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Expression and antigenic characterization of the epitope-G1 of the Bovine ephemeral fever virus glycoprotein in Pichia pastoris

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Virologica Sinica

Abstract

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

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Authors and Affiliations

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China

    Fu-ying Zheng, Guo-zhen Lin & Chang-qing Qiu

  2. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150001, China

    Kui-zhang Yuan & Jun-ying Song

Authors
  1. Fu-ying Zheng
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  2. Guo-zhen Lin
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  3. Chang-qing Qiu
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  4. Kui-zhang Yuan
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  5. Jun-ying Song
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Correspondence to Chang-qing Qiu.

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Foundation item: National Dairy Foundation of China (2002BA518A04)

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Zheng, Fy., Lin, Gz., Qiu, Cq. et al. Expression and antigenic characterization of the epitope-G1 of the Bovine ephemeral fever virus glycoprotein in Pichia pastoris . Virol. Sin. 22, 347–352 (2007). https://doi.org/10.1007/s12250-007-0031-2

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  • DOI: https://doi.org/10.1007/s12250-007-0031-2

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