HBV全基因核心蛋白变异株稳定表达载体的构建及其抗原表达  

CONSTRUCTION OF STABLE EXPRESSION VECTORS WITH HBV GENOME CAPSID PROTEIN MUTANTS AND THEIR ANTIGENIC EXPRESSION

作　　者：陈伟红[1] 刘志华[1] 王九平 何海棠[1] 骆抗先[1] 

机构地区：[1]第一军医大学南方医院

出　　处：《解放军医学杂志》2003年第4期324-325,共2页Medical Journal of Chinese People's Liberation Army

摘　　要：通过定点突变技术将HBV质粒p3.8Ⅱ构建成核心蛋白变异株V6 0和L97。经序列测定和生物学活性检测后 ,亚克隆入EB病毒稳定表达载体。重组载体分别作内切酶酶切鉴定和序列测定证实 ,用脂质体介导转染HepG2 细胞稳定传代 ,定量测定各株培养上清HBV抗原表达。结果发现 ,3株HBsAg含量S/N值接近 ,变异株HBeAg含量S/CO值低于野生株。上述The site directed mutangenesis was performed to introduce capsid protein point mutations V60 and L97 into HBV genome plasmid p3.8Ⅱ. After identification of DNA sequencing and biological activities, the constructed plasmids were subcloned respectively into EB virus based vector for stable expression. After being confirmed by restriction enzyme analysis and DNA sequencing, the recombinant vectors were transfected into HepG 2 cells via the liposome. HBV antigen expression in culture supernatants of transformed HepG2 cells was quantified by Abbott kits. The values of HBsAg S/N of three reeombinants were at the similar level, while the values of HBeAg S/CO in culture supernatant of mutants were much lower than that of EBO wild type. The construction of these mutant vectors might act as a bascial tool to be used in studying the relationship between capsid protein mutations and HBV pathogenesis.

关 键 词：HBV 全基因核心蛋白变异株 稳定表达载体 构建 EB病毒 

分 类 号：R362[医药卫生—病理学]