Recombinant expression of human influenza A virus nucleocapsid protein and its antigenicity analyses / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 208-210, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-254102
ABSTRACT
<p><b>OBJECTIVE</b>To prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.</p><p><b>METHODS</b>NP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.</p><p><b>RESULTS</b>Three recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (12000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1640 to 11280.</p><p><b>CONCLUSION</b>The recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Vírus da Influenza A
/
Virologia
/
Sangue
/
Proteínas Recombinantes
/
Expressão Gênica
/
Proteínas do Nucleocapsídeo
/
Alergia e Imunologia
/
Escherichia coli
/
Influenza Humana
/
Genética
Tipo de estudo:
Estudo de rastreamento
Limite:
Animais
/
Humanos
/
Masculino
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2008
Tipo de documento:
Artigo
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