Establishment Of Porcine CD151Transgenic PK-15Cells And The Suscepbility To PRRSV Infection

Porcine reproductive and respiratory syndrome virus mainly affects pigs macrophages, cause serious immunosuppression. In the cell culture, PRRSV can infect pulmonary macrophage and the African green monkey kidney cell line,such as MARC-145.CD151is PRRSV3’UTR rna-binding protein, which plays an important role in process of PRRSV infection MARC-145cells, whether the CD151involves in the same process in PAMs has not been reported. CD151was cloned by RT-PCR from PAMs using specific primers. Comparing with the reference sequences, the homology of cloned CD151reached99.7%. CD151was inserted into eukaryotic expression vector PCDNA3.0, named pCDNA3.0-CD151, which was then transfected into PK15cell and screened CD151+PK-15cells by G418resistance. Confirm the CD151+PK-15cells by RT-PCR. Infect MARC-145cell、CD151-PK-15cell and CD151+PK-15cell with VR-2332type of PRRSV and observe CPE periodically. Consequences shows that when MARC145appeared obviously CPE, CD151-PK-15cell and CD151+PK-15have no pathological changes, however CD151+PK15cell positive for immuno fluorescence antibody assay and RT-PCR ORF5gene test were positive. These results show that transfecting CD151gene can make PK15cell susceptible.