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

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).