H5N1亚型禽流感病毒M2的修饰表达及其免疫原性分析

Expression of Modified H5N1 Avian Influenza Virus M2 Gene and the Recombinant M2 Protein Immunogenicity Assay

禽流感的发生不仅会造成禽类的大量死亡,而且也严重地威胁着人类健康,接种疫苗是控制禽流感发生的主要措施之一.M2基因的保守性使其成为基因工程亚单位疫苗的目标抗原.本研究将M2基因的跨膜区删除后,构建原核表达载体pET32a-△M 2,IPTG诱导后,收获的细菌总蛋白SDS-PAGE电泳结果表明修饰的M2基因在原核表达系统中得到高效表达,表达蛋白以可溶性形式存在,Western杂交和动物免疫结果表明重组蛋白具有抗原性和免疫原性.本研究结果为利用M2重组蛋白开发具有交叉保护作用的禽流感疫苗奠定了基础.

The present vaccination is one of main strategy for control of avian influenza virus that critically threatened hnman health. Recent investigations of subunit component vaccine focused on M2 gene because M2 gent possessed a highly structurally conserved property among influenza viral strain. In this study, the recombinants prokaryotic expression plasmid pET32a-△ M2 harboring the deletion of the transmembrane segment of M2 gene was constructed. SDS-PAGE electrophoresis showed that modified M2 genc was highly expressed as fusion protein in a soluble form after genetic modified E.coli BL21（DE3） bacteria were induced with IPTG. The fusion protein of M2 possessed antigenicity and immunogenicity properties by Western Blotting analysis of fusion protein with standard antibody against M2（HSN1） and mouse vaccination of purified fusion protein respectively. The resnlts laid a foundation of further stndy on the development of cross-protection avian influenza vaccine.