利用杆状病毒表达载体系统表达重组乙型肝炎病毒核心蛋白  

Expression of Recombinant HBcAg in Baculovirus Expression Vector System

作　　者：高琳琳[1] 王晓燕[1] 张颖慧[1] 李岩[1] 邓菲[2] 张艳芳[2] 林菊生[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝病研究所,武汉430030 [2]中国科学院武汉分院病毒研究所,武汉430071

出　　处：《华中科技大学学报（医学版）》2007年第6期787-790,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30330680)

摘　　要：目的利用杆状病毒表达载体系统制备重组乙型肝炎病毒核心蛋白。方法构建含有HBV C基因的杆状病毒转移载体pFastBac Dual-HBV core,转化大肠埃希菌DH10Bac进行转座,提取重组Bacmid DNA转染昆虫细胞Sf9,获得含有HBV C基因的重组杆状病毒。用重组杆状病毒感染Sf9细胞进行乙型肝炎病毒核心蛋白表达,然后通过SDS-PAGE电泳和Western blot分析表达情况。结果成功构建了含HBV C基因的重组杆状病毒,而且在昆虫细胞Sf9中表达了乙型肝炎病毒核心蛋白。结论利用Bac to Bac杆状病毒表达系统,成功地表达了乙型肝炎病毒核心蛋白,为进一步研究奠定了基础。Objective To produce recombinant HBcAg in insect cells by Baculovirus expression vector system. Methods HBV core gene was amplified by PCR and cloned into pFastBac Dual vector to construct the recombinant plasmid pFastBac Dual-HBV core, using pMD18-T Simple Vector as a mediator. Then the recombinant plasmid pFastBae Dual-HBV core was transformed into DH10Bac competent cells for transposition, after which the recombinant Bacmid DNA was isolated and transfected into Sf9 insect cells by lipofectamine. Three days later, recombinant baculovi.ruses were harvested to infect Sf9 cells and the expression levels were detected by SDS-PAGE and Western blotting after infection. Results The recombinant baculovirus containing HBV core gene was constructed successfully and HBeAg was expressed in insect cells at a high level. Conclusion The recombinant HBcAg was successfully expressed in insect cells with Bac to Bac expression system, which lays the foundation for further research.

关 键 词：乙型肝炎病毒核心蛋白 杆状病毒表达载体系统 昆虫细胞 

分 类 号：R329.29[医药卫生—人体解剖和组织胚胎学]