子生物学技术检测这4个多态位点，构建单倍型，利用标记辅助选择(MAS)选择有利的单倍型对个体留种，可以显著提高种群断奶仔猪的抗腹泻病能力，大大减少仔猪死亡率。
Description
技术领域
本发明涉及一个可鉴别西方猪种断奶仔猪F4腹泻易感性/抗性、可用于种猪遗传改良的MUC4分子标记及其MUC4分子标记在选育改良种猪抗腹泻病性状中的应用。
背景技术
仔猪腹泻是养猪生产中的常见传染病，给世界养猪业造成了巨大经济损失。丹麦养猪业大规模的场间调查表明:6％-7％的新生仔猪患有腹泻病，使断奶前仔猪的死亡率达2.7％，占此期间整个死亡率的11.9％。即使是病愈个体，也会严重影响生长发育，有的形成僵猪，导致饲养成本增加，销售收入下降。
产肠毒素大肠杆菌(Enterotoxigenic Escherichia coli，ETEC)F4是引发新生和断奶前仔猪腹泻病的最主要致病菌，以我国和丹麦为例，分别有约35％和40％的仔猪腹泻病是因ETEC F4侵染而引起的。在我国，其致死率在10％---30％之间，在个别地方甚至比这个比例更高。
ETEC F4根据免疫学特征分为ab，ac和ad三种亚型，其中F4ac是最主要的致病原，在美国、西班牙和韩国，ETEC仔猪腹泻病例中只分离到F4ac亚型。ETEC F4入侵猪肠道后，通过其特异性黏附素附着于小肠上皮细胞表面的受体，大量繁殖产生肠毒素，致使肠上皮细胞分泌功能亢进，大量水和电解质进入肠腔，造成肠腔中大量液体积蓄，超过了肠壁重吸收能力而引起腹泻。由此可见，仔猪小肠上皮细胞有无ETEC F4受体，特别是有无F4ac受体是仔猪被ETEC F4感染时是否发病的关键。无受体猪表现为抵抗型，有受体猪则表现为易感型。分离和鉴别猪ETEC F4ac受体基因及其抗性位点因而成为猪ETEC F4ac腹泻抗病育种的关键。
Grange等对F4受体的性质做了一系列详细的研究，在猪小肠上皮细胞分离到两种粘液型唾液酸糖蛋白(Intestine mucus glycoprotein，IMTGP)，分子量分别为210kDa(IMTGP-1)和240kDa(IMTGP-2)，这两种蛋白被认为是猪对F4ac易感性和抗性重要的决定因子。瑞典/丹麦的Anderson研究小组及瑞士Vogeli研究小组先后将ETEC F4ac受体基因定位于猪13号染色体q41区域。最近，Joller等通过精细定位进一步将ETEC F4ac受体基因定位于q41区域SW207与S0283之间。申请人通过利用大规模白色杜洛克×二花脸资源家系群体在13号染色体上采用高密度微卫星标记将F4ac受体基因精细定位于S0283邻近区域。MUC4基因位于该区域，结合其生理生化功能分析，认为MUC4基因极有可能是肠毒素大肠杆菌F4ac受体的一个重要候选基因。
鉴于以上背景情况，申请人深入开展了猪MUC4基因多态性的搜寻、鉴别及其影响仔猪ETEC F4腹泻病性状的研究，以期建立标记辅助选择技术开展断奶仔猪腹泻病的抗病育种工作。
发明内容
本发明的第一个目的是提供一个可鉴别西方猪种断奶仔猪F4腹泻易感性/抗性的MUC4分子标记，通过现代分子生物学技术检测该标记，利用该标记辅助选择(MAS)选择有利的个体留种，以显著提高种群断奶仔猪的抗腹泻病能力，大大降低仔猪死亡率。
本发明的第二个目的是MUC4分子标记在选育改良种猪抗腹泻病性状中的应用。
本发明的第一个目的是这样实现的:
(1)多态性位点(SNP)的鉴别:
依据猪的MUC4基因部分全基因组序列(DQ848681)，大致每隔6kb左右用公用的Primer 5.0或Oligo 6.0软件设计一对引物开发一个特异性标签序列(STS)，共设计4对引物:STS13383 FP/RP；STS15580 FP/RP；STS21208FP/RP和STS26002 FP/RP(详见表1)。扩增猪基因组DNA(脱氧核糖核酸)。
表1 MUC4特异性STS开发与多态位点基因分型引物
注：以上引物设计主要参照猪MUC4基因序列(GenBank收录号为DQ848681)
25μL的聚合酶链式反应(PCR)反应体系中，包括40ng猪基因组DNA，1.0mM MgCl2，100mM dNTP，正反向引物各10pmol，3单位DNA聚合酶(Taq酶)及1×PCR buffer(缓冲液)(上海博采公司)；扩增条件为:95℃3min；94℃ 60s，53-63℃ 30s，72℃ 45-90s，共35个循环；最后在72℃延伸10min；PCR扩增产物采用QIAquick DNA纯化试剂盒(QIAGEN，Hilden，德国)纯化合后委托上海生工生物工程有限公司直接测序，测序结果利用BLAST Search(http://w.ncbi.nlm.nih.gov/BLAST1)在线比对验证，然后使用公用的DNAStar的SeqMan软件分析测序结果，发现4个STS上分别存在一个多态位点(SNP):g.13383C>T，g.15580A>G，g.21208A>G和g.26002A>C；
(2)位于猪MUC4序列的第13383、15580、21208和26002等4个核苷酸位点为:
1 gctactcatg gtacttctga ttggggttcc tctgataatc aggatgttga ccatatattc
61 atgagctggt tgtccatctg tatctctttc ttgaggaaat gtctattcag gtcctttgcc
121 catttttcac ttgggttgtt ggctgttgag ttgtaaaagt tgcttatata ttctagagat
181 taagccctta tcagctgcat catttcaaac tattttctcc cattctgtaa gttgcctttt
241 tgtttatttt tggtttcctg tgctgtgcaa aagcttgtca gttcgattag gtgtcattgg
301 tttacttttg cttttatttc tgttgctttc ggaggctgac ctgagaaaac atttgtaagg
361 ttgatgtcag agaatgtttt gcctatgtta tcttccagga gtttgattgt gtcttgtctt
421 atacttaact ctttaagcca ttttgagttt attttcatga tttgcatgca gctgtccaca
481 tttcccagca attcctgctt aataaaaatt tgtgtatgta ttgcttgaaa ggacagtctg
541 tgtattttaa ttaaaaaact gttttgagac tgaagctctt gtatcttgcc ttccatatat
601 ggcttatatg gagagagtag gagtgttctg ggtccatttc cattatacca cacaatgaat
661 gacactgaaa acatattagc acaaagatat tcactgaagt attatttatg atgaggttac
721 acagtggccg tagaaatatt agacaggggt aatcctgcag ccacttattc tgacccacca
781 tgatatacag tggtacacgc gggaaaaaat tgtgttcttt tttttgtttg ttttgttttg
841 tttttttttt gtttgtttgt ttgtttttgt ctntttgcca tttcttgggc cactcccatg
901 gcatatggag gttcccgggc taggggtcta accggagctg tagctgccag cctacaccag
961 agccacagca acgcgggatc caagccgagt ctgcaaccta caccacagct cacggcaacg
1021 ccagatcctt aacccactga gcaaggccag gcatcgaacc cgcaacctca tgcttcctag
1081 tcggattcgt taagcactgc gccacgatgg gaactccaaa atactgtttt ttttgttttg
1141 ttttgtttat ttgtttttgt ttttgttttt ttccttttct agggccgctc cctcggcata
1201 tggagtttcc caggttagaa gtcaaattgg agctgcagct gccggcctac accagagcca
1261 cagcaacatg ggatctgagc cacatctacg acctacacca cagctcacgg caacgccgga
1321 tccttaaccc actgagcgag gccagggatc gaacccacaa cctcatggtt cctagtcagg
1381 ttcattaacc actgtgccac agtgggaact ccaaaattgt gttcttaatg ctacaacttc
1441 acagaaggaa cctgtcttgt caacgagtcc ttgtatctta tatttaacgg cactattgaa
1501 acatatgcat catcatatct ttaatcctct acaggtggaa acactatgac aactactatt
1561 actgaggaga aatacccatc gaatatcagc tacataactc ccacaaaaga aaacctctca
1621 aaagaagtaa catcaacaaa caaaactgaa agaaaaaaga gggaaacttc aaattctggt
1681 acttacaaat caactatggt gagttcatca gtctcagcga caactggacc agggccccag
1741 tcaacctcac cctcttccac cacctctgct gaggggacag cagcatcttc tctggtccac
1801 cagactcaga gcatggagac caccagagaa acccacagca gcgccatcgc agctgtgagc
1861 tcctcgacac ctttatcttc cacaagtgga caccctctta cagaaggcgc ttcccgggag
1921 acatcccctt cgggggaccc aacctcttca tccgcgtcca gacccacccc cacacctgca
1981 acaacatcag ccgtgtcgac agcacctgct tctacagatg gcacttcccc cacttccagc
2041 ctaagcaaca cacccccaac aacatcacgg gtggtcacatccccagttac aaccgacacc
2101 actgtgggag gcccagggga cacgtcccca cccggcacaa gaaccatcac tccaggcacc
2161 tcatcagtct caacgacaac tggaccaggg ccccagtcaa cctcaccctc ttccaccacc
2221 tctgctgagg ggacagcagc atcttctctg gtccaccaga ctcagagcat ggagaccacc
2281 agagaaaccc acagcagcgc catcgcagct gtgagctcct cgacaccttt atcttccaca
2341 agtggacacc ctcttacaga aggcgcttcc cgggagacat ccccttcagg tgaaacaagc
2401 tcttcagcca catttagacc gacccccata tctgtgtcgt gtttccatac ataaaccagt
2461 atttcttctt tttctgcatc agctcctggc cacaaagtca ccttgagctc cactgcccta
2521 ttcacaggtc accccacttc tctgtctctc atgagccctt ttcccttatc cactgtgtct
2581 tcaagatcca cagtgaaggc tgggacatca ggtaggtcgt aaccctgggg tcttagtctc
2641 ttgcacttgc ccagttggct caattcagtg cagaagagtg cttgagagag gacattcaat
2701 tagcactgcc aggctggagg tcaggtccag aaggttatga aatcagttgg ggtttggtgg
2761 gtgatagcct caccatcctc tttaccccag gtggcagctg cttctcctca ggcagtggtt
2821 ttggtgtgac tagaagacag ttttctagct gtctcttcca ggttctaaag tctatgactc
2881 tgtttaactc tgctaccccc tgactgttag actcactcct gtcttctctc cctgcctgcc
2941 taggaacaatcaccaccttg cagaagacag acagcaagacaagcacagca gcatctctac
3001 cctcaaccac ctcccaggct cttacaacttccactgctca cacttccaca ggcactcgct
3061 caacagctgc cccagtccct cccaagccga agaaaggtga gtgaatccat agagctgggt
3121 cctcttcacc ctgggtcaaa gaggatggag ctatatgtgg aggttatagt tgtggcagct
3181 ggactggatc taccaatgag gatctttgtg gggaggtagg gggaccagtt ctgtggatcg
3241 ctccactgcc ctgggtgtgg caagtccagc tgctctctgt gtccccttga taggggtggt
3301 gtaactgagt gctttgtctc agcctgcttg cttacagact ctcgagtcag aaaggcctta
3361 cagtgaacaa tgtgacatca tcagtgttgg gtgctgaggg ctgactttgc acatggtgct
3421 aggctaagag ctggagataa aaaaatgatt gagtcactgt ttagtgagac agccttgctg
3481 cctcagtagg tgcgattaga ggcataaacc cagggtatta ggtacacaca atggaaggcc
3541 ccaagggaat attggggtgt gggttcaagg tgaaaaggag tctccctgga aagggtgaat
3601 aggatctgcg gggaggggaa gtgccccaag caggcaggag ggcctgaata gctagaataa
3661 aggctcggac aggaagagca gtcctagtat tgcatgggac agagatcccc atagcatttg
3721 gggctgccag cagtaaggtg aaaggtgttg agaagcaagg ttgttatgtg aaggaatctt
3781 gtagatgaca ggaaccaggg agaatgtggc tagatttgca tttaggaaca gtgtcctcat
3841 ggctgggatg gaggatgaac ttgcaggagg ccagggcagc agggtagata tcactgaggt
3901 tttccaagca agaccaggtg aaggctgcca gtatggaaag gaggaggccc ctttgagaaa
3961 tgtttgggac aaagtttctc aaaccggagc ccaagggcct cggtagggga tcttatgaca
4021 aattctagca gggggcagtg tgtgtgtgtg gcgggggcgg tcttcaagtt tacctagaga
4081 cataacatct taggaaacaa gtctcattct gtgaaacgat cttgcaatta ctttactgca
4141 ttagaggcat tttgtctttt aacatttcac agaattgtga gtaccgtaat aggtgcagaa
4201 taatgtgggg gtgttctgat agctgttttt cttcaaactg gggaaggcgg agagtaccat
4261 gtaagggagt gaaagaggag atgggttaaa tgcaagtgga gagggggaga gagaagtcct
4321 tgacggctct tgcttaagcg aacaggtcca agggtggagg agcagcttgg ggacactctt
4381 gcgggacccc tgccagccct aatgacatca gggcccatga gaacttcctg cttctctttg
4441 tactggccca ggcgttcccc tcttccccta cgggccgcag gctggagact ggcagtttgt
4501 gagaaggacg gtggacttca cctcgccact cttcaagccc cagattggct tccccctcgg
4561 ttcctctctc cgggattgcc tctacgtgag tccttggctg cagcccgaag gcggggggcg
4621 gggggcatga cctgcattcc tcaggccttt cttctccctc cagggctcag agatggccct
4681 gtgatcatgg ccataactgt gactgtgatt gacagggaga acggaagcaa cgcttgtcag
4741 gcacctgctg tgccccagtt gctgcctcag tttgcaccat ctttaacgat gggcaccagc
4801 tgaggaagac agggagctag gcgaggcccc ttacccacgt ggacagaagt gtggccacct
4861 cagtagagag gtcctggggc ccagtaagct ctccactgtt ttcatgggtc cttttttaca
4921 agtaggaagc tctgctgggg aaacctgtat gatgctgttc acacggtctc tcattgtttc
4981 ctggttccct agtccaagta gcccatgtct ctggggggag agcagaacag cagtcgtggc
5041 cctcgagagg gctggtcagg acacactaca aaagggtgac tcagcaatgg ggtcagggta
5101 tcctttagcc ctaattgatg tggcacaagc ttcagggtgg gctgagaggc tttactctgc
5161 tctgggatgg gctggtctct agacgggtgc cactgagaca ggacaaaact accaccttag
5221 gaaagactca ggacataaaa aaaatgtgct gagggtgaag agtatttgtg tgagaactat
5281 atacagaggc tgtcccagca tgcagggaga tggatgagac cttccagagg ttgttggcat
5341 ctctctggtt ctacccagca gggcccttca gggtcttgca tcagtccagg agctggcccc
5401 aagccgaggc tccttcccca cctcccgccc tgaatgctaa ccaggagaac ctttgtgttc
5461 agttcacaga taatggccaa atcattttcc cagagtcaga gtaccagatt ttctcctact
5521 ccaaccctcc cctcagaggc tttacaggcc aggaccctgt ggccctggtg gctccattct
5581 gggatgatgc tgatttctcc aggcaaggaa ccacgtttta ccaggtgggc ctttcaaagc
5641 tcagcattca gggtcctcca gcagccagca aggacagaca gagggctgtg tggagggcct
5701 tgcaagtgtt actgctgtca gagcctaggctcagtgtggg cagggactaa aacttaaatg
5761 tgaggatgag gaggaagcag atgaggacaa aatcatggga agactgaagg tgccagggat
5821 ggggattggg gatccatcgt ttgattttag gatgaggttg tagcagggtc cctcctaata
5881 aggctgagtt aagcaaggcc cctgcattgg gctgatgatt ccattcagtg ggagctattg
5941 tgtaatgaaa gctaccacag gcaaaggtac gtcgtgcccc tgccaaggac caggcactat
6001 agaaggtcca aagatggaga aacatgtgcc tgtcctcaag gagcctccag tgggacacag
6061 ggcgatacag atgggcacag gtggctgcac aatccaagcc agtctgtgct aagcgaggca
6121 caagttcaag tccagaagtt actggcctcg actctccact ccacataccc caggtttccc
6181 tgcagcccga acctgatttg atcttaaggt cagaggtgct aactgtgaaa atgtgggcct
