title: [Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification].
abstract: OBJECTIVE
  To express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.
  
  
  METHODS
  HA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.
  
  
  RESULTS
  We successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.
  
  
  CONCLUSIONS
  We expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.
authors.forename: Si-Min
authors.initials: SM
authors.lastname: Yao
authors.forename: Qiao
authors.initials: Q
authors.lastname: Lin
authors.forename: Guo-Liang
authors.initials: GL
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Deng
authors.forename: Guang
authors.initials: G
authors.lastname: Nie
authors.forename: Xue-Bao
authors.initials: XB
authors.lastname: Zheng
authors.forename: Ying-Xia
authors.initials: YX
authors.lastname: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004330:Drosophila; D005260:Female; D015870:Gene Expression; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000378:metabolism*; D007251:Influenza, Human / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D012995:Solubility
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus