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.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
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)