6241 gggacggggg aggatggaag atatgggcag actcaaatga gcaggatcac catgcagaaa
6301 gtggggggtc gcccaggagt gttgagctgg gatccttgtt tcttgtctga atctgccttc
6361 ttttctagga gtatgagacg ctctatgatg aatacaacca gctagtgtgg gaggtggagt
6421 attggattaa aatgttaaca aacacctgga actacaaagc caggtggaca ctaaaggtca
6481 cgtgggtcca cgcccctgcc tatcctgccc agaagaccct cagggtgagt ggaccagtgg
6541 gcagctccct gggaaccatc tggaagtcag acatcctaga atcctaggca ggggccactc
6601 ttctaaaatc ctttcggact cttacagaac ctcacactag cccagagcac tgggtattag
6661 gaggttgaaa catagaacgt ggaatgtcca taagtcaaaa atggctggca attggcaatt
6721 ctatatggct caacctaata tttttatcca catattagaa acgaggagtg ccctggtggc
6781 tcagcaggtt aaggatctgg cattgtcact gctgtggctc tggttgctgc cgtggcacag
6841 gttcgatccc tgatctgaca attttctgaa tgccaagggt acaacctaaa taaataaata
6901 gaaatagaaa gaaaatagcc agaaaatata actgtgaatc aggtagtcaa aatacatatt
6961 tttttaaaag aacgaaatga ggaagatgat ggttagaaaa gttaagtgac ttgcttcaag
7021 gtcacccagc tagcaaatga caaagccagg cctgaaacag aggtttttct gactctgcac
7081 tccagctctt gaccaaaacg catccactgg aagcaggcac accacatgtc cccccctatc
7141 tggggcctca ccaccctgat agtcacaaag ttcttccttg tgtctggtct aaagtcactc
7201 atttcttcat tggctcaaca agctcattgc ttacctactt tgcacaagca cagtgctggc
7261 cccggccttc agagaacctc cactctcacc tgtgtggtcc tccacgccat ccaggtggag
7321 gggacatttg gacaatatcc ttaaatatcc ctgaggaagt tccttaggcc tcccccagct
7381 ctccaacgtc cgctgatcac tcaggttgct gaagctttct catacaaacc ctcttctcac
7441 tgaattctgg cctagagtcc tttggctccc ttcctgggga agaagtccca aagaaggcaa
7501 atccaccatg gcaggtgctt ggacagtcag agtgaaatct ctgcataatt ccaaatttgc
7561 ccatatttta agcccacacc ttagctctgg gtaggccagg aaacagtgat atgggtgtca
7621 tgcccaggct gtgtgacagg taatgggaca taccagatct ctcacgggca tccctatgct
7681 tgtctccaac caatccccag accaacacct accaggccat cctttccacg gatgggagca
7741 ggtcctacgc cttgtttctc taccaaagcg gtggcatgca gtgggatgtg acccaacacc
7801 taggcaaccc cgtcgtcatg ggcttctcca ggtaggacag agtggggctg tctgcattga
7861 gtagacagca ggaggcggtg caaagcaggg cttgcagctg cacacgaact ccattcccca
7921 aggatacgcc aaaatcctcc ctgcccatgc ttctgattca cgctgactcc agttcccact
7981 tcttgctgtt tcaaattaaa agacccctat tcactctgcc gttctctttc ctatcaaggt
8041 ctccgtgcat gtgataataa tccccagagc cttgacttct aggacttcca ctgacagtag
8101 ctgacctcac ttgacttttc agagcttaac aataacagcc aacatttatt ataccagcac
8161 tcagtatgca ttattttgta tattacaaca accccatgaa ggagatctat tttattaccc
8221 attctacaga tacagaaact gaggtacaaa tagataagtc attgcccaaa gtcacagagc
8281 tgggatgtgg caaagtggga agctgaaccc aaatataagg ttgaaatctg ccttcctgac
8341 tttcgattgg ctggaaatta acctctcacc caaggcacca gcagcgtctg tcagtggcaa
8401 agatggagaa cagagagatg gatggggacg tgggttccca agggaggagg tggctcagtc
8461 tctgacacaa cagttctcaa ccttgccatg tgcgccactg gctgtgaagg tcgcatgtaa
8521 tttgttactg ctaattgcta caaatgtttt tctcttagag tgtattcagg attgcgcctg
8581 taattacatt gctttcactt gcttagtgat gagccctgaa ggaaaaagaa gtagaattat
8641 ccgaccagct aggaatttca caccattctg agtgcaccca cggaaggtgt cacacccaag
8701 tgcgtcatct gtcctgtgtc agagaggaga aaagatcagg agtggctgct ctgaggcgtg
8761 caccctgagg ctgtgggctt tgctccttcc ttccaggggt gggggtgggg cacacccatt
8821 gcctggtcct cactcggtgc tgagggattt accagtgtta ctgctttgaa tcttcacaat
8881 actgtcaccc ctttgacaca gaggtgaaaa ctaagaatca aagaggttaa attaacttgc
8941 ccatgatcac caagctacta agtagtggaa ctgggggttg aatccatgac cgagagactg
9001 gcttttaacc ctcacagcca gctgtgccct gcagcacccc ttctccaatt cctagttgaa
9061 gggaaactaa caggaagggg gcctgacctg ttgttctcac acccctgtct catgacccgt
9121 ttcctacagt tcaccatcag atgtggaatc aattagagtc tcctattcct gccccaagcc
9181 tcccttcccc actcctgctt ggcaccgggt ctatgattct ggctggctga cgaaacgact
9241 gaacagctga taactaatga ctggctggac tgagtgggtg accagctgac tgaccagctg
9301 tatgaatgag taacttgctg actgagagga aaaactgtgt gacagtggct gaagattctt
9361 ccttttgcct acagtggaga tgggtatttc aaaaacagcc aactgacatt gcagccagtg
9421 tgggagaagt atcgtccaga ccagttcctg gatcccacat caggtaaaag caccacctta
9481 tcacacttga gggacccaag tccccttctg tcctcccctc catgtgccat gacagtccca
9541 gagggcagct cccttgcatc tccaccatcc caaagggtgt ttccagcaca actgacttcc
9601 catggtgcaa tccagatgtg gcactgccca gggagtggtt acctcccgtg ccctcacaca
9661 gcacaaagtc atttgaggct gcttctcctc cctgggcgga gccaaacaca ggagagctat
9721 gagtgggggc cctgatcctg gaagtgagga caggggcatg gcaggaggtg aggaatgaat
9781 gggcagctag gacaattctc caagtgtcct gtcccagaac agtaatgaga cttcattctt
9841 caatggaatc taccaagagg agctccaggg gatctgacat ctattcagtc ttcgtttatt
9901 gagtacctgc tgggtgtagg acactgtgct aagagccaga agtatgagaa atgaaaaaga
9961 ctatgtttgc tgtcaaagag ctccccctct ggtaagaagg ccagtactct aggccccacg
10021 ggcctcctcc atgatttgat tactgtccgg tgacccctaa agaaaagaaa tttcagtttc
10081 caaatgtgac agaacaaagg atattctggc attaaaaatc tgccatgtaa agtatgaagc
10141 attaatccag ttctgattgc tctgtggcaa caacttctga taaacagaag ttaagtcaaa
10201 tcatgggtgc catcttgtgg agaaagctgt cctattagac tagggccctt ctatccagtt
10261 tgtgcttgta gggggcgcaa taggatagtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
10321 cgtgtgcatg agcgtgcatg taaggcccct ctatccagat gcatctgctt gtgacgtggg
10381 gatgcaatag gagtgtgtgt gtgtgtgtgt gtgtgtgttg ggggagagta tctggtgtgt
10441 ttcccctgat accaggccac agtggagaca atgcacagat gatcataaac aatgattact
10501 cttttccaaa cctcatcaaa actgtagtta gcttgatact ttcttttaca cggggtcagg
10561 gtcaaaatca aatacattat atcctattgt attttactgg actctggagc ccacactcag
10621 ggtttggctc tgagttggat tacaggtgag gccctcctgc ccagcacttg tctcaaggtt
10681 tcgctgtaag atccaaactc ttgaggtttt ggaggtcaca tcaaagggcc tttggggagg
10741 cactactgcc agtctgactg ggtccattcc tagaagagct gtccagggca gggggagtga
10801 catacagagg gtgcatttat gagttttgtt tgttcccaaa actaaagcag agcctgtggc
10861 ccaggccctg agatcacccc cttttcctct gctccccagg cctccggggg ctgcagatct
10921 ataagctaca caaggaagac aagcccaact atcgtctctg gtgcctgcgg tggttgaaga
10981 gacagtctca ctggtccggc tggggctgga accaggtctc ctgcccctgc tcctggcagc
11041 agggactatg ggacttaaga tttcagccca tcaacatagg tgacacctcc ttcatagtat
11101 cctgtggtgc cccactgtcc ctcacaagcc tgcccaccct ccctttggcc ttgcctgacc
11161 cgtctcttcc agggcaccct cagttcctcc ccagaagcac cccctaccgt tgatgcacac
11221 accaaaggca tcctgggcct gagtgtgtct ggcttgttgc aggctggtgg ggcatcgtca
11281 acaggcagct gtgcagcttc tcctcctggc gtgggagtgt gtgctgcagc tacgggccct
11341 ggggagagct tcttgaaggc tggagattgc agagtccttg gcagtttggt atgtgtgtcc
11401 acagagaact tgagcccagc ttttctggag ctttctgttcttccctgtca gagaccagcc
11461 agcagtgaga cccacagaag gaaggctctt gtcccatttc ctcccagcta ccttccccct
11521 ctggccttca atttcctgat ctggtaacat gaggaaattt ccaacatccc ttccagctga
11581 caccctaggg atttatgatg gagtccctct tgtcccatct ccacttcgcc aaaagcccca
11641 gcccagagtc gggcatcttc tagggaagag aggggcctgg tctggccctg tccatcggaa
11701 cccagattag cagctgtgca ggggaagact tggagtcgcc tcctgcagag accctccaaa
11761 gcagctagag aggacagcgt tggggggcca gggtcatgcg tgtccttctc acctcctcct
11821 tgtaatcaca gcctccttag tccccacacc accagccatg ccctcctgct cccacagcac
11881 tttgagactc agcaccccac ccttcaagaa aggaagggcc ctgggagttc ccactgtggt
11941 tcagtggtaa tgaacctgac tagtatccat gaggacacag gttcaatccc tggcctcact
12001 cagtgggtta acaatctggt gttgctgtaa gctgtggtgt aggtcgcaga cacgacttgg
12061 atctggtgtt gctgtggctg tggtgtagac ctgcagctgc agctccgatt tgacccccag
12121 cctgggaacc tctatatgcc gcaggtgcgg ctctaaaaag acaacaacaa cgacaacaaa
12181 gaaagaaaga aaggaagggt cctgtgcctg gcccactagg ggctttgcta cacatggctc
12241 caaataccca caagggagac agtcactggt caatggcttc aggtcattga gcagggtggg
12301 ttctgaactg aggtgtgccc gattcctaaa ccataccctt gccaccgttt catcctgcca
12361 gaactgccca gtgctacttc ctgcaggttc tccatctccc ccaacacgct gctcttcttc
12421 acagaccaag aactggagcc ccagaactgg tgctgccgct tcaatgacaa gccctccttc
12481 tgtgccctgt atgagctaat gcggccccga atcggctgca gtgggtacca gcccccgagg
12541 cctggtgagc tgctggagcc tggctccagg acagggcaga gacagagcat caggggaagg
12601 gtagcctctc aggacctgac caaggcagca caagcttcct ggggcagggc tgtggccaca
12661 cgggaggatg gagatgggtc cagtgtgtgg agatggcaaa cgagaaattt tatatgtggc
12721 cttgggggag gggaggtgtt gggggtggga aaatgaagat gcggtggggcgcccaagggc
12781 aggaggaatt ctggggaggg gtctgtgctc catgagcacc tccccaggct ggagtctcct
12841 gagcaaacat cctgtctggg gccagtaagg aatcccaggc ccttccctgg tccactctgc
12901 caatgtggta tctcctcttt tccagcctgg atgtttgggg acccccacat caccaccttg
12961 gatggtgcca attacacctt caatgggctg ggggacttcc tgctggtccg ggcccaggac
13021 agaaactcct ccttccagct gcagggtcga actgcccaga ccctctcagc ccgggccacc
13081 aacttcattg cctttgctgc atattacagc tccagcagcc tggaccccat cacggtgagc
13141 cttcagctgg caattgagtc ccaggaggag aaagagaagg aagaagggca gggggcagag
13201 gtgagggagg atgcatcctg ctggcacagc agttccagag tcaaccctgg aaacatagga
13261 ttagggtctt ggctccacca gctgtgagac tgaagaaagg acctactttt cagtgcctcc
13321 cagtttttct tagggtaaaa tggggacgat actactacct acacacgggg ctgttgtgag
13381 atctaaaagc taatctaaga acccgtgcca agggcctctt acacagtggc tagcatcacc
13441 tcatcgtcac actcccagtc accttcgctt tcatccaagc cccaggctct aagctctcgg
13501 aagccttctt ccttggcctc tggacccacc tgcaggtcgg gcttaggcct ctgccaattc
13561 ccttccaggt tcaatggctc cttgagccca atgacacaat ccaggttctg ctcaataacc
13621 agactgtaac atttgagact aaccatgcag gtgccgaagg taggctgggg atggccgtca
13681 gctgccctct cctgcttccc tgggtcgggt cactggggag cagggtgctg gggtggagag
13741 gcagcagcat cgggcagagc agagtggtcg ccacccctgc tgtccttcca gcagctgtgc
13801 ctgagcagct tcttcactgg agccgagcat tgagcttttc ctccgcactt ctctccttcc
13861 tccagggctg tgacggcctc tctgggcaat gccagggaga actggttggg ggggcgggtg
13921 ccatgagggt gtcctgtgag cttggtttgc ggctactggt gtaggacctg gcagggcatc
13981 tcccagctgc tctcagcctc ctctccacat cttgaacctt cactactgcc aagtcttggg
14041 cctctaggac tccaacactg gctgtttcat catctgtctc cccaccttct aggccaggag
14101 atgttcaaca cctctggagt cataatgatc cgcaatggct ccacggtgtc agccagcttt
