利用昆虫—杆状病毒表达系统表达人乳头瘤病毒16型L1蛋白

Production of HPV 16 major capsid protein L1 with baculovirus expression system

目的 获得人乳头瘤病毒 16型 (Humanpapillomavirustype 16 ,HPV16 )L1蛋白。方法 以pFB1为载体构建HPV 16L1杆状病毒表达质粒 ,并感染昆虫细胞Sf 9结果 收集 2 7℃ ,培养 72h的感染重组病毒的Sf 9,提取细胞蛋白。经SDS PAGE蛋白电泳分析 ,发现有一相对分子质量为 5 6 0 0 0的蛋白表达 ,Westernblot证实所表达的蛋白为HPV16L1。结论 昆虫—杆状病毒表达系统可有效地表达HPV

Objective To obtain human papillomavirus type 16(HPV 16) major capsid protein L1 with baculovirus expression system.Methods Using pFb1 as a vector,a recombinant baculovirus expressing plasmid,which contains HPV16L1 gene sequence,was generated.The constructed virus was infected into an insect cell line Sf 9.Results After incubating at 27℃ for 72 hours,the infected cells were collected and total cellular poteins were extracted.SDS\|PAGE assay revealed a roughly 56 000 expressed protein and Western blot confirmed that the expressed protein arose from HPV16L1.Conclusion HPV16 later protein L1 could be efficiently expressed with baculovirus expression system,and the expressed L1 protein remains to have good immunoreactivity.This study may supply a basic work for preparing virus\|like particle and prophylactic HPV subunit vaccines.