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.
Artigo
em Chinês
| WPRIM
| ID: wpr-318020
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 11280 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>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Solubilidade
/
Virologia
/
Expressão Gênica
/
Linhagem Celular
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Química
/
Western Blotting
/
Glicoproteínas de Hemaglutininação de Vírus da Influenza
/
Drosophila
/
Influenza Humana
/
Vírus da Influenza A Subtipo H1N1
Limite:
Animais
/
Feminino
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2013
Tipo de documento:
Artigo
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