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Expression of porcine parvovirus VP2 gene requires codon optimized E. coli cells

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Abstract

Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the capsid protein VP2 of PPV was amplified and inserted into the plasmid pET-32a (+), which was then used to transform Escherichia coli Rosetta, the capsid protein of PPV was fused to a polyhistidine tag, and the position of the affinity tag is in N-terminus. VP2 was expressed using different expression host bacteria, including E. coli BL21, and Rosetta, and different plasmid vectors, including pET-30a (+), pET-32a (+), and pGEX-6p-1. After selection, only the fusion protein inserted into pET-32a (+) was expressed well in E. coli Rosetta. The recombinant bacterium produced high quantities of the fusion protein VP2, about 8% in total. The expressed VP2 was antigenically similar to the native capsid protein according to a Western blot assay performed with polyclonal antibodies obtained from pigs vaccinated with PPV. A simple, easily commercialized procedure was used to purify this protein. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of PPV and in the vaccination against PPV.

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Acknowledgments

The authors thank Mr. Chaofan Zhang for his help and constructive suggestions. The study was supported in part by funding from the National High-tech R&D Program (863 Program-2007AA100606).

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Authors and Affiliations

  1. Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 15001, China

    Ting Qi & Shangjin Cui

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  1. Ting Qi
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  2. Shangjin Cui
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Correspondence to Shangjin Cui.

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Qi, T., Cui, S. Expression of porcine parvovirus VP2 gene requires codon optimized E. coli cells. Virus Genes 39, 217–222 (2009). https://doi.org/10.1007/s11262-009-0378-6

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