Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification

VernacularTitle:应用果蝇S2细胞高效表达甲型H1N1流感病毒可溶性HA蛋白及其活性分析
Keywords: Drosophila; Influenza A virus; Hemagglutinins
From: Chinese Journal of Experimental and Clinical Virology 2013;27(5):360-362
CountryChina
Language:Chinese
Abstract: Objective To express soluble HA of A/H1 N1 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 × 103 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 × 103 D.After immunization with HA,the Abs titers were 1∶1280 and 1∶5120 respectively on 10 d,30 d.Conclusion We expressed soluble HA with good bio-activity,which contributed to research on immune diagnosis,subunit vaccine,and monoclonal Abs for influenza.