Constrction Of Shuttle Vector Plasmid Expressing EGFP Of Pseudorabies Virus-Fa Strain

By means of restriction enzyme digestion, ligation and insertion the shuttle vector plasmid pPI-2.EGFP of Pseudorabies virus-Fa strain (PRV Fa) was obtained based on plasmid pPB7. PRV Fa strain BamHI-7 was digested with Nco\. deleted gE, 11k and part of gI and 28k genes, was ligated with T₄DNA Ligation. After translated and filtrated, we got PPB7-1: after digested with SalI. PPB7-1 deleted gG and part of gD. we got pPI: after digested with EcoRI and BamHI, deleted EcoRI, BamHI, HindIII enzyme sites. pPI turned to pPI-1; after digested with KpnI, pPI-2 was got; afterward EGFP and its integrality gene expression case which lie in the plasmid pEGFP-C1 were inserted into StuI site which lies in gI gene startup backward site of pPI-2. recombinants were filtrated in the culture medium, pPI-2.EGFP was got which contains four multi-cloning sites. Identified using the mothods of enzyme digestion and Dot-bolt, the result indicated the construction of pPI-2.EGFP was successful.