A组轮状病毒外壳蛋白VP4的原核表达  

Expression of human group A rotavirus outer capsid protein VP4 in Escherichia coli

作　　者：石长信[1] 王长安[1] 王湛 周伟[1] 洪涛[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《中华实验和临床病毒学杂志》1996年第2期114-117,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家863高科技生物技术领域资助项目

摘　　要：选用原核表达载体 pGEX2T,该载体表达谷胱苷肽-S 转移酶融合蛋白,将完整的 A 组轮状病毒外壳蛋白(VP4)基因(2359bp)插入 pGEX2T 中,在 SDS-PAGE 上没有明显的表达条带,而将部分 VP4基因[5′端保留1269个碱基(KUVP4 1269)和631个碱基(KUVP4 631)],插入 pGEX2T·在SDS-PAGE 中有明显的表达条带。用 SDS-PAGE 分离的表达产物免疫豚鼠,能产生中和性抗体,表明KUVP4(1269)具有良好的免疫原性,而 KUVP4(631)不具有免疫原性。Three recombinant plasmids were constructed in which the full-length(2359bp)or partial(5'1269bp, 631bp)gene of KUVP4 were inserted into expressing vector pGEX2T,pGEX2T vector is used in bacterial systems to express foreign polypeptides as fusions with glutathione S-transferase(GST).When the full gene of VP4 was inserted into pGEX2T,no VP4 fusion protein was detected in SDS-PAGE.The partial genes containing 5'631bp or 1269bp of VP4 were successfully expressed as fusion proteins which are insoluble.The fusion proteins were purified by SDS- PAGE and the purified protein was used to immunized guinea pigs.Immunization of guinea pigs with the VP4(1269) induced antibodies that neutralized Wa(G1)and RV5(G2)rotaviruses.It suggests that VP4(1269)expressed in Es- cherichia coli contained VP4 antigenicity,whereas the VP4(631)showed no antigenicity at all in experimental ani- mals.The results show that the suitable length of VP4 can be expressed in Escherichia coil as fusion protein with GST.The fusion protein still retain the antigenicity of VP4.

关 键 词：VP4 A组 轮状病毒 外壳蛋白 原核表达 抗原 病毒 

分 类 号：R373[医药卫生—病原生物学]