牛SRY同源基因的PCR扩增和克隆

PCR AMPLIFICATION AND CLONING OF A GENEOF CATTLE (Bos taurus) HOMOLOGOUSTO HUMAN SRY

本文采用人SRY基因的一对引物,通过PCR扩增获得了雄性牛(Bos taurus)SRY同源基因片段。进一步证实牛存在与人SRY基因同源的相应基因。将PCR产物与载体pUC—Eco—T连接后,用以转化JM109菌,经过与人SRY基因探针菌落杂交,筛选获得了牛SRY同源基因克隆pBosY O.6后者的插入片段为相应于人SRY基因保守区在内的一段约609bp DNA。此外还比较分析了人和牛SRY同源基因片段限制酶图谱。

Using human SRY gene primers, the gene fragment of cattle homologous to human SRY was amplified by PCR In male cattle, a gene fragment of about 609 bp homologous to human SRY was identified. By ligating pUC-Eco-T with the PCR products and colonies hybridization with human SRY probe, the positive colony with cattle the fragment was identified, and termed pBosYO. 6. The colony contains a about 609 bp insert corresponding to human SRY conserved ORF. The restriction map of the recombinant was analysed and compared with that of human SRY gene.