奶牛ZFX,ZFY基因片段的克隆、测序及PCR对奶牛性别鉴定灵敏度研究

The Cloning Sequencing of the ZFX and ZFY Genes Fragments and Sensitivity Studies of Amplification of These Fragments for Sex Identification in Cow by PCR

奶牛胚胎的性别鉴定是多年来奶牛业研究和探索的问题。尤其在体外授精,胚胎分割冷冻,保存及胚胎移植一系列生物科学高技术发展和应用的今天,这一问题的解决显得更加重要和迫切。用分子杂交的方法也可对胚胎进行早期性别鉴定,但这一方法必须要有大量的样品DNA,而且所需时间长。

According to the ZFY and ZFX genes in human, a set of primer ZF1 and ZF2 was designed. ZFY and ZFX fragments were amplified from cow DNA by PCR. Then ZFX and ZFY fragments were cloned to the pUC118 and sequenced from PCR fragments. According to these sequences, the oligonucleotide primer ZF3 in specific ZFY and the ZF4 in specific ZFX were designed. One. set of ZFX-specific primers was formed by ZF2 and ZF4. Other set of ZFY-specific primers was formed by ZF2 and ZF3. Nevertheless, the ZF3 and ZF4 can be used for allele-specific oligonucteotide probes (ASO). We used the two set specific primers or ASO to identify the sex of cow and the sensitivity reached 8pg DNA levels (correspond to the DNA of one cell).