14161 gatggggcgg tgaccatctc agtgatcgct ctcgcccaaa tcctccatgc ctcctgcggc
14221 ctcccaaagg agtatcagaa ccacacagag ggcctcatgg gtaaggagcg ggcaggcccc
14281 tgcctctgca gcaccagctc agacccccgg cctacaggca gctccccctc ccgtgggtct
14341 gcggatactc cgggtggagt ttcctttttc ggcctcctgg gagcagagcc ctgagggtac
14401 agagcagcag ggagggaggg gggacccctt cagctacctt tgaggctcac ttcccatcct
14461 tctgaagctc tggttccagg gcagccaaga aggaatcaat agtgggctct gggagttccc
14521 attgtggctc agcagtttaa ggacccagtg ttgtctttgt gagaatgcgg gtttgaaccc
14581 tggcctcact cagtgagtta aggactggca ttgccacaag cagctgcagc gcctattcta
14641 ttcaacccct agcctgggaa cttccatata ctgcaggtgt ggccataaaa agagaaaaaa
14701 aaaagaatgg gctctgactg ccttgggtgg cggggggggg gggaggaaag gggactgatg
14761 gagagtttga gttggtaggt gtaaactatc acattcagaa tggataagcc ggggggtcct
14821 tttttccccc cccgggaact atatcccatt ttttggggta gaccatgatg gaagattttt
14881 ttagaaaaaa ctttttttaa gtaagggggg ggggggccct tttggggggc cgccgaaatt
14941 gccccccccc cgtaaatcaa ctatatgcta attttaaaaa aaaaaaaaaa aagaatgggc
15001 tctgggaatc atgggaccag cagccacccc accccccact ctggctttat acctcccacc
15061 ctccctccat cctccgcatt ccctcccttctttctccttt cctccttcat ttccttcttt
15121 cctgaagcgg aggggtgggt tctgaggtga cagcccgttc ccctcctcct ggaggactga
15181 atggagaaga gatacaatgg acggaggccc cctactcctg cacgcatctc catgtgacca
15241 gccggatgct cccagggaag ggcaaagcca cagtcccagg atggctgact cctgctctgt
15301 ccctcagggt tctggaacgg caacccagat gatgacttca ggatgcccaa tggctctact
15361 ctttccccac ggagctccga ggagaccctt tttcagtatg gaatgacctg tgagtctggt
15421 cctcagagtc ctcgggtaga tgggcagggc tgctgcatgg tctccctcag gcctgtggga
15481 accagacctc tgtcctttgt gagagagggg tagaaaggaa gagatctaaa gatgctggtg
15541 ctacccccag atttgtcccc tctgtcccca acccacctgc gcctctctac ctgggagagg
15601 ggacaaggga ggtgggtggc cctcagtcac tagagtgagg atttccagac tcagaagctg
15661 gaagtcactc tgtctctcca catcccctgc cccaacccca aaccatataa gggtaagggg
15721 tggacacaac agaaatacag ccagaggccc accctggtaa caccaccatc atgccttgtg
15781 tgttttaggg gagatcaacg gaaccagcct ccttggcaag aggaatgacc atgtgtctta
15841 taacttcacc cctgtcttcc tttcacaact tcggggaaac aagtccttga ataaaagttt
15901 gacttccagg tgtaatggag atgaacaatg catctatgat gccctggcca cagaaaatgc
15961 aaaccttgga gagcacacta tgcggctctt tcgaagctac cagcaaatga atgctacctt
16021 gagtgagtgg cgtgaggctc ggggaggtgt gtgcagagtt agggggcaga tgaggagccc
16081 ctcttccaag acacccctta agctacacat ttttttccta aagttattca acattttaat
16141 cccctcctaa acataacata aatttttggg gggccacacc catggcatgt ggaagttccc
16201 aagcggggaa tcgaacctgc cccacagcag tgacccaagc tgctgcagtg acaatgcagg
16261 atccttaacc tgttgcacca ccaagagaac tcctaagcta cacatttttt aatttagccg
16321 attattttgc tcatgttgtc cctctgctca aatcctcccc tgactcccca gctcactcta
16381 tccttatgac aaccttcaaa tcaaagccct gtgcactcca gccccactgc ctctccacac
16441 tcttctccta gaacttctag aacctcgctc cctccactgc atccccactg cttcttcatg
16501 ttcttcaaac acgccaggct cgcccccgac tctgagactc tgcacatgtt ctgttctctt
16561 tgcctagaat accttccccc agacacccac aggtcgctcc tcacctcctt caagtctcta
16621 ttaaatgcca ccctctcagc agtgcctttc ttggctaact tgtctaaagt ttcaccttca
16681 ctcacacttc ctatttgcct tccctgattt attttttctc tttagcatct ataactatct
16741 aatgtatttt ctaattgtag caggtctttt tattatttgt tcattgctat gcccttacga
16801 cctagaacaa tgccagccat agagtaggtg ctcaataaat atttgctaag tgactaactg
16861 aatgaatgaa tgaacaacag aatggaaata ccagggctcc aaatccagag accagtggaa
16921 aacttttact ctctcattct acttcctgac aatgtaatat ccacctagat ataaccctca
16981 gtgtttccag ctctcctcca aaggtaaaat tccacaatgg cctgaacagc tctgtcctct
17041 gatcacttca taccatgccc tgcataactt tctcaccagg aaagaacctg aaatgaatct
17101 tgtttgggga gaaaataaag agagagtaaa gatttggctg ggtgggcaat acaacaatga
17161 cctgctctgc atagatgcaa gagcaggtgc atagattatt tcctgcttct ggaggtgggt
17221 cccagccctg ggaatgcctc acacaagcga gaagactaga gagagagaaa ggaggcccag
17281 aaggcaggaa atatcctgcc ccagaccctg accccttctt tgtgtctcag atcagtaccc
17341 gccctccatc aagggtcctg atgtggtgaa aacctatatg ggacaggcca gccttgttaa
17401 ttacaccagc agctctgaga acgtcacatt cactctcaga cacaactgca ctgacttcaa
17461 gctctctggt aagaatgctc ggctggggca gttagtggtt gaggtctgag tcaagctttg
17521 ggtaattttg cgttattata ctctgtgcaa cttcatgtca cctttcattg gaatatctag
17581 gaccaacatt acccaggaat gatccatata aatcaaatag attcccccca gagattaata
17641 gtgatctggc cagctagctc gactgactcc aggctgattc aaagatcatt tcttctcatt
17701 cttgtaatat tgcaaaatca tctttatgca atcaccaagt tattagaata aatttttttt
17761 tgtctttttg tctttttagg actgcactgg tggcatatgg aggtttccag gctaggggtc
17821 aaatcagagc tgtagctgcc aacctacacc acagccacag caacatggga tccttaaccc
17881 actgagcaag gccagggatc taacccacaa cctcatggtt cctagttgga ttcgtttcca
17941 ctgcgccaca acgggaactc ctagaataaa tatagcaatt aaaaagtact aatttaaaat
18001 atttgcacat ctattaaaag aaggatatca tatggaacaa gatttaaaga aatacaaata
18061 ccttcatttg gtaactttct cacaattttg atgtttgtca accatgttta cctaactttg
18121 cccctccttg cctgcttttc tgtactgggc ccaagcagtt tcactttccc tctcagtaat
18181 ggtgtggaga ccccatttca gaacgggaac catgatctgg gtcaagagtc aaagcacaca
18241 ggttacagct tgccttttag ggccagccct aattaagctg gtgactgctc tttgctaagc
18301 cagagccctt agggaccttg ctctgagcac ctctctgcct ctctttctct gtgcataaga
18361 agagacagca ttgtctgcac cctgtgctcc atgtggcatg ctccctagtg cctaatcaat
18421 ggcatcagga gactcactcc tccactgggc tcctatcctg aagagggagg tcctggaagt
18481 gactctcatc tgctcccacc catcagctct tttcaatcca ctgttttaag gagacttccc
18541 tattttcttg aaaaacataa gacaggttga atcagaagag tccttattga tacacagcag
18601 tatttccctg tagcctgggg tctggtcaaa ggtccaggaa atgagagcaa gtgggtgagg
18661 gatgaggctg ctgtgaagct gaggtcagtc tggcctgaag tgtagcctga gtaagtgttg
18721 ggaagatcct ggccttcacc ctccatggct ccccgccccc catatcccat cctagtgcag
18781 gtccaggcag ggttaccctt tgggggcttt ggggaggtgg ggggcaacca ccttcatctc
18841 atggagttct accctagaat cacttagttg ggctggtgcc acctccaggt tggagtgtga
18901 tccaacagaa actagaaacc agccttgctc aaggaaatgc ctagtaagat tggaggggtg
18961 gtctgacagg ggtgggggac gtccagcatc agcacttggg gtattagatg ccaggatgtg
19021 tcaggcagca gaggagatga ggctcccagc tctcaaaatg atctgtcttc cctgggggca
19081 gagaatggga cgttgctatg gacaccacag tcactgaaac catgtcctct ggagattctg
19141 gcaagaagtg ccaaggatga cttgtcatct gtactcaagc caagaatggt ggtctgcact
19201 tgccaggcag agagccagtg tttatataac cagaacgatc gggtgggcaa ttcctccctg
19261 gaggtgagtg tgggagaggg tgggaggtcg gtctctgtgt tgagggaagg gaagtgggaa
19321 ttgaagggat gttgtcattc agcccctctg tcctaatgtg tgtgttggga ggtgggtgga
19381 gctgtggtag gtgagtaggg cagataccaa ttttgggcca caaacataag aaataggact
19441 ttctttagtg cttataggtc aactgaattt ttacctccca gctcccatcc agccaagc ta
19501 gcaggaaact tccacctcag cctttctttg agttgctcag tggtttctag ctccatggtt
19561 ctggaacgag acaatgaagt gtggggtggg aggaggtcca tgtggcccct ggggggcctg
19621 agtctccatg gatcacagct tcttcatccc atcagatgta gggagatgct ctgtggcttc
19681 tgggccttac ttggatacag gaaacctcct ttaaagtggc ttagggtccc atgtgcctag
19741 aagcagtgca cagccatttc tttggggcct catcgtcatc ttgggaagag aggtacaggc
19801 cccctctgat tttaacacct gtaaaatcct atttttcctt tttgctctct tccaactaaa
19861 cgaagcatct ctttggcagt ttagagaaaa tccttctcca ttcaagtttc tagccaagat
19921 acttggccta agtccatgaa tctggcccta tctgcctttt tttttttaat ggccatgccc
19981 acagtgtatg ggtgtgacca ttaaaaaaaa aaaaaaaaaa acacttctgg gccagggatt
20041 gaatctgagc cacagctccg gcaataccaa atcctttaac tcactgtgcc tgtctggaga
20101 ttgaacccac acctctgcag tgacccaagc tgctgcagcc agattcttat ctcactgcac
20161 tacagcagga actcccccta cctgcttctt aaaaaaaaaa aaaaatgttg ttgttgtgaa
20221 aggggagctg gaaaagacag gttttgactg gcaatactgc accccattct tcaacccctt
20281 acatacacac acaccctcct tatgaaacat gaacacagaa ttctgacttg aaagggacag
20341 aggtggggga gtccaggagg cagaaaaaga aaaaaaaaaa tcagaatgga gaaaggaggg
20401 attaaaatca atacattatt ttggatttcc caagatatcc cttcttggca gtaggaagtg
20461 ttgaagggct ccatttcagg gagctttcca gagccgtctc agtagaggag ttagagagac
20521 ccggagcttg gtaaagtttc cgaagggttt ccctccctgt gcccttgctc tggtggccat
20581 gtgctctttc agaaggttct atcttggagt tcccattgtg gctcagcaga aacgagtctg
20641 actagcattc aagaggacac agcttcgatc cctggccttg ttcaatgggg ttaaagatcc
20701 agcattgccc tgagctgtgg tgtaggtcgc agctgtagct ctgattggat ccctagcctg
20761 ggaacctcca tatgtcacag atgcagccct aaaaagacga aaaaaaaaaa agaaaagaaa
20821 agaaacaaac aaacaaacac aagaagaaga aggcggctct gtcttcttcg aagtcagcaa
20881 ggccagggag agcaggacac tacagcttgt gtgggataca gctctccaga aagaatagaa
20941 tatttattta tttcgcccct gccttattct agaaaggaatt cagtccagg agagaaggtg
21001 tgagagggaa gaggcttgag gtcccagggc aggggccagg cagagggctc cggggaaatg
21061 atggcccagg gaggccaccc agcttggagg atgatcggcc tttgggcagg gatgtggagg
21121 cccagggtct gatccaaaag gtgctcagtg atactgacag aagacaagga cctgagtcca
21181 tgggttctca gaccagagga gttggtacag ctacacccag aagcccttga tggttcggct
21241 tcctggcggc ccagtgatca gcaacccatt ccagatggcc ggctgcaagt gtgacgggaa
21301 cacctttggc cgctactgca accactccaa ggacccctgt gatgagccgt gcttcccgaa
21361 tgtgaagtgc atttccggga agggctgcga ggcctgcccg acaaccctga ctggggatgg
21421 acgtcactgt gcacgtgagc tggggacagg gccttggcag gaggaggttc tggggatggg
21481 gcctaaacag ccaggaaaaa ccatttctcc cccctttcca gcaagtcttt agaatgttcg
21541 ataaaggggg ctatggtaag gaggtaaaca ggggagggga gggtgtgagg ccaaaggagg
21601 agagatggtc caggcatggg ggaggggcca gagtacagca gtcccaaggc aacgatgggg
21661 gaaaccaggg caaaagactg gagacaggga gtttccgtcg tggcgcagtg gttaacgaat
21721 ccgactagga accatgaggt tgcgggttcg atccctgtcc ttgctcagtg ggttaaggat
21781 ccagcgttgc cgtgagctgt ggtgtaggtt gcagacgcgg ctcggatcct gcgttgctgt
21841 ggctctggcg taggccagtg gctgcagctc cgattcgacc cctagcctgg gaacctccat
21901 atgccgcgag agcagcccaa agaaatagca aaaagaccaa aaaaaaaaaa aaaaaaaaaa
21961 aaagactgga gacaggaggg gacttccagc agcacccagg ggaccagagg taagctggct
22021 cctggaggtt tccagagatg caaggcccag ggcctctgca ttctccagtc aggacactgg
