寡肽转运蛋白PepT2原核表达质粒的构建及其表达初探  

Plasmid Constuction and Preliminary Prokaryotic Expression of PepT2

作　　者：赵东欣[1] 朱其丛[1] 薛永亮[1] 卢奎[1,2] 

机构地区：[1]河南工业大学化学化工学院,郑州450001 [2]河南工程学院,郑州451191

出　　处：《生物技术通报》2011年第4期133-136,共4页Biotechnology Bulletin

基　　金：国家自然科学基金项目(20772023);河南省创新型科技人才队伍建设工程项目(104200510022);郑州市创新型科技人才队伍建设工程资助项目(10LJRC174)

摘　　要：寡肽转运蛋白(PepT2,peptide transporter,SLC15A2)是哺乳动物体内能够转运二肽、三肽的蛋白。研究表明,一些类肽的小分子药物也是PepT2的底物,但PepT2的结构与生物学功能尚待研究。建立稳定表达PepT2的表达体系是研究PepT2的重要环节。根据GenBank中人PepT2基因序列,借助Primer5.0设计了1对寡核苷酸引物,经PCR合成长达2 190bp的目的序列,通过重组构建pET30a(+)/PepT2表达质粒,测序分析确认目的基因中的3个碱基发生突变。初步研究了pET30a(+)/PepT2在大肠杆菌BL21(DE3)pLysS中的表达,为PepT2原核表达的进一步科研和实际应用奠定了基础。PepT2 can transport dipeptide,tripeptide and some small molecule peptide-like drugs,but its structure and biological function need to be investigated.It is important to constuct a stable prokaryotic expression system of PepT2 in investigation of PepT2.A pair of primers was designed according to the hPepT2 gene with Primer5.0 software.The gene sequence of hPepT2 was synthesied by PCR and cloned into the expression vector pET30a（＋）.The recombinant plasmid pET30a（＋）/PepT2 was identified and analyzed,and three bases mutations in PepT2 gene were found.The recombinant plasmid pET30a（＋）/PepT2 was transformed into the BL21（DE3）pLysS expression system for expression,which established the foundation for the further scientific research and the real application.

关 键 词：PepT2 原核表达 基因 

分 类 号：Q78[生物学—分子生物学]