Construction Of Recombinant Goatpox Virus Expressing Green Fluorescence Protein And Expression The G Protein Of Bovine Ephemeral Fever Virus In The Recombinant Vaccinia Virus

Goatpox virus (GPV) mainly infect goat, sheep, cattle and other ruminates but other mammals. When infected GPV, mankind usually wouldn' t show resious disease, even any symptom in infected person. Because GPV genome is charactered by high inheritance stability and strongly tolerance to foreign gene, it can contain a gene long up to 25kb but still retain infectivity. GPV plays an amazing role in the researching fields of virology, immunology and vaccinology. Recently several antigen genes from ruminate have been successfully expression in GPV vector. Recomabinant GPV displayed a prospect in ruminate disease control.Vaccine strain of Goatpox virus (GPV) cultivated and widely used in domestic animals was employed as the mother virus. Tk gene of GPV nonessential region during virus replication was chose as target site and homologous arm when inserting foreign gene. The stability of lacZ gene and eGFP gene expressed 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 we established recombinant GPV construction system and optimized the purification methods which supplied a technique platform for investigation of live vector vaccine and relative baSal research.Based on pSC11 plasmid, a transfer vector expressing Glycoprotein of recombinant viccinia virus(BEFV) was established. The vector clone into back to back promoter 11 and promoter 7.5 of pSC11 plasmid, lacZ LacZ gene and BEFV G gene were separately cloned into downstream of promoter 11 and promoter 7.5 of pSC11 plasmid. The recombinant viccinia virus transfered vector pSC11-BEFV-G was cotransfected with WR stain of VV at HeLa cell by calcium phosphate-DNA coprecipitation method. The sensitivity of the recombinant virus to specific serum was studied by indirect immunofluoresence assay (IFA). The result showed that recombinant BEFV G protein was confirmed to possess a specific reaction ability to anti-sera.