CAJ | 학술논문

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PHGPxcDNA。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．coli 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pGAPZ旺A中，经PCR及序列鉴定，证实插入载体pGAPZαh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-PHGPxcDNA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。
종저고환중제취RNA후，채용역전록취합매련식반응（RT—PCR）방법획득PHGPxcDNA。확증적목적기인편단경경지당응효전영회수，여pMD18-T재체련접，전입E．coli DH5α중사선감정중조자。장질립경매절감정후，파매절하적목적기인cDNA진일보극륭도진핵표체재체pGAPZ왕A중，경PCR급서렬감정，증실삽입재체pGAPZαh중적편단위함유목적기인적핵감산서렬。진핵표체중조질립pGAPZαA-PHGPxcDNA적구건성공，유리우저PHGPx적진핵비량표체급대기생물학공능적진일보연구。
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.