摘要
从猪睾丸中提取RNA后,采用逆转录聚合酶链式反应(RT-PCR)方法获得PHGPx cDNA。扩增的目的基因片段经琼脂糖凝胶电泳回收,与pMD18-T载体连接,转入E.col i DH5α中筛选鉴定重组子。将质粒经酶切鉴定后,把酶切下的目的基因cDNA进一步克隆到真核表达载体pGAPZαA中,经PCR及序列鉴定,证实插入载体pGAPZαA中的片段为含有目的基因的核苷酸序列。真核表达重组质粒pGAPZαA-PHGPx cDNA的构建成功,有利于猪PHGPx的真核批量表达及对其生物学功能的进一步研究。
PHGPx cDNA was obtained from pig testicular using RT-PCR method after extracting RNA. The amplified PHGPx gene fi＇agment was connected with pMD18-T vector and transformed to E. coli DH5α . After the modification and identification of the plasmid, the PHGPx gene eDNA was cloned into eukaryotic expression vector pGAPZα A. Through PCR and sequence analysis, the fi＇agment in the vector pGAPZα A was proved to be PHGPx gene. Eukaryotic expression recombinant plasmid pGAPZα A-PHGPx cDNA was successfully constructed. It laid the foundation for the further study of pig PHGPx.
出处
《广东畜牧兽医科技》
2011年第6期36-37,40,共3页
Guangdong Journal of Animal and Veterinary Science
