Subcloning of M1 gene fragment of H5N1 Influenza virus and its expression in Escherichia coli

VernacularTitle:人H5N1流感病毒M1基因的亚克隆及其在原核细胞的表达
Author: Ai-Jun CHEN ¹ ; Jian-Qiang GUO ; Li-Hong YAO ; Cong-Sheng CHENG ; Xiao-Yu LIU ; Jin-Qi FU ; Peng-Wei XU ; Zhi-Qing ZHANG
Author Information

1. 中国疾病预防控制中心
Keywords: Influenza A virus; Genes; Gene expression
From: Chinese Journal of Experimental and Clinical Virology 2011;25(4):254-257
CountryChina
Language:Chinese
Abstract: Objective To generate the Escherichia coli vector expressing human H5N1 influenza virus M1 protein. To provide useful tools for detection of human H5N1 influenza virus and study on biological function of M1 protein. Methods M1 gene fragment was amplified by PCR using the influenza virus gene segment 7 as template, and was subcloned into pQE80-L vector. The recombinant plasmid pQE80-L/M1 was transformed into Escherichia coli BL21 ( DE3 ) strain. The expression of M1 was induced by isopropy-β-Dthiogalactopyranoside. We purified the recombinant M1 protein with polyhistidine tag with Ni2+ affinity chromatography. Mouse were immunized with the purified M1 protein for preparing antibodies against M1. Results The recombinant M1 protein was recognized by antiserum against H5N1 subtype influenza virus, elicit specific antibody in immunized animals. ConclusionThese confirmed that we successfully constructed the Escherichia coli vector expressing human H5N1 influenza virus M1 protein.