动物医学进展2017，Vol.38Issue(4)：7-12,6.
H9N2亚型禽流感病毒M2e蛋白在原核系统中的表达
Prokaryotic Expression of M2e Protein of H9N2 Subtype Avian Influenza Virus
摘要
Abstract
The plasmid containing the full length M2 gene of H9N2 subtype AIV was used as template for M2e gene amplification by PCR method;the various numbers of copies of M2e gene were constructed using the isocaudamer enzyme ligation of BglⅡand BamHI,and the various numbers of copies of M2e gene were connected into the vector pGEX-6p-1 forming five recombinant plasmid 2M2e-pGEX,4M2e-pGEX,6M2e-pGEX,8M2e-pGEX and 10M2e-pGEX;five recombinant plasmids were transformed into expression host strain,respectively.The bacterial liquid containing correct recombinant plasmids were induced by different IPTG concentrations and the sizes of the proteins were identified using SDS-PAGE method;the analysis of soluble protein was carried;the immuneoreactivities of five proteins were analyzed using Western blot method.The result showed that the expression levels of 2M2e-pGEX,4M2e-pGEX,6M2e-pGEX,8M2e-pGEX and 10M2e-pGEX reached the highest yield respectively under 0.25,0.25,0.5,0.25 and 0.75 mmol/L final concentration of IPTG after being cultured for 4 h at 37℃;the sizes of the recombinant proteins of 2M2e-pGEX,4M2e-pGEX,6M2e-pGEX,8M2e-pGEX and 10M2e-pGEX were 31.7,37.2,42.7,48.2 and 53.9 ku,respectively;five proteins existed in a form of inclusion body;Western blot analysis showed that the 2M2e-pGEX,4M2e-pGEX,6M2e-pGEX,8M2e-pGEX and 10M2e-pGEX proteins had a good specific reaction with anti-mouse monoclonal antibody against GST.These results lay a foundation for the further obtaining high immunogenicity protein and screening the universal immunogen.关键词
H9N2亚型禽流感病毒/M2蛋白胞外区(M2e)蛋白/重组蛋白Key words
H9N2 subtype avian influenza virus/extracellular domain of M2 (M2e) protein/recombinant protein分类
农业科技引用本文复制引用
张民秀,谢芝勋,黄莉,谢志勤,刘加波,谢丽基,邓显文,罗思思,黄娇玲..H9N2亚型禽流感病毒M2e蛋白在原核系统中的表达[J].动物医学进展,2017,38(4):7-12,6.基金项目
广西科技合作与交流计划项目(14123001-8) （14123001-8）
广西特聘专家专项经费项目(2011B020) （2011B020）
广西科技攻关重大专项项目(1222003-2-4) （1222003-2-4）
