Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification.
- Author:
Si-Min YAO
;
Qiao LIN
;
Guo-Liang ZHANG
;
Hui YANG
;
Xiao-Feng DENG
;
Guang NIE
;
Xue-Bao ZHENG
;
Ying-Xia LIU
- Publication Type:Journal Article
- MeSH: Animals; Blotting, Western; Cell Line; Drosophila; Female; Gene Expression; Hemagglutinin Glycoproteins, Influenza Virus; analysis; chemistry; genetics; metabolism; Humans; Influenza A Virus, H1N1 Subtype; genetics; metabolism; Influenza, Human; virology; Mice; Mice, Inbred BALB C; Solubility
- From: Chinese Journal of Experimental and Clinical Virology 2013;27(5):360-362
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.
METHODSHA 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.
RESULTSWe 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.
CONCLUSIONWe expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.