| 摘要: |
| 以日本脑炎病毒（Japanese encephlitis 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。NS11基因克隆表达成功. |
| 关键词: 日本脑炎 WHe株 NS1基因 RT-PCR |
| DOI： |
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| 基金项目:湖北省自然科学基金项目（2005ABA195） |
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| Cloning,sequencing and expression of nonstructural protein NS1 of Japanese encephalitis virus strain WHe |
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| Abstract: |
| Genomic RNA was separated from JEV WHe 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(+)NS1,and the recombinant plasmid was then transfected into E.coli BL21(DE3).SDS PAGE electrophoresis showed the relative molecular weight of the expressed protein was about 43 ku in accordance with the presupposition. |
| Key words: Japanese encephalitis virus WHe strain NS gene RT-PCR |
