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.