title: [Construction and expression of eukaryotic expression vector of NS1 protein of influenza A (H1N1)].
abstract: OBJECTIVE
  To insert the full-length NS1 gene of influenza A (H1N1) into an eukaryotic expression vector PXJ40-HA, and to evaluate the expression of NS1 gene in transfected 293T cells.
  
  
  METHODS
  The NS1 gene of influenza A (H1N1) was amplified by RT-PCR and cloned into pMD18-T vector to construct a plasmid, named pMD18-T-NS1.The pMD18-T-NS1 and the PXJ40-HA were both digested using the same restrict enzymes and ligated, yielding the recombinant eukaryotic expression vector PXJ40-HA-NS1. The expression of the NS1 gene in transfected 293T cells was tested by Western blot.
  
  
  RESULTS
  The recombinant eukaryotic expression vector PXJ40-HA-NS1 was successfully constructed. The NS1 protein was observed to be expressed in 293T cells.
  
  
  CONCLUSIONS
  The full-length NS1 gene is obtained and its recombinant eukaryotic expression plasmid is successfully constructed. This study is of help to further understanding the biological function of NS1 protein and the mechanism of diseases induced by influenza A virus.
authors.affiliation: Key Laboratory of Enteric Pathogenic Microbiology, Ministry of Health, Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Nanjing 210009, China. wenshua925@163.com
authors.forename: Wen-shuai
authors.initials: WS
authors.lastname: Zhang
authors.forename: Qian
authors.initials: Q
authors.lastname: Bian
authors.forename: Tian
authors.initials: T
authors.lastname: Wen
authors.forename: Ying
authors.initials: Y
authors.lastname: Chi
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yong-jun
authors.initials: YJ
authors.lastname: Jiao
mesh_terms: D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010957:Plasmids / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C085820:INS1 protein, influenza virus; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins