日本脑炎病毒WHe株NS1基因的克隆、测序及表达  

Cloning, sequencing and expression of nonstructurai protein NSl of Japanese encephalitis virus strain WHe

作　　者：乔宪凤[1] 唐青海[2] 郑新民[1] 熊忠良[1] 刘西梅[1] 华文君[1] 周荆荣[1] 何维明[2] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所湖北省动物胚胎工程及分子育种重点实验室,湖北武汉430064 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2007年第4期23-26,31,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：湖北省自然科学基金项目(2005ABA195)

摘　　要：以日本脑炎病毒(Japanese encephalitis virus,JEV)WHe株的基因组RNA为模板,采用RT-PCR技术,克隆了JEV WHe株的NS1基因,并对其进行了测序和序列分析。构建了pET28b-NS1表达载体,转化表达宿主大肠杆菌BL21(DE3),并对其进行了诱导表达,对表达产物进行检测。结果表明,NS1基因全长1 145 bp,其核酸序列与JEV P3株同源性为99．4％,与SA14和SA14-14-2等27个JEV毒株的核苷酸序列同源性为98％,表明NS1基因的保守性很高;pET28b-NS1表达产物的相对分子质量约为43 ku,大小与预期结果相符。NS1基因克隆表达成功。Genomic RNA was separated from JEV Wile strain, and used as template for cDNA synthesis of NS1 gene. Then NS1 gene （1 145 bp） was amplified by RT-PCR. The analysis of sequence showed the nucleotide sequence of NS1 had the 99.4% identities with that of JEV P3 strain, and 98% identities with the NS1 reported in other 27 strains. Then the NS1 was cloned into the expression vector pET 28b （＋） to construct a recombinant prokaryotic expression plasmid, pET 28b（＋） NSl,and the recombinant plasmid was then transfected into E. coliBL21（DE3） . SDS PAGE electrophoresis showed the relative molecular weight of the expressed protein was about 43 ku in accordance with the presupposition.

关 键 词：日本脑炎 WHe株 NS1基因 RT-PCR 

分 类 号：S852.659.6[农业科学—基础兽医学]