利用昆虫-杆状病毒表达系统制备HPV16E6蛋白  

Expression of HPV16E6 protein with insect-baculovirus expression system

作　　者：郑瑾[1] 任斐[1] 余翔[1] 来宝长[1] 王一理[1] 

机构地区：[1]西安交通大学生物医学信息工程教育部重点实验室/西安交通大学疫苗研发中心,陕西西安710061

出　　处：《西北大学学报（自然科学版）》2008年第5期775-778,共4页Journal of Northwest University（Natural Science Edition）

基　　金：教育部博士点基金资助项目(20070698097)

摘　　要：目的利用昆虫-杆状病毒表达系统高效表达HPV16E6蛋白。方法将HPV16E6基因克隆入供体质粒pFastBac-HTb,然后再重组入杆状病毒基因组;用带有目的基因的杆状病毒转染昆虫(Sf-9)细胞,27℃培养72 h,收集被转染细胞,SDS-PAGE,Western-blot分析鉴定表达蛋白。结果SDS-PAGE分析显示所表达蛋白分子量约为24KD,Western-blot结果表明表达产物为HPV16E6蛋白。结论利用昆虫杆状病毒表达系统获得HPV16E6蛋白的高效表达。Aim To produce HPV16E6 protein with insect-baculovirus expression system.Methods HPV16E6 gene was cloned into donor plasmid pFastBac-HTb,then recombinanted to baculovirus genome.Sf-9 cells were infected with recombinant baculovirus containing the target gene coding HPV16E6 protein fusion with 6×His tag.After being cultured for 72 hours at 27℃,the post infected cells were harvested.The expressed protein was revealed and confirmed by SDS-PAGE and Western blot.Results The expressed HPV16E6 protein with 24KD was revealed by SDS-PAGE,and was confirmed by Western blot.Conclusion HPV16E6 protein has been efficiently expressed with insect-baculovirus system.

关 键 词：昆虫杆状病毒表达系统 重组DAN技术 HPV16E6 

分 类 号：R373[医药卫生—病原生物学]