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

摘要: 目的构建HPV16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 HPV16型晚期蛋白在昆虫细胞中的成功表达为HPV16型预防性基因工程亚单位疫苗的研制和诊断试剂的研究开发奠定了基础.

Abstract: 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 baculovir us 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 HPV 16 prophylactic vaccines and diagnosis kit.