摘要
从克隆、测序所得奶牛ZFY、ZFX序列中设计1对高特异性引物(或探针),即分别特异于牛ZFY、ZFX的ZF3、ZF4,与克隆时使用的引物ZF2搭配成ZF2、ZF3和ZF2、ZF4。利用这2对引物和ZF1、ZF2对奶牛基因组DNA进行NestPCR来判断性别,结果达到用超微量(8pg)DNA,即可检出ZFY、ZFX序列而判定雌雄的灵敏度;用非同位素标记试剂盒(DIG-System)标记ZF3、ZF4作为探针,对ZF1、ZF2扩增产物点杂交来判断性别。
The sequences of ZFY and ZFX gene fragments obtained from cloning and sequencing in bovine were analyzed. It was found that there were 88.7% homologous region within the two fragments. The remains appeared to be different. According to these different sequences, the primers or probe (ASO) ZF 3 specific for ZFY and the ZF 4 specific for ZFX were designed. One set of ZFY specific primers formed by ZF 2 and ZF 3 and the another set of ZFX specific primers formed by ZF 2 and ZF 4 were used to identify the sex of bovine by nest PCR. The results showed that a 8 pg DNA level of the sensibility for sex identification achieved in bovine by the nest PCR.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1997年第3期234-237,共4页
Chinese Journal of Veterinary Science
基金
复旦大学国家基因工程重点实验室项目
国家民委"八五"重点项目