Construction of a gE/gI-deleted mutant strain of epidemic porcine pseudorabies virus and its biological characteristics
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    Abstract:

    Objective To obtain a candidate vaccine strain against epidemic porcine pseudorabies virus.Method A gE/gI-deleted transferring plasmid pMD-LA-RA and a recombinant plasmid carrying EGFP gene were constructed according to the sequence of epidemic porcine pseudorabies virus. The homologous recombination was operated between pMD-LA-EGFP-RA and PRV AH, and then the recombinant mutant virus PRV AH gE-/gI-/EGFP+ was selected using EGFP as screening marker. In order to obtain the gE/gI-deleted mutant strain PRV AH gE-/gI- without the EGFP gene, the second homologous recombination was carried out between pMD-LA-RA and PRV AH gE-/gI-/EGFP+. The proliferation ability, genetic stability and immunogenicity of PRV AH gE-/gI- were evaluated by its growth curve, continuous passage in susceptible cells and animal immunization.Result PRV AH gE-/gI- was obtained through two homologous recombinations with fluorescence observation, plaque purification and PCR. The deletion of gE, gI genes and EGFP gene marker in PRV AH gE-/gI- was identified by PCR, fluorescence observation and sequencing. PRV AH gE-/gI- had similar proliferation ability to the parental strain and had good genetic stability and immunogenicity.Conclusion A gE/gI-deleted mutant strain of epidemic porcine pseudorabies virus with good immunogenicity was constructed successfully, which provides a basis for the development of gene deleted vaccine targeting epidemic strains.

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PAN Hui, LI Yanhua, XIANG Keyu, TANG Dong, CHENG Zhenzhu, LUO Yongwen, JU Chunmei. Construction of a gE/gI-deleted mutant strain of epidemic porcine pseudorabies virus and its biological characteristics[J]. Journal of South China Agricultural University,2018,39(2):9-15

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History
  • Received:October 30,2017
  • Revised:
  • Adopted:
  • Online: January 24,2018
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