大熊猫SRY基因的PCR扩增和克隆

PCR Amplification and Cloning of Giant Panda(Ailuropoda melanoleuca) SRY Gene

本文采用人ＳＲＹ基因的一对引物，通过ＰＣＲ扩增获得了雄性大熊猫ＳＲＹ基因片段。表明大熊猫存在与人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大熊猫ＳＲＹ基因克隆，命名为ｐＡＭＹ０．６，其插入片段为相应于人ＳＲＹ基因保守区在内的一段约６０９ｂｐＤＮＡ。此外，还制作和比较分析了人和大熊猫基因片段的限制酶图谱。

Using human SRY gene primers, the gene fragment of giant panda homologous to human SRY was amplified by PCR. In male giant panda, a gene fragment of 609bp homologous to human SRY was identified. By ligating pUC Eco-T with the PCR products of the SRY gene of giant panda and colony hybridization with human SRY probe, the positive clony with giant panda SRY fragment was identified, and the chimera plasmid has been termed pAMY0.6. The plasmid contains a 609bp insert corresponding to human SRY conserved ORF. The restriction map of the recombinant was analysed and compared with that of human SRY gene.