人乳头瘤病毒16型晚期蛋白在昆虫细胞中的表达

Expression of Human Papillomavirus Type 16 Capsid Proteins in Baculovirus-insect Cell System

目的 构建HPV1 6型晚期蛋白重组杆状病毒 ,并使其在昆虫细胞中获得高效表达。方法 首先用线性化的杆状病毒DNA与重组杆状病毒转移质粒共转染sf9昆虫细胞进行同源重组 ,获得 2株重组杆状病毒。结果 经鉴定该重组病毒中有目的DNA存在且可表达晚期蛋白 ;电镜观察发现重组杆状病毒感染的sf9细胞核内有乳头瘤病毒VLPs。结论 HPV1 6型晚期蛋白在昆虫细胞中的成功表达为HPV1

Objective To construct the recombinant baculoviruses containing L1 or L1 Plus L2 genes and express high-efficiency capsid proteins in insect cells.Methods By co-transfection of linearized baculovirus DNA and recombinant baculovirus transfer plasmids containing L1 or L1+L2 respectively,two recombinant baculovirus strains were gotten.Results The L1 and L2 genes of capsid protein were amplified by PCR in recombinant baculovirus;The 57KD L1 protein and the 95 KD,110KD L2 proteins were detected by Coomassie-blue staining and Western-blot;ME studies showed that there were the HPV16 virion in sf9 cells.Conclusion These results would be the basis to develop the HPV16 prophylactic vaccines and diagnosis kit.