重组人乳头瘤病毒11型L1蛋白在昆虫细胞sf 9中的表达条件优化研究

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2.0×106/ml时,以MOI值为2.0的重组杆状病毒接种,悬浮表达96 h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.

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

Objective To determine the optimum conditions for expressing recombinant human papillomavirus 11 (HPVI1) L1 protein using haculovirus-insect cell expression system. MethodsThe expression of the recombinant protein was identified by indirect immunofluorescence 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 baeulovirus was inoculated at MOI of 2.0 when the density of sf 9 cell was 2.0 × 106/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.