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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.
Artículo en Chino | 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>
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Solubilidad / Virología / Expresión Génica / Línea Celular / Química / Western Blotting / Glicoproteínas Hemaglutininas del Virus de la Influenza / Drosophila / Gripe Humana / Subtipo H1N1 del Virus de la Influenza A Límite: Animales / Femenino / Humanos Idioma: Chino Revista: Chinese Journal of Experimental and Clinical Virology Año: 2013 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Solubilidad / Virología / Expresión Génica / Línea Celular / Química / Western Blotting / Glicoproteínas Hemaglutininas del Virus de la Influenza / Drosophila / Gripe Humana / Subtipo H1N1 del Virus de la Influenza A Límite: Animales / Femenino / Humanos Idioma: Chino Revista: Chinese Journal of Experimental and Clinical Virology Año: 2013 Tipo del documento: Artículo