The Construction and Expression of PRV PK/gG/GFP Recombinant Transfer Vector

A transfer plasmid, KGDF was constructed from the plasmids containing part PK and gG genes.A report gene expression cassette ,GFP containing a multicloning sites ,was inserted bluntly into pPKgGD to construct a recombinant transfer vector,KGDF. That KGDF was constructed correctly was proven by restriction digestion and PCR analysis of GFP. KGDF was used to co-transfect BHK-21 cell together with the genome or the virus of PRV HB, and the recombinant virus with green fluorescence was picked using fluorescent microscope.The recombinant virus was further identified and purified using PCR of the gene GFP and pickup from the fluorescent cells.