摘要
目的:构建H5N1流感病毒基质蛋白M2基因的原核表达系统,为研究其在体液免疫中的作用奠定物质基础。方法:应用PCR方法从流感病毒H5N1cDNA中扩增出M2的全基因序列,插入至表达质粒载体pGEX6P-3上,构建重组原核表达质粒pGEX-6p-3/M2。重组质粒经测序分析确认后,转化感受态大肠杆菌BL21,经IPTG诱导表达M2蛋白后,对其表达产物进行SDS-PAGE和Western Bloting分析。结果:PCR扩增的M2基因长度为297bp,经酶切和测序分析,插入到载体的基因与GeneBank公布的序列相似性达100%;SDS-PAGE和Westem Bloting分析显示,经IPTG诱导出分子质量为37.2 KD的目的蛋白。结论:成功构建了H5N1流感病毒基质蛋白M2原核表达载体pGEX-6p-3/M2,并在大肠杆菌中获得表达。
Objective:To clone and express the gene of matrix protein 2(M2) of H5N1 influenza virus,and to establish material foundation for the study of function mechanism in body fluid immunity.Methods:The M2 gene whose length was 297 bp was amplified by PCR from cDNA of influenza virus,and cloned into the expression vector pGEX-6P-3.Recombinant plasmid was then transformed into E.coli BL21,which was induced with IPTG.The whole protein of E.coli was analyzed by SDS-PAGE and western bloting.Results:M2 protein of influenza A virus,whose weight was 37.2 kD was successfully cloned and expressed in the E.coli.Conclusion:M2 protein acquired good fusion expression,which can provide basis for the further study on its immunity mechanism.
出处
《中国卫生检验杂志》
CAS
2011年第8期1920-1922,共3页
Chinese Journal of Health Laboratory Technology
基金
广州市医药卫生科技项目(2007-YB-157)
关键词
流感病毒
M2蛋白
克隆表达
Influenza virus
Matrix protein2
Clone and expression