ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (10): 1064-1068.doi:

• 兽医 • Previous Articles     Next Articles

Construction of a Recombinant Pseudorabies Virus Coexpressing Porcine Parvovirus VP2 Gene and a Marker Enhanced Green Fluorescence Protein Gene

LUO Yan;GUO Wan-zhu; XU Zhi-wen; LIU Yan-li;YIN Hua-ping; WANG Xin;WANG Xiao-yu;GOU Lin   

  1. 1.Animal Biological Technological Center，Sichuan Agricultural University,Ya’an 625014,China;2.Biochemistry Laboratory, Sichuan Agricultural University, Ya’an 625014,China;3.Resource and Enviromental Sciences Department, Sichuan Agricultaral Univevsity Dujiangyan Campus,Dujiang yan 611830,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-10-25 Published:2005-10-25

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Abstract: The PPV VP2 gene obtained by PCR was inserted into pPI-2.EGFP vector, and the expression vector pPI-2.EGFP.VP2 was constructed. The results of Dot-blot hybridization and restriction enzymes digestion suggested that the construction of pPI-2.EGFP.VP2 was successful. PRV SA215 DNA and recombinant plasmid pPI-2.EGFP.VP2 DNA were co-transfected into Vero cells with calcium phosphate, and several recombinant viruses were screened out with fluorescence microscopy assay and Dot-blot hybridization. The recombinant virus named SA215-B was analyzed with BamHⅠrestriction enzyme digestion, Southern blotting, SDS-PAGE and Western blotting, the result confirmed that PPV VP2 gene had been inserted into the PRV SA215 DNA and expressed about 93 ku EGFP fusion proteins in recombinant virus. The successful construction of PRV-PPV recombinant virus would lay a foundation on the development of an genetically engineered PRV bivalent marker vaccine.