22081 cggcagggtg gggtgggggg gtctgccatg ggcatctagc agcaggaatg ccttgtcttc
22141 cctttgagtc ctccatccac catctgttcc ttgacctctg agccactgga gtcatgggtg
22201 gcatcatggt gacatcttca ctgagccccc taaaacactc aaatgaaatc tagaaaccac
22261 tgcatcttct gagcacagcc cagattggag aagcacagcc ttataaaggc caaaaagggg
22321 atggaagtga ggaaagcagg ctggtgggcc tttctcctcc cagccttttg ggtcctttcc
22381 tgatggggag ggtctaacaa cactccattc cctcagcaga agtgggagaa cccctacctg
22441 gcagaggaaa aagggttgtc tggcaaagaa atgtctggaa aagggagaaa gatggtttgg
22501 ggttcagagt ggcccacccg cagaacacag tttaagtgag ctttgcaggg aaaaagccaa
22561 ccctgtcctg ggagcagctt tctctggctg gcggctgcca gtgtggtgta gcagactgtc
22621 tgttccctct gcagctctca acagcttttt caactgttgg aacaagacct gtcctgagaa
22681 ttactgctac aaccagggtc gctgctacat ctcccagtct ctgacctgcg agcccgcctg
22741 cacctgcccc ccagccttca tggaagacac ccgctgcttc ctggctggaa aaaatttcac
22801 tccaaccatc cttccaggta cagccagatg ccctgccatc cccactccac acgtctctta
22861 acttggggga agatggggga ctctgggggc caggctcagt ccctcaaact cacaggacag
22921 aactccaaat ctctctcgtg tgctcgtaag ttgtaggctg tgggagtgag tggggggcag
22981 ctgtcacacc acaaaccaca ggtctgagca gtctgctgag aaaagactga aggggacctc
23041 acagctcagc gagacaagag tcaggccagc cttgatcaca gggtgaggtc agcagacctg
23101 gccatttctg attcttctgg tgaccttcta tgactgtaga agagggcagt cttcttgggc
23161 tgtcctcctt ggctaagtct gggtaaacct gtgaggtgac tatgaatctg cttcttccct
23221 ctccagagct tcccccaagg ctcatccagc tcttgctcag tgaaaaagaa aatgcctccc
23281 aagcagatgt caatgccacg gtcagtgctg caggctagctctgggtgggg aggcggttct
23341 tggcagattc ttatccatgg cttgtacatt ctgcatctca tcatccatct agactcaact
23401 gccagaggtg gctagagcct cctgccccag gggggtgggg gtgagggagg cagggaggcg
23461 caggacagag acctccaaag ggctgtgaat gaccatctca caatgaacag aatctggggg
23521 cagggagagg ggggcaagtc aacacagcct cgcctctcct tctgagcatc cctcgagacc
23581 tgctcttggg cactatatgg aagaaacaca agagggcatt tcacctccag gaagccttcc
23641 ctgaattccc caagagaact gggggctcct tcttcttggc tcccacaccc ctgttgattc
23701 tgcagtcaca ggcaaagtag cacaattgtt gagttactga ggctctagaa tcctacctgg
23761 gttcacatcc tggcagtacc actcactaga gcaggtatga tcattagaca aattatttga
23821 tctcccatat ataaaaacag aagtaataat agtactcatg tcaatgggtt gtgtcaaagt
23881 tgaaatgata atatatgtaa agcacttagc acagtgcctg aaacacaata cattcaaaaa
23941 ttaccaaaat aactattgtt tctgctgcta ctgctactac tactacaaca gcagcattct
24001 gggatctcta tggtaaaggc tgttgacatg gatctttgag tatgggcagc attctgggat
24061 ctctatggta aaggctgttg acatggatct ttgagtatgg gctgtttcta actcacactg
24121 acacacacac cccccccacc ccaacgcaca catctgacca tcccccaggt ctggctaaat
24181 ataattgatg ttcattcaga tatggggaga gggtgagtgg gtgagtcagg gatcagaggc
24241 aaggtagggg ctgttcattt catattctcg ctttgtgagt ggaactgggg tgaggttatg
24301 taaagtccta cactcaccaa caaacttcta ttaagtaggg acccatgcca gtcaccagtc
24361 gaggcttgtg gggcggggat gcagaaatag ataagaaaca ttctgagatt tcccatcatg
24421 gctcagtggt taacaagtcc gactaagaac catggggttg tgggtttgat ccctggcctt
24481 gctcagtggg ttaaggatcc ggcattgccg tgagctgtgg tgtaggttgc agacatggct
24541 cgggtcccac gttgctgtgg ctctagcgta ggccagtggc tacagctccc attcgacccc
24601 tagcctggga acctccatat gctgcgggag cagcccaaga aatggcaaaa agacgaacaa
24661 aaaaaaaatt ctggtggtcc tggggctcac attcaaagaa agaaactggg gtggaggggt
24721 gttcccattg tggctcagtg gtaacgaact gaactagtat ccataaggat gcgagtttga
24781 tccctggccc tcctcagtgg gttaaggatc tggcgttgcc gtgagctgta gtgtagactg
24841 acagctgcag ctccgattca acccctagcc tgggaacttt ctatatgcca tgggtgtggc
24901 cctaaaaaaa aagacaaaaa aaaaagaaga ataagaagga ggaggaggaa acacggatga
24961 atacatgcac aaaagtggga aaggtctcct ctctggactg gacctcaccc ccatttccac
25021 ccccacccca acccccaccc tctgccctag gtggcttata gactggggaa cctggatgtg
25081 cgggcctttc tccggaacag acaagtggaa cgagtgtaag tgggactgcc cccacctggg
25141 ccccggtcct tacctacacc cctcacccct cggcccctca ctgtgcccct ctcacacagt
25201 gaactcccag caataccggc ctcaggaaac ttccttcaat actggaaagt catctcggag
25261 ttccagtacc gccctggggg ccctgtcatt gacttcctga ataccaggct gctggacgca
25321 gtggtggagg cattcttatc ccaggcctca cagaggaggt ggaagagaag tgaggagccc
25381 aggaacaatg tggtcttcca acccatctcc aggagagatg tgctcagtgt gatggctggt
25441 gagtcactat ctcttggaca gcagcaagga tagctgaggg agcgggtaag cagagggagc
25501 ctgccacaga tctcaggccc acacaaaccc atggctgaag tgtattctac agcaggccac
25561 agacaagaag gaagtaaatt taggggggtt ttgtttgtca tttttaagag aaattgggaa
25621 ccttttaccc accagccttc tagattcagg aacaggagca caaagtactg cttgatgaat
25681 ccccacgtgt acagtttcac agagcaactg ctccaactgc tgctttcctg accattgctg
25741 ctaatggttt tctttctata aaattgttaa tgatttggtt aacatttatt ttataaatat
25801 aaacaaggac attgtttttg cctttgcctt cgcctccacc tggagatata aagttgaagg
25861 ctggagacct ggcccagggg cagaactatc acctccttga aagagcgtga ccctgtgcaa
25921 aagtccatcc ccagtcaagg cctcaacacg ctcatctgga aaaggaggtg ggatgtctga
25981 ggtccctttc agctctgaca gactctaatt atcccagaag gggaatgagg taggttggca
26041 aacttgccag cgaaggaaag gaatagccca ccctgtctta tgagtggggc agagaacagg
26101 acagtggcca tgccaaatgc cacagaaggt gctgtgttac ctgagcctag ggccaagcaa
26161 gagcttcttc tagaatccag gctaagctcc ctcccttcca ggggcaacaa tgagaaatcc
26221 tccaaactac tatggtatgt gtggtaggga ggcagtgttg gggataaggg aataaatact
26281 ccagacttga ctgtgccttt tcttagagaa aacaaatcaa caaaaccccc acaattttta
26341 ttatgcaagt acaatatgtt ttgtgtttag gacacagaag ttaacagaaa aaaaatcacc
26401 tgtattctaa tccccagaga caaatcccct ttgcacatac tctgctaaat tctcttttat
26461 gcataggaca cttcatatat caataaaggg atcaaatcat acctaatgtt tgataacctc
26521 tccattactt taactgtgaa catattacct tgtacattgt attcacattc tgatttgata
26581 tattatttta tgtcttaaat agtgtgtttt aatatatttt aaatagaatg tttttaaata
26641 tttttaaata ttttaaatag aatagtgact gttaaactga aggcttacta tgtcaggcac
26701 tgtgctaaat actgtgcata tattacctca tttaaacttt acagtaaggt agtttccttt
26761 taaaaataag gaaggaagtt cccgttgtgg ctcagcagta acaagcccaa ctagtatcca
26821 tgaggatgca ggttgaattc ctggcctcca tcagtgggttaaggatctgg tgttgccatg
26881 agctgtggtg taggtcgcag acacgtttgg atcctgtgtt gctgtggctg tggctgtagc
26941 atagactgcc agctatagct cctattcaac ccctagccag ggaacttcca tatgccacag
27001 gtgtgaccct aaaaagcaaa ataaataaat aaaataaaag gaaatataaa ataaaaaata
27061 taaggaaact tataaaataa cgtaaaatataaaataaaga aataaagaaa ctgaaggaat
27121 tctcttgtgg cacagcaggt gaaggatcca gtgtagtcac tgtagaggct taggttgctg
27181 ctgtggtgca ggttcaatcc ctggcctagg aacttccaca tgctataggc agaccaaaat
27241 ttaaacaaaa aaattttttt ttctttttgc catttgggcc gctccctcgg catatgtagg
27301 ttcccaggct aggggttgaa tcagagctgt aactgccggc ctacaccaga gccacagcaa
27361 tgcaggatcc gagccgtgtc tgcaatctac accacagctc acagcaatgc cgaatcctta
27421 acccactgag caaggccagg gatcgaaccc gaaaccccat cgttcccagt cggattcatt
27481 aaccactgag ccaccacagg aactccaata aagattttta ataaaaaaat aaggaaattg
27541 aggctgaaag agttaagaaa cttccccaaa ataacacaac tagttagtag tagagtttga
27601 atttcaactc aggtctagta gagttcaaag catatgctct taaccaatgc acttttcata
27661 ctgtggtcct catccattgc acaatattct ttatttaaca aaaccacttc tgggagctct
27721 cgttgtggct cagcgtgtta agaacctgac tagtatccaa aaggtttcgg ggtctatccc
27781 tggcctcgcc cagtgggttg gggatccagt attgcagcaa gctgtggtat aggttgcaga
27841 tgcagctcag atctggcatt gctgtggctg tggctgtggc tttggctggc agctgcagct
27901 ccaattcaac ccctagcctg ggaacttcca tatgccacta gtgtggccct aaaaaacaaa
27961 caaagccact tctgcttatt gttgctctta taaaccatgc tgctacagat ttccgtgaat
28021 atatgtagtt tttgtgtgtt tccaactatt tctcaggatg agtgcctaaa atggaattct
28081 tgacagaaaa ggtatagatt tggtgtaaat ccattggttt ccattttgct tgtgtcctat
28141 ctgacagtga atgtgagcac actgaaaaat tacttccaat gtaacggcta caaggactac
28201 aacctggttt tcagcccgga gagaggcttc acctgtgtgt ccccgtgcag caggggctac
28261 tgtgagaatg gaggccagtg ccagcaccta cccgaggggc cccaatgcag gtgagcaggg
28321 tgggggcagg aggtggaggg cagtgccaga gaatccaagctgcaaggagt ctggaaatgc
28381 catggtacct cacagcgcct gaaaatgtag ggtggttccc tctgtaatcg agagaaaggg
28441 gatcagagaa aaagggacag aggaaaagaa gatatgagtg cctacatttg tggagaataa
28501 agatgctata taacctttgc acctcaaacc agcaaatgga tagatagata tgaccatttt
28561 aaaacgcaac ttagaaatga taaggaatct gactacggta cccagtaatt taagttacag
28621 gatagtgggt tccaaagcca aactcttcca cctgattttg cttctcgggt catggacagc
28681 ctactatctc cttgtggaaa aggaacccac tgtcctgtat ctaggccttg tcctggccca
28741 ccttaagatt cccagaaagg ggaggccctc tcaaactggt gccctctgtc cccttggttc
28801 atcaactggt ctgaggggct ccggtgagtg gagggcacta agttctgggc atgctaagga
28861 gaagcgcagt gcctcctggc gctcttgtca catcggtgcc ctccgtctac catggccct
28921 actacagtcg cagatgggct cagccctctc acctcccagg tctaagaagg cagcgggtgg
28981 agcacagcgt tgggggctgg gtgcagcttt ctgggagcta ggtgtgctca cctgcggcca
29041 actccgcctc cacctccacc ccccatccat gccctgctgt aggggtggag ctgaccccaa
29101 gtgaattcac taccagccca cctaggttgc tcagtggcct cggtttagca ccgcttgctg
29161 cctgcctgcc tgcaggggga ctgaccccat cccttcccca gagagggacc cagcctgttg
29221 tagaaaaatc tcctctggct gggctgagag caggggtggg gaggctaagg gccagcctcc
29281 ccatccccac accccgctgc tgaccggtcc tctctctctc tgtctccagc tgtgtgccct
29341 tttcatccct acgccctggg agcagcgct；
(3)多态性位点的判型:
4个多态位点:g.13383C>T，g.15580A>G，g.21208A>G和g.26002A>C均采用PCR-RFLP(Restriction fragment length polymorphism，限制性片段长度多态性，RFLP)技术进行基因型检测，这4个多态位点扩增引物分别为SNP 13383 FP/RP；SNP 15580 FP/RP；SNP21208 FP/RP和SNP26002FP/RP(详见表1)；聚合酶链式反应(PCR)反应体系为25μL，其中包括40ng猪基因组DNA，1.0mM MgCl2，100mM dNTP，正反向引物各10pmol，2单位DNA聚合酶(Taq酶)及1×PCR buffer(缓冲液)(上海博采公司)；扩增条件为:95℃ 3min；94℃ 30s，56-61℃ 30s，72℃ 45s，共35个循环；最后在72℃延伸10min。
PCR-RFLP中涉及的限制性内切酶详细情况见表1，限制性内切酶均购自NEB公司(英国)。酶切反应体系为10μl，其中包含5μl PCR产物，限制内切酶0.1μl(20U/μl)，10×缓冲液1μl。酶切反应温度均为37℃，酶切反应时间为4h。酶切反应后向每管产物中加入2μL内切酶试剂盒附带的10×loading buffer(上样缓冲液)，混匀，经2.5％琼脂糖凝胶电泳，溴化乙锭(EB)染色后，于凝胶成像系统中观测。
具体判型方法如附图2所示。
通过公用的2LD软件计算4个多态位点之间的标准连锁不平衡程度(D’值)，这些位点之间的D’值处于0.94-1.00之间，由此表明这4个位点紧密连锁，处于同一连锁相。然后采用公用HAPLORE软件构建单倍型。
本发明的第二个目的是这样实现的:本发明利用这4个多态位点及侧翼DNA序列信息，采用分子生物技术鉴别个体基因型，然后构建单倍型，针对抗腹泻病性状选育种猪。
