1.Key Laboratory of National Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Science, Haerbin 150001，China; 2.Collge of Veterinary Medicine, Nanjing Agriculture University, Nanjing 210095，China
Abstract：Vaccine strain of Goatpox virus (GPV) cultivated and widely used in domestic was employed as the mother virus. The tk gene of GPV, the nonessential region during virus replication, was chosen as target site and homologous arm when inserting foreign gene. The stability of lacZ and gfp genes expression in recombinant rGPV-lacZ-gfp downstream of back to back promoter 11 and promoter 7.5 was testified after recombination of the transfer vector pTK-lacZ-gfp with the mother GPV, then lacZ gene in pTK-lacZ-gfp was replaced by gpt gene in order to obtain a purified recombinant GPV expressing foreign gene stably (rGPV-gpt-gfp) by using MPA blocking off the nucleic acid metabolize. Further more recombinant GPV construction system and optimized the purification methods were established, which supply a technique platform for investigation of live vector vaccine and relative basal research.
