Subcloning of M1 gene fragment of H5N1 influenza virus and its expression in Escherichia coli.
- 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
- Publication Type:Journal Article
- MeSH: Animals; Cloning, Molecular; Escherichia coli; genetics; metabolism; Gene Expression; Humans; Immunization; Influenza A Virus, H5N1 Subtype; genetics; immunology; Influenza, Human; immunology; Mice; Recombinant Proteins; genetics; immunology; Viral Matrix Proteins; genetics; immunology
- From:Chinese Journal of Experimental and Clinical Virology 2011;25(4):254-257
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo generate the Escherichia col 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.
METHODSM1 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 coil BL21 (DE3) strain. The expression of M1 was induced by isopropy-beta3-D-thiogalactopyranoside. 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.
RESULTSThe recombinant Ml 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.
