日本脑炎病毒NS1基因在大肠杆菌中的高效表达  

High-level Expression of Japanese Encephalitis Virus NS1 Gene in E.coli

作　　者：金吉东[1] 仇华吉[1] 田志军[1] 周艳君[1] 张守发[2] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]延边大学农学院动物医学系,吉林龙井133400

出　　处：《兽医大学学报》2003年第3期255-257,共3页

摘　　要：提取日本脑炎病毒弱毒株 SA1 4 - 1 4 - 2的基因组 RNA,用 RT- PCR扩增其 NS1基因 c DNA,将其克隆到原核表达载体 p ET- 30 b(+)中 ,转化大肠杆菌 BL - 2 1 (DE3) ,经 IPTG诱导 ,NS1基因获得高效表达。 SDS- PAGE分析显示 ,表达的融合蛋白表观相对分子质量约为 4 6 0 0 0 ,重组 NS1蛋白占菌体总蛋白的 30 .8%。 Western blotting分析表明 。The cDNA of NS1 gene was amplified by RT PCR from Japanese encephalitis virus(JEV) genomic RNA,and then cloned into an expression vector pET 30b(+),resulting in a recombinant plasmid pET30b NS1.The recombinant plasmid was transformed into E.coli strain BL 21(DE3).A His fused recombinant protein with an apparent molecular weight of about 46 000 was obtained after induction by IPTG as indicated by SDS PAGE analysis,accounting for 30 8% of the total bacterial cellular lysates.The recombinant protein showed reactivity with both monoclonal antibody against His tag and specific antisera against JEV in western blotting.

关 键 词：日本脑炎病毒 NSl基因 大肠杆菌 表达 乙型脑炎 

分 类 号：S852.65[农业科学—基础兽医学] Q786[农业科学—兽医学]