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[利用PCR-RFLP技术对小梅山猪、中梅山猪和大白猪SLA-DQB基因外显子2的多态性分析]

[Polymorphism analysis of the exon 2 of SLA-DQB gene in Xiao Meishan, Zhong Meishan and Yorkshire pigs with PCR-RFLP].

作者信息

Fang Mei-Ying, Hu Xiao-Xiang, Li Ning, Wu Chang-Xin

机构信息

College of Animal Science and Technology, China Agricultural University, Beijing 100094, China.

出版信息

Yi Chuan Xue Bao. 2002;29(8):685-7.

Abstract

SLA is an important genetic region which influences disease resistance in pigs, and the gene family is one of important candidates for disease resistance and susceptibility. In present research the exon 2 of SLA-DQB gene was amplified and a uniform fragment of 273 bp was obtained in Xiao Meishan, Zhong Meishan and Yorkshire Pigs. The 273 bp PCR product was digested with restriction endonuclease Rsa I and genetic Polymorphism was investigated with PCR-RFLP (restriction fragment length polymorphism). Polymorphic sites were detected at base position 11, 94 and 124 of the exon 2 of SLA-DQB gene. There were two kinds of genotypes in Xiao Meishan pigs, 246 bp/27 bp(AA) and 132 bp/84 bp/30 bp/27 bp(BB), the frequencies of genotype AA and BB are 0.571, and 0.439 respectively. Four kinds of genotypes were found in Zhong Meishan pigs, 246 bp/27 bp (AA), 132 bp/84 bp/30 bp/27 bp(BB), 132 bp/114 bp/27 bp(CC) and 246 bp/132 bp/84 bp/30 bp/27 bp(AB), their frequencies are 0.037, 0.593, 0.037 and 0.333 respectively. Five kinds of genotypes were found in Yorkshire pigs, 246 bp/27 bp(AA), 246 bp/132 bp/114 bp/27 bp(AC), 132 bp/114 bp/27 bp(CC), 246 bp/132 bp/84 bp/30 bp/27 bp(AB) and 273 bp/246 bp/27 bp(AD), the frequencies of five genotypes are 0.467, 0.300, 0.100, 0.100 and 0.033 respectively. Statistical results indicated that there is no favorable allele in Xiao Meishan pig breed; B is the favorable allele in Zhong Meishan and its gene frequency is 0.759; while in Yorkshire pigs A is the favorable allele and its gene frequency is 0.683. Four alleles (A, B, C and D) were found in SLA-DQB locus among three pig breeds; The results of chi 2 showed that four alleles of this locus fit with Hardy-Weinberg equilibrium. The research showed that conservation of Chinese local pig breeds is necessary in order to protect our rich genetic resources and make use of them in the future.

摘要

SLA是影响猪抗病性的一个重要遗传区域,该基因家族是抗病性和易感性的重要候选基因之一。在本研究中,扩增了SLA - DQB基因的外显子2,在小梅山猪、中梅山猪和大白猪中均获得了一条273 bp的一致片段。用限制性内切酶Rsa I对273 bp的PCR产物进行酶切,并采用PCR - RFLP(限制性片段长度多态性)技术研究遗传多态性。在SLA - DQB基因外显子2的第11、94和124位碱基处检测到多态性位点。小梅山猪有两种基因型,246 bp/27 bp(AA)和132 bp/84 bp/30 bp/27 bp(BB),基因型AA和BB的频率分别为0.571和0.439。中梅山猪发现四种基因型,246 bp/27 bp (AA)、132 bp/84 bp/30 bp/27 bp(BB)、132 bp/114 bp/27 bp(CC)和246 bp/132 bp/84 bp/30 bp/27 bp(AB),它们的频率分别为0.037、0.593、0.037和0.333。大白猪发现五种基因型,246 bp/27 bp(AA)、246 bp/132 bp/114 bp/27 bp(AC)、132 bp/114 bp/27 bp(CC)、246 bp/132 bp/84 bp/30 bp/27 bp(AB)和273 bp/246 bp/27 bp(AD),五种基因型的频率分别为0.467、0.300、0.100、0.100和0.033。统计结果表明,小梅山猪品种中没有优势等位基因;B是中梅山猪的优势等位基因,其基因频率为0.759;而在大白猪中A是优势等位基因,其基因频率为0.683。在三个猪品种的SLA - DQB位点发现了四个等位基因(A、B、C和D);卡方检验结果表明该位点的四个等位基因符合哈迪 - 温伯格平衡。研究表明,有必要保护中国地方猪品种,以保护我们丰富的遗传资源并在未来加以利用。

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