High yield expression of duck hepatitis A virus VP1 protein in Escherichia coli, and production and characterization of polyclonal antibody

文献类型: 外文期刊

第一作者: Li, Chuanfeng

作者: Li, Chuanfeng;Chen, Zongyan;Meng, Chunchun;Li, Lu;Liu, Guangqing

作者机构:

关键词: Duck hepatitis A virus;Codon optimization;Escherichia coli;VP1

期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )

ISSN: 0166-0934

年卷期: 2013 年 191 卷 1 期

页码:

收录情况: SCI

摘要: VP1 protein, the capsid protein of duck hepatitis A virus (DHAV), contains critical epitopes for inducing a protective immune response. Due to its low-level expression in Escherichia coli (E. coli), the function of this protein is poorly characterized. In this study, a codon-optimized VP1 gene was chemically synthesized in terms of the codon usage bias in E. coli and subcloned into pET32a (+) to increase its expression. The recombinant VP1 fusion protein was purified from inclusion body by Ni2+ affinity chromatography His-Bind Resin and used to raise the rabbit anti-DHAV-VP1 polyclonal antibody. The expression of the codon-optimized VP1 gene in E. coli was significantly increased when compared to the wild-type VP1 gene, having an at least 17-fold increase. Western blot analysis showed that the recombinant protein was recognized by the rabbit anti-DHAV polyclonal antibody. Western blot also demonstrated that the rabbit anti-DHAV-VP1 polyclonal antibody could recognize the purified VP1 fusion protein specifically, and in the indirect immunofluorescent assays (IFA), the antibody was able to probe the VP1 protein in DHAV-1 infected cells. In conclusion, codon optimization increased dramatically DHAV VP1 expression in E. cob and the His-tagged VP1 fusion protein showed good antigenicity and immunogenicity. (C) 2013 Elsevier B.V. All rights reserved.

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