Establishment of recombinant adenoviruses for expressing PRRSV GP2a/GP2b,GP3,GP4,GP5,and M proteins

GP2a/GP2b-3,GP3,GP4,GP5,and M protein gene fragments of porcine reproductive and respiratory syndrome virus FJ-1a strain were amplified by RT-PCR and cloned onto shuttle vector pDC316.After co-transfection of one of the five recombinant shuttle vectors and the helper plasmid into 293 cells,recombinant adenoviruses Ad-GP2a/GP2b-3,Ad-GP3,Ad-GP4,Ad-GP5 and Ad-M were obtained.The titers of the five adenoviurses were 107.7-109.0 TCID50·mL-1.Among them,the Ad-GP5 titer was the lowest,at 107.7 TCID50·mL-1.The genome stability was determined by PCR detection of heterologous gene fragments in recombinant adenoviruses at 3rd,6th and 9th passages.In addition,when the recombinant adenoviruses were infected with the 293 cells,mRNA transcription of heterologous genes were confirmed by RT-PCR test.The establishment of recombinant adenoviruses for expressing PRRSV envelope proteins would provide a basic for further study on PRRSV genetic vaccine and the functions of the proteins.