牛早期胚胎性别鉴定的分子生物学方法研究  

Study of Nested PCR System for Sex Identification of Preimplantation Bovine Embryo

作　　者：周木清[1] 刘辉[1] 李邦佑 杨利国[1] 周世同 胡修忠[1] 吴世清 金尔光 杨翔 童勤[1] 王春芳 韩艳云 张淑君[1] 

机构地区：[1]华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉430070 [2]武汉市畜牧兽医科学研究所,武汉430065 [3]武汉市开隆高新农业发展有限公司,武汉430345

出　　处：《华中农业大学学报》2007年第1期67-70,共4页Journal of Huazhong Agricultural University

基　　金：武汉市科技项目(20042002059;20022002061;20067003111-06);国家农业科技成果转化资金(04EFN214200206);湖北省科技项目(2006CA023)资助.

摘　　要：根据牛SRY基因序列设计合成了2对嵌套式引物作为公牛特异性引物,同时根据牛β珠蛋白基因(HBB)序列设计了1对引物作为内参照引物,建立了巢式PCR反应体系。对50头母牛和30头公牛DNA样品的检测表明,母牛只能扩增出558 bp的HBB基因片段,而公牛则可以扩增出558 bp的HBB基因片段和219 bp的SRY基因片段,其鉴定结果与实际性别一致。同时,以微量的卵母细胞和精子细胞为模板对该反应体系的灵敏性进行了检验,结果表明,该反应体系可对10个细胞水平进行准确鉴定。Two pairs of SRY gene primer were designed as the nested primers for sex identification and one pair of HBB gene primer as an internal standard in this study and established the system of nested polymerase chain reaction （nested PCR）. DNA samples of 50 female and 30 male were tested, the results showed that only a 558 bp product of HBB gene could be visible for female, but for male, the 558 bp and 219 bp band could be visible. These results were as expected for both female and male. At the same time, oocytes and spermatids were used as the samples to check the sensitivity of this nested PCR The results showed that this method can successfully check the templates of ten cells.

关 键 词：性别鉴定 巢式PCR 卵母细胞 精子细胞 牛 

分 类 号：S823.2[农业科学—畜牧学]