奶牛精子SRY基因原位杂交检测方法的建立  

Establishment of an Effective Approach to Sperm Fluorescent in Situ Hybridization for Sry Gene in Dairy Cow

作　　者：陈荣[1,2] 沈波[2] 高庆华[1,2] 

机构地区：[1]塔里木大学动物科学学院,新疆阿拉尔843300 [2]新疆生产建设兵团塔里木畜牧科技重点实验室,新疆阿拉尔843300

出　　处：《塔里木大学学报》2012年第4期32-35,共4页Journal of Tarim University

基　　金：新疆生产建设兵团博士资金专项(2010JC20);塔里木大学校长基金创新群体项目(TDZKCX201201)

摘　　要：Sry基因是Y染色体性别决定基因,采用primer premier 6对SRY基因序列设计引物,以公牛基因组DNA为模板扩增,高效DNA地高辛标记和检测试剂盒制备探针,对X精子(Y精子纯度7%)、Y精子(Y精子纯度93%)和常规精子(Y精子纯度50%)DTT去凝集、蛋白酶K去除鱼精蛋白,杂交。结果表明:在荧光显微镜下可见标本背景清晰,精子头部成蓝色,边界清楚,X精子4.67%(n=600)有荧光信号,Y精子86.33%(n=600)有荧光信号,常规精子46.17%有荧光信号。实验证明了所选取Sry基因序列为Y精子携带,建立了精子原位杂交方法。SRY gene is the special gene to determine the sex of Dairy Cow. SRY gene primers were designed by primer premier 6 And PCR applicatied with the Bulls genomic DNA was used astemplate. DNA labeled with high Digoxin and detection kit were used to produce SRY gene probes. X sperm （7% Y - sperm）, Y （93% Y - sperm） and conventional sperm （50% Y -sperm purity） were agglutinated with DTF, the protamine of X, Y and conventional sperm was removed with protease K, and X, Y and conventional sperm were hybridized, respectively. Results showed that： FISH images observed under fluorescence microscope for X, Y and conventional sperm with significantbackground, blue sperm head and clear boundary. The 4.67% sperm cells were found with fluorescent signal in X -sperm （n = 600） , 86.33% in Y -sperm （n = 600） , and 46.17% in conventional sperm. The experiment proved that Sry gene sequences were only carried by Y - sperm, rather than X - sperm. The method was established for sperm fluorescent in situ hybridiza- tion in dairy cow.

关 键 词：精子 荧光原位杂交 SRY基因 奶牛 

分 类 号：S823[农业科学—畜牧学]