[Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification].
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
; 27(5): 360-2, 2013 Oct.
Article
em Zh
| MEDLINE
| ID: mdl-24645323
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 11280 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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Expressão Gênica
/
Glicoproteínas de Hemaglutininação de Vírus da Influenza
/
Influenza Humana
/
Vírus da Influenza A Subtipo H1N1
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Female
/
Humans
Idioma:
Zh
Revista:
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
Assunto da revista:
VIROLOGIA
Ano de publicação:
2013
Tipo de documento:
Article