本发明鉴别了一个影响断奶仔猪抗腹泻病性状的MUC4分子标记-单倍型(haplotype)，通过现代分子生物学技术检测该单倍型，利用标记辅助选择(MAS)选择有利的单倍型对个体留种，可以显著提高种群断奶仔猪的抗腹泻病能力，大大减少仔猪死亡率。
附图说明
图1为4个多态位点的PCR-RFLP判型图，其中(a)g.13383C>T位点:泳道13为50bp分子标记；泳道1、5、6和8为CT型；泳道2、3、4、9、10、11和12为CC型；泳道7为TT型；(b)g.15580A>G位点:泳道13为50bp分子标记；泳道2和3为AA型；泳道1、4和9为AG型；泳道5、6、7、8、10、11和12为GG型。(c)g.21208A>G位点:泳道13为50bp分子标记；泳道5为AA型；泳道3、6和7为GG型；泳道1、2、4、5、8、9、10和11为AG型；(d)g.26002A>C位点:泳道13为50bp分子标记；泳道1、2、5和6为AA型；泳道7、8和12为CC型；泳道3、4、9、10和11为AC型。
图2为ETEC F4ac与刷状缘的黏附检测结果，(a)为不黏附型，(b)为黏附型。
具体实施方式
下面结合实施例并对照附图对本发明作进一步详细说明。
选择种猪核心群个体，利用上述的PCR-RFLP方法鉴别MUC4四个多态位点基因型，构建单倍型，选择有利的单倍型对个体留种，群体继代选育后改良种群的抗仔猪腹泻病性能。
实施例:
利用自身构建的、以白色杜洛克和中国二花脸猪为祖代亲本、用于功能基因定位的猪资源家系为实验动物。347头资源家系第二代(F2代)以及108头第三代(F3代)公猪个体饲养到240日龄屠宰，取小肠空肠部分，提取小肠上皮细胞刷状缘，利用E.coli F4ac菌株黏附猪小肠上皮细胞刷状缘，通过相差显微镜检测细菌的黏附结果。每个样品检测20个以上的刷状缘，若有10％以上的刷状缘上至少有2个细菌黏附，判该样品为黏附型，若至少有2个细菌黏附的刷状缘不到10％，但有1-2个细菌黏附的刷状缘多于10％，判该样品为弱黏附型，否则判为不黏附型。黏附效果图如图2所示。
采用上述PCR-RFLP方法对347头F2代个体、108头F3个体以及所有的F0(祖代)和F1(第一代)个体开展g.13383C>T，g.15580A>G，g.21208A>G和g.26002A>C等4个多态位点基因型判定。然后采用公用的HAPLORE软件对这4个多态位点进行单倍型构建，结果仅存在3种单倍型(C-A-A-A，T-G-A-A和C-G-G-C)。采用传递不平衡检测(transmission disequilibrium test，TDT)分析了这3种单倍型与ETEC F4ac黏附表型之间的相关性，结果显示这3种单倍型均与F4ac黏附表型呈极显著相关(见表2)，单倍型C-A-A-A优先传递给黏附个体(即易感性个体)(P＝1.90E-13)，而单倍型T-G-A-A优先传递给不黏附个体(即抗性个体)(P值＝6.52E-8)。单倍型对在黏附表型中的分布见表3，几乎所有携带单倍型C-A-A-A的个体均为黏附型，即单倍型C-A-A-A为易感单倍型，而几乎全部单倍型T-G-A-A纯合子均为抗性，即单倍型T-G-A-A为抗性单倍型。单倍型对T-G-A-A个体是抗腹泻病型猪的选育改良所需的单倍型个体。在商业种猪核心群中，继代选育单倍型对T-G-A-A个体，可以进一步改良种群的抗腹泻病性状，减少仔猪的死亡率。
表2 MUC4 单倍型与F2个体ETEC F4ac黏附表型的相关性分析(TDT分析)
表3 单倍型对在资源群体不同黏附表型F2/F3代个体中的分布
Claims (2)
1.一个可鉴别西方猪种断奶仔猪F4腹泻易感性/抗性的MUC4分子标记，其特征在于:该分子标记的多态位点序列如下:
1 gctactcatg gtacttctga ttggggttcc tctgataatc aggatgttga ccatatattc
61 atgagctggt tgtccatctg tatctctttc ttgaggaaat gtctattcag gtcctttgcc
121 catttttcac ttgggttgtt ggctgttgag ttgtaaaagt tgcttatata ttctagagat
181 taagccctta tcagctgcat catttcaaac tattttctcc cattctgtaa gttgcctttt
241 tgtttatttt tggtttcctg tgctgtgcaa aagcttgtca gttcgattag gtgtcattgg
301 tttacttttg cttttatttc tgttgctttc ggaggctgac ctgagaaaac atttgtaagg
361 ttgatgtcag agaatgtttt gcctatgtta tcttccagga gtttgattgt gtcttgtctt
421 atacttaact ctttaagcca ttttgagttt attttcatga tttgcatgca gctgtccaca
481 tttcccagca attcctgctt aataaaaatt tgtgtatgta ttgcttgaaa ggacagtctg
541 tgtattttaa ttaaaaaact gttttgagac tgaagctctt gtatcttgcc ttccatatat
601 ggcttatatg gagagagtag gagtgttctg ggtccatttc cattatacca cacaatgaat
661 gacactgaaa acatattagc acaaagatat tcactgaagt attatttatg atgaggttac
721 acagtggccg tagaaatatt agacaggggt aatcctgcag ccacttattc tgacccacca
781 tgatatacag tggtacacgc gggaaaaaat tgtgttcttt tttttgtttg ttttgttttg
841 tttttttttt gtttgtttgt ttgtttttgt ctntttgcca tttcttgggc cactcccatg
901 gcatatggag gttcccgggc taggggtcta accggagctg tagctgccag cctacaccag
961 agccacagca acgcgggatc caagccgagt ctgcaaccta caccacagct cacggcaacg
1021 ccagatcctt aacccactga gcaaggccag gcatcgaacc cgcaacctca tgcttcctag
1081 tcggattcgt taagcactgc gccacgatgg gaactccaaa atactgtttt ttttgttttg
1141 ttttgtttat ttgtttttgt ttttgttttt ttccttttct agggccgctc cctcggcata
1201 tggagtttcc caggttagaa gtcaaattgg agctgcagct gccggcctac accagagcca
1261 cagcaacatg ggatctgagc cacatctacg acctacacca cagctcacgg caacgccgga
1321 tccttaaccc actgagcgag gccagggatc gaacccacaa cctcatggtt cctagtcagg
1381 ttcattaacc actgtgccac agtgggaact ccaaaattgt gttcttaatg ctacaacttc
1441 acagaaggaa cctgtcttgt caacgagtcc ttgtatctta tatttaacgg cactattgaa
1501 acatatgcat catcatatct ttaatcctct acaggtggaa acactatgac aactactatt
1561 actgaggaga aatacccatc gaatatcagc tacataactc ccacaaaaga aaacctctca
1621 aaagaagtaa catcaacaaa caaaactgaa agaaaaaaga gggaaacttc aaattctggt
1681 acttacaaat caactatggt gagttcatca gtctcagcga caactggacc agggccccag
1741 tcaacctcac cctcttccac cacctctgct gaggggacag cagcatcttc tctggtccac
1801 cagactcaga gcatggagac caccagagaa acccacagca gcgccatcgc agctgtgagc
1861 tcctcgacac ctttatcttc cacaagtgga caccctctta cagaaggcgc ttcccgggag
1921 acatcccctt cgggggaccc aacctcttca tccgcgtcca gacccacccc cacacctgca
1981 acaacatcag ccgtgtcgac agcacctgct tctacagatg gcacttcccc cacttccagc
2041 ctaagcaaca cacccccaac aacatcacgg gtggtcacat ccccagttac aaccgacacc
2101 actgtgggag gcccagggga cacgtcccca cccggcacaa gaaccatcac tccaggcacc
2161 tcatcagtct caacgacaac tggaccaggg ccccagtcaa cctcaccctc ttccaccacc
2221 tctgctgagg ggacagcagc atcttctctg gtccaccaga ctcagagcat ggagaccacc
2281 agagaaaccc acagcagcgc catcgcagct gtgagctcct cgacaccttt atcttccaca
2341 agtggacacc ctcttacaga aggcgcttcc cgggagacat ccccttcagg tgaaacaagc
2401 tcttcagcca catttagacc gacccccata tctgtgtcgt gtttccatac ataaaccagt
2461 atttcttctt tttctgcatc agctcctggc cacaaagtca ccttgagctc cactgcccta
2521 ttcacaggtc accccacttc tctgtctctc atgagccctt ttcccttatc cactgtgtct
2581 tcaagatcca cagtgaaggc tgggacatca ggtaggtcgt aaccctgggg tcttagtctc
2641 ttgcacttgc ccagttggct caattcagtg cagaagagtg cttgagagag gacattcaat
2701 tagcactgcc aggctggagg tcaggtccag aaggttatga aatcagttgg ggtttggtgg
2761 gtgatagcct caccatcctc tttaccccag gtggcagctg cttctcctca ggcagtggtt
2821 ttggtgtgac tagaagacag ttttctagct gtctcttcca ggttctaaag tctatgactc
2881 tgtttaactc tgctaccccc tgactgttag actcactcct gtcttctctc cctgcctgcc
2941 taggaacaat caccaccttg cagaagacag acagcaagac aagcacagca gcatctctac
3001 cctcaaccac ctcccaggct cttacaactt ccactgctca cacttccaca ggcactcgct
3061 caacagctgc cccagtccct cccaagccga agaaaggtga gtgaatccat agagctgggt
3121 cctcttcacc ctgggtcaaa gaggatggag ctatatgtgg aggttatagt tgtggcagct
3181 ggactggatc taccaatgag gatctttgtg gggaggtagg gggaccagtt ctgtggatcg
3241 ctccactgcc ctgggtgtgg caagtccagc tgctctctgt gtccccttga taggggtggt
3301 gtaactgagt gctttgtctc agcctgcttg cttacagact ctcgagtcag aaaggcctta
3361 cagtgaacaa tgtgacatca tcagtgttgg gtgctgaggg ctgactttgc acatggtgct
3421 aggctaagag ctggagataa aaaaatgatt gagtcactgt ttagtgagac agccttgctg
3481 cctcagtagg tgcgattaga ggcataaacc cagggtatta ggtacacaca atggaaggcc
3541 ccaagggaat attggggtgt gggttcaagg tgaaaaggag tctccctgga aagggtgaat
3601 aggatctgcg gggaggggaa gtgccccaag caggcaggag ggcctgaata gctagaataa
3661 aggctcggac aggaagagca gtcctagtat tgcatgggac agagatcccc atagcatttg
3721 gggctgccag cagtaaggtg aaaggtgttg agaagcaagg ttgttatgtg aaggaatctt
3781 gtagatgaca ggaaccaggg agaatgtggc tagatttgca tttaggaaca gtgtcctcat
3841 ggctgggatg gaggatgaac ttgcaggagg ccagggcagc agggtagata tcactgaggt
3901 tttccaagca agaccaggtg aaggctgcca gtatggaaag gaggaggccc ctttgagaaa
3961 tgtttgggac aaagtttctc aaaccggagc ccaagggcct cggtagggga tcttatgaca
4021 aattctagca gggggcagtg tgtgtgtgtg gcgggggcgg tcttcaagtt tacctagaga
4081 cataacatct taggaaacaa gtctcattct gtgaaacgat cttgcaatta ctttactgca
4141 ttagaggcat tttgtctttt aacatttcac agaattgtga gtaccgtaat aggtgcagaa
4201 taatgtgggg gtgttctgat agctgttttt cttcaaactg gggaaggcgg agagtaccat
4261 gtaagggagt gaaagaggag atgggttaaa tgcaagtgga gagggggaga gagaagtcct
4321 tgacggctct tgcttaagcg aacaggtcca agggtggagg agcagcttgg ggacactctt
4381 gcgggacccc tgccagccct aatgacatca gggcccatga gaacttcctg cttctctttg
4441 tactggccca ggcgttcccc tcttccccta cgggccgcag gctggagact ggcagtttgt
4501 gagaaggacg gtggacttca cctcgccact cttcaagccc cagattggct tccccctcgg
4561 ttcctctctc cgggattgcc tctacgtgag tccttggctg cagcccgaag gcggggggcg
4621 gggggcatga cctgcattcc tcaggccttt cttctccctc cagggctcag agatggccct
4681 gtgatcatgg ccataactgt gactgtgatt gacagggaga acggaagcaa cgcttgtcag
4741 gcacctgctg tgccccagtt gctgcctcag tttgcaccat ctttaacgat gggcaccagc
4801 tgaggaagac agggagctag gcgaggcccc ttacccacgt ggacagaagt gtggccacct
4861 cagtagagag gtcctggggc ccagtaagct ctccactgtt ttcatgggtc cttttttaca
4921 agtaggaagc tctgctgggg aaacctgtat gatgctgttc acacggtctc tcattgtttc
4981 ctggttccct agtccaagta gcccatgtct ctggggggag agcagaacag cagtcgtggc
5041 cctcgagagg gctggtcagg acacactaca aaagggtgac tcagcaatgg ggtcagggta
5101 tcctttagcc ctaattgatg tggcacaagc ttcagggtgg gctgagaggc tttactctgc
5161 tctgggatgg gctggtctct agacgggtgc cactgagaca ggacaaaact accaccttag
5221 gaaagactca ggacataaaa aaaatgtgct gagggtgaag agtatttgtg tgagaactat
5281 atacagaggc tgtcccagca tgcagggaga tggatgagac cttccagagg ttgttggcat
5341 ctctctggtt ctacccagca gggcccttca gggtcttgca tcagtccagg agctggcccc
5401 aagccgaggc tccttcccca cctcccgccc tgaatgctaa ccaggagaac ctttgtgttc
5461 agttcacaga taatggccaa atcattttcc cagagtcaga gtaccagatt ttctcctact
5521 ccaaccctcc cctcagaggc tttacaggcc aggaccctgt ggccctggtg gctccattct
5581 gggatgatgc tgatttctcc aggcaaggaa ccacgtttta ccaggtgggc ctttcaaagc
5641 tcagcattca gggtcctcca gcagccagca aggacagaca gagggctgtg tggagggcct
5701 tgcaagtgtt actgctgtca gagcctaggc tcagtgtggg cagggactaa aacttaaatg
5761 tgaggatgag gaggaagcag atgaggacaa aatcatggga agactgaagg tgccagggat
5821 ggggattggg gatccatcgt ttgattttag gatgaggttg tagcagggtc cctcctaata
5881 aggctgagtt aagcaaggcc cctgcattgg gctgatgatt ccattcagtg ggagctattg
5941 tgtaatgaaa gctaccacag gcaaaggtac gtcgtgcccc tgccaaggac caggcactat
6001 agaaggtcca aagatggaga aacatgtgcc tgtcctcaag gagcctccag tgggacacag
6061 ggcgatacag atgggcacag gtggctgcac aatccaagcc agtctgtgct aagcgaggca
6121 caagttcaag tccagaagtt actggcctcg actctccact ccacataccc caggtttccc
6181 tgcagcccga acctgatttg atcttaaggt cagaggtgct aactgtgaaa atgtgggcct
6241 gggacggggg aggatggaag atatgggcag actcaaatga gcaggatcac catgcagaaa
6301 gtggggggtc gcccaggagt gttgagctgg gatccttgtt tcttgtctga atctgccttc
6361 ttttctagga gtatgagacg ctctatgatg aatacaacca gctagtgtgg gaggtggagt
6421 attggattaa aatgttaaca aacacctgga actacaaagc caggtggaca ctaaaggtca
6481 cgtgggtcca cgcccctgcc tatcctgccc agaagaccct cagggtgagt ggaccagtgg
6541 gcagctccct gggaaccatc tggaagtcag acatcctaga atcctaggca ggggccactc
6601 ttctaaaatc ctttcggact cttacagaac ctcacactag cccagagcac tgggtattag
6661 gaggttgaaa catagaacgt ggaatgtcca taagtcaaaa atggctggca attggcaatt
