悬浮培养昆虫细胞表达重组HPV16 L1蛋白  

Expression of HPV16 L1 protein in insect cell suspension culture system

作　　者：韩聪[1] 王燕[1] 商庆龙[1] 魏兰兰[1] 陈思佳[1] 

机构地区：[1]哈尔滨医科大学微生物学教研室,150081

出　　处：《中华实验和临床病毒学杂志》2007年第4期352-354,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：黑龙江省青年基金(QC06C062);黑龙江省科技攻关项目(GB02C111);哈尔滨科技攻关项目(2004AA3CS176-1)

摘　　要：目的用悬浮培养昆虫细胞方式表达人乳头瘤病毒16型（HPV16）L1蛋白，为获得病毒样颗粒（VLPs）及进一步深入研究疫苗和诊断试剂盒打基础。方法优化昆虫细胞Si9的悬浮培养条件；优化扩增病毒和蛋白的条件；空斑试验测定病毒滴度；SDS—PAGE和Western Blot分析目的蛋白的表达情况；透射电镜观察细胞内HPV16 L1蛋白形成的VLPs。结果优化后，悬浮培养昆虫细胞初始接种密度为5×10^5 cell／ml，以MOI（Multiplicity of Infection）=10接种重组病毒rBacV／HPVl6L1，72～84h收获细胞沉淀为最佳。经透射电镜观察，在重组病毒感染的昆虫细胞内，存在HPV16L1蛋白形成的VLPs。结论优化了细胞悬浮培养、病毒扩增和蛋白表达的条件；电镜观察在重组病毒rBacV／HPV16L1感染的昆虫细胞中，HPV16L1蛋白可形成VLPs。Objective To express the L1 protein of human papillomavirus type 16 （HPV16） in insect cell suspension culture system. Methods Optimized the conditions of suspension culture, recombinant virus amplification and protein expression. Determined the virus tilter by plague analysis and detected the target protein by SDS-PAGE and Western Blot; The formation of VLPs by HPV16 L1 protein was observed with TEM .Results The St9 cells could grow better in suspension culture with seeding density of 5 × 10^5 cell/ml and the maximum expression quantity was obtained by infection of cells with rBacV/HPV16L1 （MO1 = 10） and harvesting after 72-84 h. HPV16L1 protein could assemble into VLPs in St9 cells observed with TEM. Conclusion The conditions of cell culture, virus amplification and protein expression were optimized. HPV16 L1 protein could assemble into VLPs in St9 cells, which would provide a foundation for further study of the vaccine and diagnosis kits.

关 键 词：乳头状瘤病毒 人 蛋白质L1 细胞 培养的 病毒样颗粒 

分 类 号：R346[医药卫生—基础医学]