Prokaryotic expression of a truncated form of bovine herpesvirus 1 glycoprotein E (gE) and its use in an ELISA for gE antibodies

Pesquisa Veterinária Brasileira

Stephan A.m. Oliveira²  Mário Celso S. Brum¹  Deniz Anziliero²  Odir Dellagostin¹  Rudi Weiblen²  Eduardo F. Flores² 

[1] ,Universidade Federal de Santa Maria Centro de Ciências Rural Departamento de Medicina Veterinária PreventivaSanta Maria RS ,Brazil

关键词: BoHV-5;    bovine herpesvirus;    vaccine;    DIVA;    recombinant protein;    BoHV-5;    herpesvírus bovino;    vacina diferencial;    proteína recombinante;   

DOI  :  10.1590/S0100-736X2013000100008

来源: SciELO

【 摘 要 】

This article describes the expression of a truncated form of bovine herpesvirus 1 (BoHV-1) glycoprotein E (gE) for use as immunodiagnostic reagent. A 651 nucleotide fragment corresponding to the amino-terminal third (217 amino acids) of BoHV-1 gE - that shares a high identity with the homologous BoHV-5 counterpart - was cloned as a 6×His-tag fusion protein in an Escherichia coli expression vector. A soluble protein of approximately 25 kDa purified from lysates of transformed E. coli was recognized in Western blot (WB) by anti-6xHis-tag and anti-BoHV-1 gE monoclonal antibodies. In addition, the recombinant protein was specifically recognized in WB by antibodies present in the sera of cattle seropositive to BoHV-1 and BoHV-5. An indirect ELISA using the expressed protein as coating antigen performed comparably to a commercial anti-gE ELISA and was able to differentiate serologically calves vaccinated with a gE-deleted BoHV-5 strain from calves infected with BoHV-1. Thus, the truncated gE may be useful for serological tests designed to differentiate BoHV-1/BoHV-5 infected animals from those vaccinated with gE-negative marker vaccines.

【 授权许可】

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