6721 ctatatggct caacctaata tttttatcca catattagaa acgaggagtg ccctggtggc
6781 tcagcaggtt aaggatctgg cattgtcact gctgtggctc tggttgctgc cgtggcacag
6841 gttcgatccc tgatctgaca attttctgaa tgccaagggt acaacctaaa taaataaata
6901 gaaatagaaa gaaaatagcc agaaaatata actgtgaatc aggtagtcaa aatacatatt
6961 tttttaaaag aacgaaatga ggaagatgat ggttagaaaa gttaagtgac ttgcttcaag
7021 gtcacccagc tagcaaatga caaagccagg cctgaaacag aggtttttct gactctgcac
7081 tccagctctt gaccaaaacg catccactgg aagcaggcac accacatgtc cccccctatc
7141 tggggcctca ccaccctgat agtcacaaag ttcttccttg tgtctggtct aaagtcactc
7201 atttcttcat tggctcaaca agctcattgc ttacctactt tgcacaagca cagtgctggc
7261 cccggccttc agagaacctc cactctcacc tgtgtggtcc tccacgccat ccaggtggag
7321 gggacatttg gacaatatcc ttaaatatcc ctgaggaagt tccttaggcc tcccccagct
7381 ctccaacgtc cgctgatcac tcaggttgct gaagctttct catacaaacc ctcttctcac
7441 tgaattctgg cctagagtcc tttggctccc ttcctgggga agaagtccca aagaaggcaa
7501 atccaccatg gcaggtgctt ggacagtcag agtgaaatct ctgcataatt ccaaatttgc
7561 ccatatttta agcccacacc ttagctctgg gtaggccagg aaacagtgat atgggtgtca
7621 tgcccaggct gtgtgacagg taatgggaca taccagatct ctcacgggca tccctatgct
7681 tgtctccaac caatccccag accaacacct accaggccat cctttccacg gatgggagca
7741 ggtcctacgc cttgtttctc taccaaagcg gtggcatgca gtgggatgtg acccaacacc
7801 taggcaaccc cgtcgtcatg ggcttctcca ggtaggacag agtggggctg tctgcattga
7861 gtagacagca ggaggcggtg caaagcaggg cttgcagctg cacacgaact ccattcccca
7921 aggatacgcc aaaatcctcc ctgcccatgc ttctgattca cgctgactcc agttcccact
7981 tcttgctgtt tcaaattaaa agacccctat tcactctgcc gttctctttc ctatcaaggt
8041 ctccgtgcat gtgataataa tccccagagc cttgacttct aggacttcca ctgacagtag
8101 ctgacctcac ttgacttttc agagcttaac aataacagcc aacatttatt ataccagcac
8161 tcagtatgca ttattttgta tattacaaca accccatgaa ggagatctat tttattaccc
8221 attctacaga tacagaaact gaggtacaaa tagataagtc attgcccaaa gtcacagagc
8281 tgggatgtgg caaagtggga agctgaaccc aaatataagg ttgaaatctg ccttcctgac
8341 tttcgattgg ctggaaatta acctctcacc caaggcacca gcagcgtctg tcagtggcaa
8401 agatggagaa cagagagatg gatggggacg tgggttccca agggaggagg tggctcagtc
8461 tctgacacaa cagttctcaa ccttgccatg tgcgccactg gctgtgaagg tcgcatgtaa
8521 tttgttactg ctaattgcta caaatgtttt tctcttagag tgtattcagg attgcgcctg
8581 taattacatt gctttcactt gcttagtgat gagccctgaa ggaaaaagaa gtagaattat
8641 ccgaccagct aggaatttca caccattctg agtgcaccca cggaaggtgt cacacccaag
8701 tgcgtcatct gtcctgtgtc agagaggaga aaagatcagg agtggctgct ctgaggcgtg
8761 caccctgagg ctgtgggctt tgctccttcc ttccaggggt gggggtgggg cacacccatt
8821 gcctggtcct cactcggtgc tgagggattt accagtgtta ctgctttgaa tcttcacaat
8881 actgtcaccc ctttgacaca gaggtgaaaa ctaagaatca aagaggttaa attaacttgc
8941 ccatgatcac caagctacta agtagtggaa ctgggggttg aatccatgac cgagagactg
9001 gcttttaacc ctcacagcca gctgtgccct gcagcacccc ttctccaatt cctagttgaa
9061 gggaaactaa caggaagggg gcctgacctg ttgttctcac acccctgtct catgacccgt
9121 ttcctacagt tcaccatcag atgtggaatc aattagagtc tcctattcct gccccaagcc
9181 tcccttcccc actcctgctt ggcaccgggt ctatgattct ggctggctga cgaaacgact
9241 gaacagctga taactaatga ctggctggac tgagtgggtg accagctgac tgaccagctg
9301 tatgaatgag taacttgctg actgagagga aaaactgtgt gacagtggct gaagattctt
9361 ccttttgcct acagtggaga tgggtatttc aaaaacagcc aactgacatt gcagccagtg
9421 tgggagaagt atcgtccaga ccagttcctg gatcccacat caggtaaaag caccacctta
9481 tcacacttga gggacccaag tccccttctg tcctcccctc catgtgccat gacagtccca
9541 gagggcagct cccttgcatc tccaccatcc caaagggtgt ttccagcaca actgacttcc
9601 catggtgcaa tccagatgtg gcactgccca gggagtggtt acctcccgtg ccctcacaca
9661 gcacaaagtc atttgaggct gcttctcctc cctgggcgga gccaaacaca ggagagctat
9721 gagtgggggc cctgatcctg gaagtgagga caggggcatg gcaggaggtg aggaatgaat
9781 gggcagctag gacaattctc caagtgtcct gtcccagaac agtaatgaga cttcattctt
9841 caatggaatc taccaagagg agctccaggg gatctgacat ctattcagtc ttcgtttatt
9901 gagtacctgc tgggtgtagg acactgtgct aagagccaga agtatgagaa atgaaaaaga
9961 ctatgtttgc tgtcaaagag ctccccctct ggtaagaagg ccagtactct aggccccacg
10021 ggcctcctcc atgatttgat tactgtccgg tgacccctaa agaaaagaaa tttcagtttc
10081 caaatgtgac agaacaaagg atattctggc attaaaaatc tgccatgtaa agtatgaagc
10141 attaatccag ttctgattgc tctgtggcaa caacttctga taaacagaag ttaagtcaaa
10201 tcatgggtgc catcttgtgg agaaagctgt cctattagac tagggccctt ctatccagtt
10261 tgtgcttgta gggggcgcaa taggatagtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
10321 cgtgtgcatg agcgtgcatg taaggcccct ctatccagat gcatctgctt gtgacgtggg
10381 gatgcaatag gagtgtgtgt gtgtgtgtgt gtgtgtgttg ggggagagta tctggtgtgt
10441 ttcccctgat accaggccac agtggagaca atgcacagat gatcataaac aatgattact
10501 cttttccaaa cctcatcaaa actgtagtta gcttgatact ttcttttaca cggggtcagg
10561 gtcaaaatca aatacattat atcctattgt attttactgg actctggagc ccacactcag
10621 ggtttggctc tgagttggat tacaggtgag gccctcctgc ccagcacttg tctcaaggtt
10681 tcgctgtaag atccaaactc ttgaggtttt ggaggtcaca tcaaagggcc tttggggagg
10741 cactactgcc agtctgactg ggtccattcc tagaagagct gtccagggca gggggagtga
10801 catacagagg gtgcatttat gagttttgtt tgttcccaaa actaaagcag agcctgtggc
10861 ccaggccctg agatcacccc cttttcctct gctccccagg cctccggggg ctgcagatct
10921 ataagctaca caaggaagac aagcccaact atcgtctctg gtgcctgcgg tggttgaaga
10981 gacagtctca ctggtccggc tggggctgga accaggtctc ctgcccctgc tcctggcagc
11041 agggactatg ggacttaaga tttcagccca tcaacatagg tgacacctcc ttcatagtat
11101 cctgtggtgc cccactgtcc ctcacaagcc tgcccaccct ccctttggcc ttgcctgacc
11161 cgtctcttcc agggcaccct cagttcctcc ccagaagcac cccctaccgt tgatgcacac
11221 accaaaggca tcctgggcct gagtgtgtct ggcttgttgc aggctggtgg ggcatcgtca
11281 acaggcagct gtgcagcttc tcctcctggc gtgggagtgt gtgctgcagc tacgggccct
11341 ggggagagct tcttgaaggc tggagattgc agagtccttg gcagtttggt atgtgtgtcc
11401 acagagaact tgagcccagc ttttctggag ctttctgttc ttccctgtca gagaccagcc
11461 agcagtgaga cccacagaag gaaggctctt gtcccatttc ctcccagcta ccttccccct
11521 ctggccttca atttcctgat ctggtaacat gaggaaattt ccaacatccc ttccagctga
11581 caccctaggg atttatgatg gagtccctct tgtcccatct ccacttcgcc aaaagcccca
11641 gcccagagtc gggcatcttc tagggaagag aggggcctgg tctggccctg tccatcggaa
11701 cccagattag cagctgtgca ggggaagact tggagtcgcc tcctgcagag accctccaaa
11761 gcagctagag aggacagcgt tggggggcca gggtcatgcg tgtccttctc acctcctcct
11821 tgtaatcaca gcctccttag tccccacacc accagccatg ccctcctgct cccacagcac
11881 tttgagactc agcaccccac ccttcaagaa aggaagggcc ctgggagttc ccactgtggt
11941 tcagtggtaa tgaacctgac tagtatccat gaggacacag gttcaatccc tggcctcact
12001 cagtgggtta acaatctggt gttgctgtaa gctgtggtgt aggtcgcaga cacgacttgg
12061 atctggtgtt gctgtggctg tggtgtagac ctgcagctgc agctccgatt tgacccccag
12121 cctgggaacc tctatatgcc gcaggtgcgg ctctaaaaag acaacaacaa cgacaacaaa
12181 gaaagaaaga aaggaagggt cctgtgcctg gcccactagg ggctttgcta cacatggctc
12241 caaataccca caagggagac agtcactggt caatggcttc aggtcattga gcagggtggg
12301 ttctgaactg aggtgtgccc gattcctaaa ccataccctt gccaccgttt catcctgcca
12361 gaactgccca gtgctacttc ctgcaggttc tccatctccc ccaacacgct gctcttcttc
12421 acagaccaag aactggagcc ccagaactgg tgctgccgct tcaatgacaa gccctccttc
12481 tgtgccctgt atgagctaat gcggccccga atcggctgca gtgggtacca gcccccgagg
12541 cctggtgagc tgctggagcc tggctccagg acagggcaga gacagagcat caggggaagg
12601 gtagcctctc aggacctgac caaggcagca caagcttcct ggggcagggc tgtggccaca
12661 cgggaggatg gagatgggtc cagtgtgtgg agatggcaaa cgagaaattt tatatgtggc
12721 cttgggggag gggaggtgtt gggggtggga aaatgaagat gcggtggggc gcccaagggc
12781 aggaggaatt ctggggaggg gtctgtgctc catgagcacc tccccaggct ggagtctcct
12841 gagcaaacat cctgtctggg gccagtaagg aatcccaggc ccttccctgg tccactctgc
12901 caatgtggta tctcctcttt tccagcctgg atgtttgggg acccccacat caccaccttg
12961 gatggtgcca attacacctt caatgggctg ggggacttcc tgctggtccg ggcccaggac
13021 agaaactcct ccttccagct gcagggtcga actgcccaga ccctctcagc ccgggccacc
13081 aacttcattg cctttgctgc atattacagc tccagcagcc tggaccccat cacggtgagc
13141 cttcagctgg caattgagtc ccaggaggag aaagagaagg aagaagggca gggggcagag
13201 gtgagggagg atgcatcctg ctggcacagc agttccagag tcaaccctgg aaacatagga
13261 ttagggtctt ggctccacca gctgtgagac tgaagaaagg acctactttt cagtgcctcc
13321 cagtttttct tagggtaaaa tggggacgat actactacct acacacgggg ctgttgtgag
13381 atctaaaagc taatctaaga acccgtgcca agggcctctt acacagtggc tagcatcacc
13441 tcatcgtcac actcccagtc accttcgctt tcatccaagc cccaggctct aagctctcgg
13501 aagccttctt ccttggcctc tggacccacc tgcaggtcgg gcttaggcct ctgccaattc
13561 ccttccaggt tcaatggctc cttgagccca atgacacaat ccaggttctg ctcaataacc
13621 agactgtaac atttgagact aaccatgcag gtgccgaagg taggctgggg atggccgtca
13681 gctgccctct cctgcttccc tgggtcgggt cactggggag cagggtgctg gggtggagag
13741 gcagcagcat cgggcagagc agagtggtcg ccacccctgc tgtccttcca gcagctgtgc
13801 ctgagcagct tcttcactgg agccgagcat tgagcttttc ctccgcactt ctctccttcc
13861 tccagggctg tgacggcctc tctgggcaat gccagggaga actggttggg ggggcgggtg
13921 ccatgagggt gtcctgtgag cttggtttgc ggctactggt gtaggacctg gcagggcatc
13981 tcccagctgc tctcagcctc ctctccacat cttgaacctt cactactgcc aagtcttggg
14041 cctctaggac tccaacactg gctgtttcat catctgtctc cccaccttct aggccaggag
14101 atgttcaaca cctctggagt cataatgatc cgcaatggct ccacggtgtc agccagcttt
14161 gatggggcgg tgaccatctc agtgatcgct ctcgcccaaa tcctccatgc ctcctgcggc
14221 ctcccaaagg agtatcagaa ccacacagag ggcctcatgg gtaaggagcg ggcaggcccc
14281 tgcctctgca gcaccagctc agacccccgg cctacaggca gctccccctc ccgtgggtct
14341 gcggatactc cgggtggagt ttcctttttc ggcctcctgg gagcagagcc ctgagggtac
14461 tctgaagctc tggttccagg gcagccaaga aggaatcaat agtgggctct gggagttccc
14521 attgtggctc agcagtttaa ggacccagtg ttgtctttgt gagaatgcgg gtttgaaccc
14581 tggcctcact cagtgagtta aggactggca ttgccacaag cagctgcagc gcctattcta
14641 ttcaacccct agcctgggaa cttccatata ctgcaggtgt ggccataaaa agagaaaaaa
