Construction and Identification of Recombinant Goatpox Virus Expressing CDV H Gene

CHENG Song1 WANG Hexing2 ZHENG Min3 LUO Manlin4 SHAN Fen1 LI Wanping1 CHEN Wu1

(1.Guangzhou Zoo, Guangzhou, China 510070)
(2.Mengzi Animal Disease Prevention and Control Center, Mengzi, China 661199)
(3.Guangxi Center for Animal Disease Control and Prevention, Nanning, China 530001)
(4.College of Veterinary Medicine, South China Agricultural University, Guangzhou, China 510642)
【Novelty】Through homologous recombination recombinant goatpox virus was generated after purification and polymerase chain reaction. The immunofluorescence test and western blotting demonstrated that the H gene of the Canine distemper virus(CDV) had been inserted into the goatpox virus and correctly expressed the H protein of the CDV in cells.The recombinant GTPV AV41 is named as vpTK-H-Eg.

【Abstract】In this study, the H gene sequence of the canine distemper virus (CDV) in giant panda was modified and synthesized based on the published coding sequences on GenBank. First, the gene transfer vector pMDTK-PEL was constructed. The H gene from the CDV was cloned into the transfer vector pMDTK-PEL through digestion with restriction enzymes, then the H gene expression cassettes were inserted into the vector pTK-Eg, thus we obtained the recombinant transfer vector pTK-H-Eg. The recombinant transfer vector was transfected into BHK cells that had been infected with the goatpox virus. The recombinant goatpox virus containing the H gene of CDV was cultured in Vero cells which then underwent pressure selection by adding mycophenolic acid to the culture. After purification and polymerase chain reaction, the immunofluorescence test and Western blotting demonstrated that the H gene of CDV had been inserted into the goatpox virus and correctly expressed the H protein in cells. We obtained the recombinant GTPV AV41 and name it vpTK-H-Eg.

【Keywords】 Canine distemper virus (CDV); Homologous recombination; Recombined goatpox virus;

【DOI】

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This Article

ISSN:1000-8721

CN: 11-1865/R

Vol 34, No. 04, Pages 550-556

June 2018

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Abstract

  • Materials and methods
  • Results
  • Discussion
  • References