title: [Subcloning of M1 gene fragment of H5N1 influenza virus and its expression in Escherichia coli].
abstract: OBJECTIVE
  To 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.
  
  
  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 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.
  
  
  RESULTS
  The recombinant Ml protein was recognized by antiserum against H5N1 subtype influenza virus, elicit specific antibody in immunized animals.
  
  
  CONCLUSIONS
  These confirmed that we successfully constructed the Escherichia coli vector expressing human H5N1 influenza virus M1 protein.
authors: State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
authors: Ai-jun
authors: AJ
authors: Chen
authors: Jian-qiang
authors: JQ
authors: Guo
authors: Li-hong
authors: LH
authors: Yao
authors: Cong-sheng
authors: CS
authors: Cheng
authors: Xiao-yu
authors: XY
authors: Liu
authors: Jin-qi
authors: JQ
authors: Fu
authors: Peng-wei
authors: PW
authors: Xu
authors: Zhi-qing
authors: ZQ
authors: Zhang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007114:Immunization; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000276:immunology; D007251:Influenza, Human / Q000276:immunology; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: C488936:M1 protein, Influenza A virus; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins