重组人乳头瘤病毒11型L1蛋白在昆虫细胞sf9中的表达条件优化研究  

Optimization of conditions for expressing recombinant HPV11 L1 protein in sf 9 insect cells

作　　者：刘君[1] 张夕燕[1] 孙丽媛[1] 于世荣[1] 丁军颖[1] 许雪梅[2] 张振龙[1] 

机构地区：[1]北京生物制品研究所中试一室,100024 [2]中国医学科学院基础医学研究所,北京100730

出　　处：《国际生物制品学杂志》2009年第5期231-233,共3页International Journal of Biologicals

基　　金：国家高技术研究发展计划（2007AA02Z475）

摘　　要：目的确定利用昆虫细胞一杆状病毒表达系统表达重组人乳头瘤病毒11型L1蛋白（HPV11 L1）的最适表达条件。方法通过间接免疫荧光法鉴定目的蛋白表达情况；用蛋白印迹法（Western blot）确定在不同感染复数（MOI）、不同表达时间条件下目的蛋白的最适表达条件。结果当sf9细胞密度为2．0×10^6／ml时，以MOI值为2．0的重组杆状病毒接种，悬浮表达96h收获为最适表达条件。结论确定了目的蛋白的最适表达条件，为进一步大量表达及纯化提供参考。Objective To determine the optimum conditions for expressing recombinant human papillomavims 11 （HPV11） L1 protein using baeulovirus-inseet cell expression system. Methods The expression of the recombinant protein was identified by indirect immunofluorescenee assay, and the optimum conditions for expressing the recombinant protein at different multiplicities of infection （MOI） and time were determined by Western blot. Results The optimum conditions for expressing the recombinant protein were that the recombinant baculovirus was inoculated at MOI of 2.0 when the density of sf 9 cell was 2.0 × 10^6/ml, and the protein was expressed through suspension culture within 96 h. Conclusions The optimum conditions for expressing the recombinant protein are determined, which is to provide reference for further expression and purification in large quantity.

关 键 词：人乳头瘤病毒11 荧光免疫测定 L1蛋白 昆虫细胞 表达 

分 类 号：R3[医药卫生—基础医学]