丙型肝炎病毒抗原表位基因在新的抗原呈递系统中的重组构建及高效表达  

Overexpression of an epitope of HCV displayed in a new foreign epitope presenting system

作　　者：彭梅[1] 陈元鼎[1] 刘名英[1] 戴长柏[1] 

机构地区：[1]中国医学科学院/中国协和医科大学医学生物学研究所,昆明650118

出　　处：《中华传染病杂志》2001年第4期219-223,T001,共6页Chinese Journal of Infectious Diseases

摘　　要：目的　将人工合成的丙型肝炎病毒 (HCV)E1区的抗原表位基因 ,构建到一种新的抗原表位呈递系统的高效表达载体pET RNA2中进行高效表达。方法　运用基因工程技术将人工合成的HCVE1区的抗原表位基因 ,构建到一种新的抗原表位呈递系统的高效表达载体 pET RNA2中 ,构成一个嵌合基因进行高效表达。在该载体适当的位置插入外源抗原表位可使其呈现一定的空间构象 ,使抗原表位位于重组蛋白的表面并有望改善其免疫原性。结果　分别在载体的 10 6、15 3、3 0 5位氨基酸处插入外源抗原表位基因 ,成功构建了重组质粒 pET RNA HCV/A1、pET RNA HCV/A2和pET RNA HCV/A3 ,并分别在大肠杆菌中进行高效表达。表达产物相对分子质量约为 4 5 0 0 0。初步研究结果表明 ,在特定条件下不需要异丙基 β D硫代半乳糖苷 (IPTG)诱导就可使之获得高效表达 ,表达的嵌合蛋白占菌体总蛋白的 4 0 %以上。结论　HCV抗原表位基因成功构建到一种新的抗原呈递系统中并获得高效表达 ,为深入研究HCV抗原表位的免疫学和生物学特性奠定了基础 ,并对今后设计诊断试剂及抗原表位特异性疫苗有一定意义。Objective An epitope in E1 region of hepatitis C virus (HCV) was constructed and highly expressed in a foreign epitope presenting vector based on an insect virus, flock house virus capsule protein encoding gene as a vector. Methods Using genetic engineering techniques, an epitope in E1 region of HCV was constructed in a foreign epitope presenting vector based on an insect virus, flock house virus capsule protein encoding gene as a vector and lastly constructed a chimeric gene. The chimeric gene can be put into an expressing plasmid, and expressed in either eukaryotic or prokaryotic system. The foreign epitope can be expressed in this system on the surface of the vector stereotic structure with its native conformation, and which can improve its immunogenecity. Results An epitope in E1 region of HCV was successfully cloned in three positions (aa106、aa153、aa305) of a foreign epitope presenting system based on an insect virus, flock house virus capsule protein encoding gene as a vector.The recombinant epitope in this system could be highly expressed more than 40% of total cell protein in E.coli.BL21 cell in somewhat strict conditions without IPTG inducing. Conclusions We have made this chimeric gene highly expressed in E.coli.BL21, by the way different from routine method. The results suggest that the expression method and this epitope presenting system can be used in studying immunological and biological properties of HCV and developing diagnostic reagents as well as epitope-based subunit vaccines for controlling HCV infection.

关 键 词：丙型肝炎病毒 抗原表位 抗原呈递系统 HCV 重组构建 基因表达 

分 类 号：R373.21[医药卫生—病原生物学]