14701 aaaagaatgg gctctgactg ccttgggtgg cggggggggg gggaggaaag gggactgatg
14761 gagagtttga gttggtaggt gtaaactatc acattcagaa tggataagcc ggggggtcct
14821 tttttccccc cccgggaact atatcccatt ttttggggta gaccatgatg gaagattttt
14881 ttagaaaaaa ctttttttaa gtaagggggg ggggggccct tttggggggc cgccgaaatt
14941 gccccccccc cgtaaatcaa ctatatgcta attttaaaaa aaaaaaaaaa aagaatgggc
15001 tctgggaatc atgggaccag cagccacccc accccccact ctggctttat acctcccacc
15061 ctccctccat cctccgcatt ccctcccttc tttctccttt cctccttcat ttccttcttt
15121 cctgaagcgg aggggtgggt tctgaggtga cagcccgttc ccctcctcct ggaggactga
15181 atggagaaga gatacaatgg acggaggccc cctactcctg cacgcatctc catgtgacca
15241 gccggatgct cccagggaag ggcaaagcca cagtcccagg atggctgact cctgctctgt
15301 ccctcagggt tctggaacgg caacccagat gatgacttca ggatgcccaa tggctctact
15361 ctttccccac ggagctccga ggagaccctt tttcagtatg gaatgacctg tgagtctggt
15421 cctcagagtc ctcgggtaga tgggcagggc tgctgcatgg tctccctcag gcctgtggga
15481 accagacctc tgtcctttgt gagagagggg tagaaaggaa gagatctaaa gatgctggtg
15541 ctacccccag atttgtcccc tctgtcccca acccacctgc gcctctctac ctgggagagg
15601 ggacaaggga ggtgggtggc cctcagtcac tagagtgagg atttccagac tcagaagctg
15661 gaagtcactc tgtctctcca catcccctgc cccaacccca aaccatataa gggtaagggg
15721 tggacacaac agaaatacag ccagaggccc accctggtaa caccaccatc atgccttgtg
15781 tgttttaggg gagatcaacg gaaccagcct ccttggcaag aggaatgacc atgtgtctta
15841 taacttcacc cctgtcttcc tttcacaact tcggggaaac aagtccttga ataaaagttt
15901 gacttccagg tgtaatggag atgaacaatg catctatgat gccctggcca cagaaaatgc
15961 aaaccttgga gagcacacta tgcggctctt tcgaagctac cagcaaatga atgctacctt
16021 gagtgagtgg cgtgaggctc ggggaggtgt gtgcagagtt agggggcaga tgaggagccc
16081 ctcttccaag acacccctta agctacacat ttttttccta aagttattca acattttaat
16141 cccctcctaa acataacata aatttttggg gggccacacc catggcatgt ggaagttccc
16201 aagcggggaa tcgaacctgc cccacagcag tgacccaagc tgctgcagtg acaatgcagg
16261 atccttaacc tgttgcacca ccaagagaac tcctaagcta cacatttttt aatttagccg
16321 attattttgc tcatgttgtc cctctgctca aatcctcccc tgactcccca gctcactcta
16381 tccttatgac aaccttcaaa tcaaagccct gtgcactcca gccccactgc ctctccacac
16441 tcttctccta gaacttctag aacctcgctc cctccactgc atccccactg cttcttcatg
16501 ttcttcaaac acgccaggct cgcccccgac tctgagactc tgcacatgtt ctgttctctt
16561 tgcctagaat accttccccc agacacccac aggtcgctcc tcacctcctt caagtctcta
16621 ttaaatgcca ccctctcagc agtgcctttc ttggctaact tgtctaaagt ttcaccttca
16681 ctcacacttc ctatttgcct tccctgattt attttttctc tttagcatct ataactatct
16741 aatgtatttt ctaattgtag caggtctttt tattatttgt tcattgctat gcccttacga
16801 cctagaacaa tgccagccat agagtaggtg ctcaataaat atttgctaag tgactaactg
16861 aatgaatgaa tgaacaacag aatggaaata ccagggctcc aaatccagag accagtggaa
16921 aacttttact ctctcattct acttcctgac aatgtaatat ccacctagat ataaccctca
16981 gtgtttccag ctctcctcca aaggtaaaat tccacaatgg cctgaacagc tctgtcctct
17041 gatcacttca taccatgccc tgcataactt tctcaccagg aaagaacctg aaatgaatct
17101 tgtttgggga gaaaataaag agagagtaaa gatttggctg ggtgggcaat acaacaatga
17161 cctgctctgc atagatgcaa gagcaggtgc atagattatt tcctgcttct ggaggtgggt
17221 cccagccctg ggaatgcctc acacaagcga gaagactaga gagagagaaa ggaggcccag
17281 aaggcaggaa atatcctgcc ccagaccctg accccttctt tgtgtctcag atcagtaccc
17341 gccctccatc aagggtcctg atgtggtgaa aacctatatg ggacaggcca gccttgttaa
17401 ttacaccagc agctctgaga acgtcacatt cactctcaga cacaactgca ctgacttcaa
17461 gctctctggt aagaatgctc ggctggggca gttagtggtt gaggtctgag tcaagctttg
17521 ggtaattttg cgttattata ctctgtgcaa cttcatgtca cctttcattg gaatatctag
17581 gaccaacatt acccaggaat gatccatata aatcaaatag attcccccca gagattaata
17641 gtgatctggc cagctagctc gactgactcc aggctgattc aaagatcatt tcttc tcatt
17701 cttgtaatat tgcaaaatca tctttatgca atcaccaagt tattagaata aatttt tttt
17761 tgtctttttg tctttttagg actgcactgg tggcatatgg aggtttccag gctaggggtc
17821 aaatcagagc tgtagctgcc aacctacacc acagccacag caacatggga tccttaaccc
17881 actgagcaag gccagggatc taacccacaa cctcatggtt cctagttgga ttcgtttcca
17941 ctgcgccaca acgggaactc ctagaataaa tatagcaatt aaaaagtact aatttaaaat
18001 atttgcacat ctattaaaag aaggatatca tatggaacaa gatttaaaga aatacaaata
18061 ccttcatttg gtaactttct cacaattttg atgtttgtca accatgttta cctaactttg
18121 cccctccttg cctgcttttc tgtactgggc ccaagcagtt tcactttccc tctcagtaat
18181 ggtgtggaga ccccatttca gaacgggaac catgatctgg gtcaagagtc aaagcacaca
18241 ggttacagct tgccttttag ggccagccct aattaagctg gtgactgctc tttgctaagc
18301 cagagccctt agggaccttg ctctgagcac ctctctgcct ctctttctct gtgcataaga
18361 agagacagca ttgtctgcac cctgtgctcc atgtggcatg ctccctagtg cctaatcaat
18421 ggcatcagga gactcactcc tccactgggc tcctatcctg aagagggagg tcctggaagt
18481 gactctcatc tgctcccacc catcagctct tttcaatcca ctgttttaag gagacttccc
18541 tattttcttg aaaaacataa gacaggttga atcagaagag tccttattga tacacagcag
18601 tatttccctg tagcctgggg tctggtcaaa ggtccaggaa atgagagcaa gtgggtgagg
18661 gatgaggctg ctgtgaagct gaggtcagtc tggcctgaag tgtagcctga gtaagtgttg
18721 ggaagatcct ggccttcacc ctccatggct ccccgccccc catatcccat cctagtgcag
18781 gtccaggcag ggttaccctt tgggggcttt ggggaggtgg ggggcaacca ccttcatctc
18841 atggagttct accctagaat cacttagttg ggctggtgcc acctccaggt tggagtgtga
18901 tccaacagaa actagaaacc agccttgctc aaggaaatgc ctagtaagat tggaggggtg
18961 gtctgacagg ggtgggggac gtccagcatc agcacttggg gtattagatg ccaggatgtg
19021 tcaggcagca gaggagatga ggctcccagc tctcaaaatg atctgtcttc cctgggggca
19081 gagaatggga cgttgctatg gacaccacag tcactgaaac catgtcctct ggagattctg
19141 gcaagaagtg ccaaggatga cttgtcatct gtactcaagc caagaatggt ggtctgcact
19201 tgccaggcag agagccagtg tttatataac cagaacgatc gggtgggcaa ttcctccctg
19261 gaggtgagtg tgggagaggg tgggaggtcg gtctctgtgt tgagggaagg gaagtgggaa
19321 ttgaagggat gttgtcattc agcccctctg tcctaatgtg tgtgttggga ggtgggtgga
19381 gctgtggtag gtgagtaggg cagataccaa ttttgggcca caaacataag aaataggact
19441 ttctttagtg cttataggtc aactgaattt ttacctccca gctcccatcc agccaagcta
19501 gcaggaaact tccacctcag cctttctttg agttgctcag tggtttctag ctccatggtt
19561 ctggaacgag acaatgaagt gtggggtggg aggaggtcca tgtggcccct ggggggcctg
19621 agtctccatg gatcacagct tcttcatccc atcagatgta gggagatgct ctgtggcttc
19681 tgggccttac ttggatacag gaaacctcct ttaaagtggc ttagggtccc atgtgcctag
19741 aagcagtgca cagccatttc tttggggcct catcgtcatc ttgggaagag aggtacaggc
19801 cccctctgat tttaacacct gtaaaatcct atttttcctt tttgctctct tccaactaaa
19861 cgaagcatct ctttggcagt ttagagaaaa tccttctcca ttcaagtttc tagccaagat
19921 acttggccta agtccatgaa tctggcccta tctgcctttt tttttttaat ggccatgccc
19981 acagtgtatg ggtgtgacca ttaaaaaaaa aaaaaaaaaa acacttctgg gccagggatt
20041 gaatctgagc cacagctccg gcaataccaa atcctttaac tcactgtgcc tgtctggaga
20101 ttgaacccac acctctgcag tgacccaagc tgctgcagcc agattcttat ctcactgcac
20161 tacagcagga actcccccta cctgcttctt aaaaaaaaaa aaaaatgttg ttgttgtgaa
20221 aggggagctg gaaaagacag gttttgactg gcaatactgc accccattct tcaacccctt
20281 acatacacac acaccctcct tatgaaacat gaacacagaa ttctgacttg aaagggacag
20341 aggtggggga gtccaggagg cagaaaaaga aaaaaaaaaa tcagaatgga gaaaggaggg
20401 attaaaatca atacattatt ttggatttcc caagatatcc cttcttggca gtaggaagtg
20461 ttgaagggct ccatttcagg gagctttcca gagccgtctc agtagaggag ttagagagac
20521 ccggagcttg gtaaagtttc cgaagggttt ccctccctgt gcccttgctc tggtggccat
20581 gtgctctttc agaaggttct atcttggagt tcccattgtg gctcagcaga aacgagtctg
20641 actagcattc aagaggacac agcttcgatc cctggccttg ttcaatgggg ttaaagatcc
20701 agcattgccc tgagctgtgg tgtaggtcgc agctgtagct ctgattggat ccctagcctg
20761 ggaacctcca tatgtcacag atgcagccct aaaaagacga aaaaaaaaaa agaaaagaaa
20821 agaaacaaac aaacaaacac aagaagaaga aggcggctct gtcttcttcg aagtcagcaa
20881 ggccagggag agcaggacac tacagcttgt gtgggataca gctctccaga aagaatagaa
20941 tatttattta tttcgcccct gccttattct agaaaggaat tcagtccagg agagaaggtg
21001 tgagagggaa gaggcttgag gtcccagggc aggggccagg cagagggctc cggggaaatg
21061 atggcccagg gaggccaccc agcttggagg atgatcggcc tttgggcagg gatgtggagg
21121 cccagggtct gatccaaaag gtgctcagtg atactgacag aagacaagga cctgagtcca
21181 tgggttctca gaccagagga gttggtacag ctacacccag aagcccttga tggttcggct
21241 tcctggcggc ccagtgatca gcaacccatt ccagatggcc ggctgcaagt gtgacgggaa
21301 cacctttggc cgctactgca accactccaa ggacccctgt gatgagccgt gcttcccgaa
21361 tgtgaagtgc atttccggga agggctgcga ggcctgcccg acaaccctga ctggggatgg
21421 acgtcactgt gcacgtgagc tggggacagg gccttggcag gaggaggttc tggggatggg
21481 gcctaaacag ccaggaaaaa ccatttctcc cccctttcca gcaagtcttt agaatgttcg
21541 ataaaggggg ctatggtaag gaggtaaaca ggggagggga gggtgtgagg ccaaaggagg
21601 agagatggtc caggcatggg ggaggggcca gagtacagca gtcccaaggc aacgatgggg
21661 gaaaccaggg caaaagactg gagacaggga gtttccgtcg tggcgcagtg gttaacgaat
21721 ccgactagga accatgaggt tgcgggttcg atccctgtcc ttgctcagtg ggttaaggat
21781 ccagcgttgc cgtgagctgt ggtgtaggtt gcagacgcgg ctcggatcct gcgttgctgt
21841 ggctctggcg taggccagtg gctgcagctc cgattcgacc cctagcctgg gaacctccat
21901 atgccgcgag agcagcccaa agaaatagca aaaagaccaa aaaaaaaaaa aaaaaaaaaa
21961 aaagactgga gacaggaggg gacttccagc agcacccagg ggaccagagg taagctggct
22021 cctggaggtt tccagagatg caaggcccag ggcctctgca ttctccagtc aggacactgg
22081 cggcagggtg gggtgggggg gtctgccatg ggcatctagc agcaggaatg ccttgtcttc
22141 cctttgagtc ctccatccac catctgttcc ttgacctctg agccactgga gtcatgggtg
22201 gcatcatggt gacatcttca ctgagccccc taaaacactc aaatgaaatc tagaaaccac
22261 tgcatcttct gagcacagcc cagattggag aagcacagcc ttataaaggc caaaaagggg
22321 atggaagtga ggaaagcagg ctggtgggcc tttctcctcc cagccttttg ggtcctttcc
22381 tgatggggag ggtctaacaa cactccattc cctcagcaga agtgggagaa cccctacctg
22441 gcagaggaaa aagggttgtc tggcaaagaa atgtctggaa aagggagaaa gatggtttgg
22501 ggttcagagt ggcccacccg cagaacacag tttaagtgag ctttgcaggg aaaaagccaa
22561 ccctgtcctg ggagcagctt tctctggctg gcggctgcca gtgtggtgta gcagactgtc
