水牛胚胎性别鉴定的PCR方法  

Buffalo embryo sex identification using PCR amplification

作　　者：付强[1] 张明[1] 郑海英[1] 秦文松[1] 陆阳清[1] 卢晟盛[1] 卢克焕[1] 

机构地区：[1]广西亚热带生物资源保护利用重点实验室

出　　处：《中国兽医学报》2008年第9期1096-1100,共5页Chinese Journal of Veterinary Science

基　　金：广西科学研究与技术开发资助项目(0626001-3-1);利用性别控制技术快速繁殖良种奶水牛的研究资助项目(2005ZD05)

摘　　要：根据荷斯坦牛SRY基因设计1对引物扩增得到水牛SRY基因的全长序列2005 bp,对该片段测序后,设计2对特异于水牛SRY基因核心区的引物用于早期胚胎性别鉴定,同时根据水牛G3PDH基因设计2对引物作为内对照共同扩增,建立了多重巢式PCR体系并进行优化,使用优化后的PCR体系对取样后的27枚IVF胚胎和24枚Y精子受精胚胎进行检测,并使用斑点杂交检测扩增准确率。结果表明,27枚IVF胚胎均能有效扩增,雄性、雌性比例分别为51.9%(14/27)、48.1%(13/27),斑点杂交表明扩增准确率为100%;24枚Y精子受精胚胎的雄性比例为83.3%(20/24)。该巢式多重PCR方法具有很高灵敏度和稳定性,能够在实际生产中应用。Buffalo SRY gene was amplified by PCR with primers based on the sequence of Holstein cattle SRY gene.This fragment was sequenced,two pairs of nested primers were designed for sex identification which targeting to amplify swamp buffalo SRY conserved region.Simultaneously,Buffalo G3PDH gene was co-amplified to serve as a internal control for male and female embryos.Thus the multiplex nested PCR system was established and optimized.27 IVF embryos and 24 Y-bearing sperm fertilized embryos were examined after bi...

关 键 词：SRY基因 胚胎 PCR 性别鉴定 

分 类 号：S823[农业科学—畜牧学]