18型人乳头瘤病毒E6蛋白截短体（HPV18E6＊）的基因克隆与表达  

Cloning and Expression of Truncated Human Papillomavirus Type 18 E6 Protein （HPV18 E6＊）

作　　者：李小灵[1] 范立强[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《食品与药品》2013年第1期6-9,共4页Food and Drug

摘　　要：目的构建HPV18 E6*原核重组表达质粒,并优化其蛋白表达条件。方法以人宫颈癌HeLa细胞cDNA为模版,PCR扩增HPV18 E6*基因,构建重组表达载体HPV18 E6*-pET28a,在大肠杆菌BL21(DE3)中诱导表达重组蛋白,并优化其表达条件,SDS-PAGE检测重组蛋白表达状况。结果成功构建了重组表达载体;37℃表达,HPV18 E6*以不溶包涵体为主,表达量占菌体总蛋白的20%左右;15℃主要为可溶形式;包涵体变复性获得纯蛋白。结论 HPV18 E6*蛋白的高效表达为研究其蛋白作用机制及为研究预防和治疗子宫癌奠定基础。Objective To construct HPV18 E6＊ prokaryotic expression plasmid and optimize its expression. Methods HPV18 E6＊ gene was amplified by PCR with HeLa cell cDNA as template. After cloning it into expression vector pET-28a （＋）, HPV18 E6＊ protein was expressed by induction with IPTG in E. coli BL21 （DE3） and detected by SDS-PAGE. Results Recombinant expression vector was constructed and SDS-PAGE showed that recombinant HPV 18 E6＊ was about 20% of the total bacterial proteins, and was mainly expressed as inclusion bodies at 37 ℃ and soluble at 15 ℃. So the pure protein was obtained from inclusion bodies. Conclusion The highly expressed HPV18E6＊ protein provides fundamental basis for the further study on HPV18 E6＊ mechanism as well as prevention and treatment of uterine cancer.

关 键 词：人乳头瘤病毒 表达 大肠杆菌 

分 类 号：Q78[生物学—分子生物学]