22621 tgttccctct gcagctctca acagcttttt caactgttgg aacaagacct gtcctgagaa
22681 ttactgctac aaccagggtc gctgctacat ctcccagtct ctgacctgcg agcccgcctg
22741 cacctgcccc ccagccttca tggaagacac ccgctgcttc ctggctggaa aaaatttcac
22801 tccaaccatc cttccaggta cagccagatg ccctgccatc cccactccac acgtctctta
22861 acttggggga agatggggga ctctgggggc caggctcagt ccctcaaact cacaggacag
22921 aactccaaat ctctctcgtg tgctcgtaag ttgtaggctg tgggagtgag tggggggcag
22981 ctgtcacacc acaaaccaca ggtctgagca gtctgctgag aaaagactga aggggacctc
23041 acagctcagc gagacaagag tcaggccagc cttgatcaca gggtgaggtc agcagacctg
23101 gccatttctg attcttctgg tgaccttcta tgactgtaga agagggcagt cttcttgggc
23161 tgtcctcctt ggctaagtct gggtaaacct gtgaggtgac tatgaatctg cttcttccct
23221 ctccagagct tcccccaagg ctcatccagc tcttgctcag tgaaaaagaa aatgcctccc
23281 aagcagatgt caatgccacg gtcagtgctg caggctagct ctgggtgggg aggcggttct
23341 tggcagattc ttatccatgg cttgtacatt ctgcatctca tcatccatct agactcaact
23401 gccagaggtg gctagagcct cctgccccag gggggtgggg gtgagggagg cagggaggcg
23461 caggacagag acctccaaag ggctgtgaat gaccatctca caatgaacag aatctggggg
23521 cagggagagg ggggcaagtc aacacagcct cgcctctcct tctgagcatc cctcgagacc
23581 tgctcttggg cactatatgg aagaaacaca agagggcatt tcacctccag gaagccttcc
23641 ctgaattccc caagagaact gggggc tcct tcttcttggc tcccacaccc ctgttgattc
23701 tgcagtcaca ggcaaagtag cacaattgtt gagttactga ggctctagaa tcctacctgg
23761 gttcacatcc tggcagtacc actcactaga gcaggtatga tcattagaca aattatttga
23821 tctcccatat ataaaaacag aagtaataat agtactcatg tcaatgggtt gtgtcaaagt
23881 tgaaatgata atatatgtaa agcacttagc acagtgcctg aaacacaata cattcaaaaa
23941 ttaccaaaat aactattgtt tctgctgcta ctgctactac tactacaaca gcagcattct
24001 gggatctcta tggtaaaggc tgttgacatg gatctttgag tatgggcagc attctgggat
24061 ctctatggta aaggctgttg acatggatct ttgagtatgg gctgtttcta actcacactg
24121 acacacacac cccccccacc ccaacgcaca catctgacca tcccccaggt ctggctaaat
24181 ataattgatg ttcattcaga tatggggaga gggtgagtgg gtgagtcagg gatcagaggc
24241 aaggtagggg ctgttcattt catattctcg ctttgtgagt ggaactgggg tgaggttatg
24301 taaagtccta cactcaccaa caaacttcta ttaagtaggg acccatgcca gtcaccagtc
24361 gaggcttgtg gggcggggat gcagaaatag ataagaaaca ttctgagatt tcccatcatg
24421 gctcagtggt taacaagtcc gactaagaac catggggttg tgggtttgat ccctggcctt
24481 gctcagtggg ttaaggatcc ggcattgccg tgagctgtgg tgtaggttgc agacatggct
24541 cgggtcccac gttgctgtgg ctctagcgta ggccagtggc tacagctccc attcgacccc
24601 tagcctggga acctccatat gctgcgggag cagcccaaga aatggcaaaa agacgaacaa
24661 aaaaaaaatt ctggtggtcc tggggctcac attcaaagaa agaaactggg gtggaggggt
24721 gttcccattg tggctcagtg gtaacgaact gaactagtat ccataaggat gcgagtttga
24781 tccctggccc tcctcagtgg gttaaggatc tggcgttgcc gtgagctgta gtgtagactg
24841 acagctgcag ctccgattca acccctagcc tgggaacttt ctatatgcca tgggtgtggc
24901 cctaaaaaaa aagacaaaaa aaaaagaaga ataagaagga ggaggaggaa acacggatga
24961 atacatgcac aaaagtggga aaggtctcct ctctggactg gacctcaccc ccatttccac
25021 ccccacccca acccccaccc tctgccctag gtggcttata gactggggaa cctggatgtg
25081 cgggcctttc tccggaacag acaagtggaa cgagtgtaag tgggactgcc cccacctggg
25141 ccccggtcct tacctacacc cctcacccct cggcccctca ctgtgcccct ctcacacagt
25201 gaactcccag caataccggc ctcaggaaac ttccttcaat actggaaagt catctcggag
25261 ttccagtacc gccctggggg ccctgtcatt gacttcctga ataccaggct gctggacgca
25321 gtggtggagg cattcttatc ccaggcctca cagaggaggt ggaagagaag tgaggagccc
25381 aggaacaatg tggtcttcca acccatctcc aggagagatg tgctcagtgt gatggctggt
25441 gagtcactat ctcttggaca gcagcaagga tagctgaggg agcgggtaag cagagggagc
25501 ctgccacaga tctcaggccc acacaaaccc atggctgaag tgtattctac agcaggccac
25561 agacaagaag gaagtaaatt taggggggtt ttgtttgtca tttttaagag aaattgggaa
25621 ccttttaccc accagccttc tagattcagg aacaggagca caaagtactg cttgatgaat
25681 ccccacgtgt acagtttcac agagcaactg ctccaactgc tgctttcctg accattgctg
25741 ctaatggttt tctttctata aaattgttaa tgatttggtt aacatttatt ttataaatat
25801 aaacaaggac attgtttttg cctttgcctt cgcctccacc tggagatata aagttgaagg
25861 ctggagacct ggcccagggg cagaactatc acctccttga aagagcgtga ccctgtgcaa
25921 aagtccatcc ccagtcaagg cctcaacacg ctcatctgga aaaggaggtg ggatgtctga
25981 ggtccctttc agctctgaca gactctaatt atcccagaag gggaatgagg taggttggca
26041 aacttgccag cgaaggaaag gaatagccca ccctgtctta tgagtggggc agagaacagg
26101 acagtggcca tgccaaatgc cacagaaggt gctgtgttac ctgagcctag ggccaagcaa
26161 gagcttcttc tagaatccag gctaagctcc ctcccttcca ggggcaacaa tgagaaatcc
26221 tccaaactac tatggtatgt gtggtaggga ggcagtgttg gggataaggg aataaatact
26281 ccagacttga ctgtgccttt tcttagagaa aacaaatcaa caaaaccccc acaattttta
26341 ttatgcaagt acaatatgtt ttgtgtttag gacacagaag ttaacagaaa aaaaatcacc
26401 tgtattctaa tccccagaga caaatcccct ttgcacatac tctgctaaat tctcttttat
26461 gcataggaca cttcatatat caataaaggg atcaaatcat acctaatgtt tgataacctc
26521 tccattactt taactgtgaa catattacct tgtacattgt attcacattc tgatttgata
26581 tattatttta tgtcttaaat agtgtgtttt aatatatttt aaatagaatg tttttaaata
26641 tttttaaata ttttaaatag aatagtgact gttaaactga aggcttacta tgtcaggcac
26701 tgtgctaaat actgtgcata tattacctca tttaaacttt acagtaaggt agtttccttt
26761 taaaaataag gaaggaagtt cccgttgtgg ctcagcagta acaagcccaa ctagtatcca
26821 tgaggatgca ggttgaattc ctggcctcca tcagtgggtt aaggatctgg tgttgccatg
26881 agctgtggtg taggtcgcag acacgtttgg atcctgtgtt gctgtggctg tggctgtagc
26941 atagactgcc agctatagct cctattcaac ccctagccag ggaacttcca tatgccacag
27001 gtgtgaccct aaaaagcaaa ataaataaat aaaataaaag gaaatataaa ataaaaaata
27061 taaggaaact tataaaataa cgtaaaatat aaaataaaga aataaagaaa ctgaaggaat
27121 tctcttgtgg cacagcaggt gaaggatcca gtgtagtcac tgtagaggct taggttgctg
27181 ctgtggtgca ggttcaatcc ctggcctagg aacttccaca tgctataggc agaccaaaat
27241 ttaaacaaaa aaattttttt ttctttttgc catttgggcc gctccctcgg catatgtagg
27301 ttcccaggct aggggttgaa tcagagctgt aactgccggc ctacaccaga gccacagcaa
27361 tgcaggatcc gagccgtgtc tgcaatctac accacagctc acagcaatgc cgaatcctta
27421 acccactgag caaggccagg gatcgaaccc gaaaccccat cgttcccagt cggattcatt
27481 aaccactgag ccaccacagg aactccaata aagattttta ataaaaaaat aaggaaattg
27541 aggctgaaag agttaagaaa cttccccaaa ataacacaac tagttagtag tagagtttga
27601 atttcaactc aggtctagta gagttcaaag catatgctct taaccaatgc acttttcata
27661 ctgtggtcct catccattgc acaatattct ttatttaaca aaaccacttc tgggagctct
27721 cgttgtggct cagcgtgtta agaacctgac tagtatccaa aaggtttcgg ggtctatccc
27781 tggcctcgcc cagtgggttg gggatccagt attgcagcaa gctgtggtat aggttgcaga
27841 tgcagctcag atctggcatt gctgtggctg tggctgtggc tttggctggc agctgcagct
27901 ccaattcaac ccctagcctg ggaacttcca tatgccacta gtgtggccct aaaaaacaaa
27961 caaagccact tctgcttatt gttgctctta taaaccatgc tgctacagat ttccgtgaat
28021 atatgtagtt tttgtgtgtt tccaactatt tctcaggatg agtgcctaaa atggaattct
28081 tgacagaaaa ggtatagatt tggtgtaaat ccattggttt ccattttgct tgtgtcctat
28141 ctgacagtga atgtgagcac actgaaaaat tacttccaat gtaacggcta caaggactac
28201 aacctggttt tcagcccgga gagaggcttc acctgtgtgt ccccgtgcag caggggctac
28261 tgtgagaatg gaggccagtg ccagcaccta cccgaggggc cccaatgcag gtgagcaggg
28321 tgggggcagg aggtggaggg cagtgccaga gaatccaagc tgcaaggagt ctggaaatgc
28381 catggtacct cacagcgcct gaaaatgtag ggtggttccc tctgtaatcg agagaaaggg
28441 gatcagagaa aaagggacag aggaaaagaa gatatgagtg cctacatttg tggagaataa
28501 agatgctata taacctttgc acctcaaacc agcaaatgga tagatagata tgaccatttt
28561 aaaacgcaac ttagaaatga taaggaatct gactacggta cccagtaatt taagttacag
28621 gatagtgggt tccaaagcca aactcttcca cctgattttg cttctcgggt catggacagc
28681 ctactatctc cttgtggaaa aggaacccac tgtcctgtat ctaggccttg tcctggccca
28741 ccttaagatt cccagaaagg ggaggccctc tcaaactggt gccctctgtc cccttggttc
28801 atcaactggt ctgaggggct ccggtgagtg gagggcacta agttctgggc atgctaagga
28861 gaagcgcagt gcctcctggc gctcttgtca catcggtgcc ctccgtctac ctatggccct
28921 actacagtcg cagatgggct cagccctctc acctcccagg tctaagaagg cagcgggtgg
28981 agcacagcgt tgggggctgg gtgcagcttt ctgggagcta ggtgtgctca cctgcggcca
29041 actccgcctc cacctccacc ccccatccat gccctgctgt aggggtggag ctgaccccaa
29101 gtgaattcac taccagccca cctaggttgc tcagtggcct cggtttagca ccgcttgctg
29161 cctgcctgcc tgcaggggga ctgaccccat cccttcccca gagagggacc cagcctgttg
29221 tagaaaaatc tcctctggct gggctgagag caggggtggg gaggctaagg gccagcctcc
29281 ccatccccac accccgctgc tgaccggtcc tctctctctc tgtctccagc tgtgtgccct
29341 tttcatccct acgccctggg agcagcgct
2.MUC4分子标记在选育改良种猪抗腹泻病性状中的应用。
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603089B (zh) * | 2009-07-20 | 2011-08-10 | 江西农业大学 | 鉴别中国地方猪抗F4ac仔猪腹泻的分子标记及选育应用 |
| CN103525822A (zh) * | 2013-10-23 | 2014-01-22 | 湖南农业大学 | 一种仔猪腹泻抗性相关的分子标记检测方法及应用 |
| CN104878113A (zh) * | 2015-06-12 | 2015-09-02 | 江西农业大学 | 影响断奶仔猪的产肠毒素大肠杆菌f41性腹泻病抗性的主效snp标记及应用 |
| WO2018218857A1 (zh) * | 2017-06-02 | 2018-12-06 | 江西农业大学 | 改善猪肉质的myh4基因分子标记及在猪遗传改良中的应用 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003281988B2 (en) * | 2002-11-25 | 2008-06-12 | University Of Copenhagen | Porcine polymorphisms and methods for detecting them |
-
2008
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603089B (zh) * | 2009-07-20 | 2011-08-10 | 江西农业大学 | 鉴别中国地方猪抗F4ac仔猪腹泻的分子标记及选育应用 |
| CN103525822A (zh) * | 2013-10-23 | 2014-01-22 | 湖南农业大学 | 一种仔猪腹泻抗性相关的分子标记检测方法及应用 |
| CN103525822B (zh) * | 2013-10-23 | 2016-04-20 | 湖南农业大学 | 一种仔猪腹泻抗性相关的分子标记检测方法及应用 |
| CN104878113A (zh) * | 2015-06-12 | 2015-09-02 | 江西农业大学 | 影响断奶仔猪的产肠毒素大肠杆菌f41性腹泻病抗性的主效snp标记及应用 |
| CN104878113B (zh) * | 2015-06-12 | 2018-04-27 | 江西农业大学 | 影响断奶仔猪的产肠毒素大肠杆菌f41性腹泻病抗性的主效snp标记及应用 |
| WO2018218857A1 (zh) * | 2017-06-02 | 2018-12-06 | 江西农业大学 | 改善猪肉质的myh4基因分子标记及在猪遗传改良中的应用 |
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| Publication number | Publication date |
|---|---|
| CN101372712B (zh) | 2010-07-07 |
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