遗传 ›› 2008, Vol. 30 ›› Issue (1): 59-64.

• 研究报告 • 上一篇    下一篇

家养水牛30个微卫星标记的多重PCR体系建立及其多态性检测

张毅, 孙东晓, 俞英, 张沅   

  1. 中国农业大学动物科技学院, 北京 100094

  • 收稿日期:2007-07-31 修回日期:2007-09-06 出版日期:2008-01-10 发布日期:2008-01-10
  • 通讯作者: 张毅

Optimized multiplex PCR sets and genetic polymorphism of 30 microsatellite loci in domestic buffalo

ZHANG Yi, SUN Dong-Xiao, YU Ying, ZHANG Yuan   

  1. College of Animal Science and Technology, China Agricultural University, Beijing 100094, China
  • Received:2007-07-31 Revised:2007-09-06 Online:2008-01-10 Published:2008-01-10
  • Contact: ZHANG Yi

摘要:

利用变性聚丙烯酰胺凝胶电泳结合硝酸银染色方法, 从30个微卫星标记中筛选出9组多重PCR体系, 其中4组三座位组合, 5组二座位组合; 在此基础上进一步检测了这些标记在中国海南兴隆水牛中的遗传变异性。结果显示其中26个标记具有多态性, 而其余4个标记(CSSM045, ILSTS008, RM099和HMH1R)为单态, 群体平均等位基因数4.7, 平均期望杂合度为0.534。所筛选出的多重PCR组合为家养水牛的群体遗传多样性 检测和亲子鉴定等研究提供了技术基础。

关键词: 多重PCR, 水牛, 微卫星

Abstract:

Nine multiplex-PCR-sets, including four triplex and five duplex combinations, were developed from 30 domestic buffalo microsatellite markers based on electrophoresis analysis of PCR products in denaturing polyacrylamide gels and visualization by silver-staining. Further, genetic variability on these microsatellites was examined in Xinglong buffalo of Hainan Province. Twenty-six loci exhibited genetic variation while four microsatellites (CSSM045, ILSTS008, RM099 and HMH1R) were monomorphism. The mean of number of alleles and the expected heterozygosity across 30 microsatellite loci were 4.7 and 0.534, respectively. These optimized multiplex-PCR-sets provide a technical basis for genetic diversity study and parentage test of domestic buffalo.