Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 360-362, 2013.
Article
in Zh
| WPRIM
| ID: wpr-318020
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>We expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.</p>
Full text:
1
Index:
WPRIM
Main subject:
Solubility
/
Virology
/
Gene Expression
/
Cell Line
/
Chemistry
/
Blotting, Western
/
Hemagglutinin Glycoproteins, Influenza Virus
/
Drosophila
/
Influenza, Human
/
Influenza A Virus, H1N1 Subtype
Limits:
Animals
/
Female
/
Humans
Language:
Zh
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2013
Type:
Article