非放射性标记ZF_3，ZF_4特异性探针对奶牛DNA杂交判断奶牛性别  

Sex Identification of Cows with Hybridization of Dig Oligonucleotide 3'-End Labeled ZF_3 and ZF_4

作　　者：徐亚欧[1] 龚荣慈[1] 

机构地区：[1]西南民族学院畜牧兽医系

出　　处：《西南民族学院学报（自然科学版）》1994年第4期400-403,共4页Journal of Southwest Nationalities College(Natural Science Edition)

基　　金：复旦大学遗传工程国家重点实验室资助课题

摘　　要：在测定了奶牛ＺＦＸ和ＺＦＹ基因的部分序列基础上，设计出两条分别特异于ＺＦＸ及ＺＦＹ的探针（ＺＦ＿４）及（ＺＦ＿３），同时也是两条引物。采用非同位素方法（ＤＩＧＯｌｉｇｏｎｕｃｌｅｏｔｉｄｅ３＇ｅｎｄｌａｂｅｌｉｎｇｋｉｔ）标记两条探针，并对ＺＦＸ，ＺＦＹ两个克隆子进行杂交，对以雄性奶牛和雌性奶牛的杂色体基因组ＤＮＡ为模板以ＺＦ＿１和ＺＦ＿２￣［１］为引物的ＰＣＲ产物进行杂支，结果表明ＺＦ＿３特异于ＺＦＹ，ＺＦ＿４特异于ＺＦＸ基因。并以该两条探针对不同浓度梯度的ＤＮＡ模板的ＰＣＲ产物进行点杂交，结果表明ＺＦ＿３和ＺＦ＿４对ＰＣＲ的产物的杂交来判断奶牛性别其灵敏度可达到８ｐｇＤＮＡ量的水平。On the base of testing panrt of the sequence of cows' ZFY and ZFX genes,two specificoligonucleotide ZF_3 and ZF_4 were designed,which were different from oligonucleotide ZFX and ZFY.The oligenucleotide ZF_3 and ZF_4 were two primers as well as two probes labeled with dig oligenucletide 3'-end.The different Consistent samples were the fragments of PCR corresponding to the 8ng , 800pg,80pg and 8pg Genomic DNA. The samples were hybridized with the ZFX specific“probe”or ZFYspecific“probe”. The results of bovine DNA sex dctermination assay show that the scnsitivity reaches8pg DNA levels(corresponding to the DNA of one cell).

关 键 词：性别鉴定 非同位素 杂交 乳牛 探针DNA 

分 类 号：S823.91[农业科学—畜牧学]