Construction of eukaryotic expressing vector from ORF5/6/7 of porcine reproductive and respiratory syndrome virus.

Specific primers aiming to ORF5-ORF7 of porcine reproductive and respiratory syndrome(PRRSV) were designed.The target segments with restricted enzyme digestive sites were obtained by RT-PCR with PRRSV GD-XH and GD08-1 as template.All the segments were inserted into vector pEGFP-N1 to obtain six recombinant plasmids:pEGFP-N1-GD-XH-GP5,pEGFP-N1-GD-XH-M,pEGFP-N1-GD-XH-N,pEGFP-N1-GD08-1-GP5,pEGFP-N1-GD08-1-M,pEGFP-N1-GD08-1-N.The recombinant plasmid was transfected into Marc-145 cell.Fluorescence microscopy showed that EGFP were expressed in all transfected cells.