Expression and Antiviral Effects of Sus Scrofa Mx1 and Bos taurus Mx1 on PRV

Abstract

Objective: To study the ability of Sus Mx1 and Bos Mx1 to inhibit the replication of PRV.Methods: Sus scrofa Mx1 gene and Bos taurus Mx1 gene were cloned from IBRS-2 and MDBK cells to pc DNA3.1 / myc-His(-) B vector respectively,and transfected into PK-15 cells with LipofectamineTM2000 to detect whether they can express successfully in cells by RT-PCR and Western blot respectively. And then the cytotoxicity to cells WST-1 and the ability of Sus Mx1 and Bos Mx1 to inhibit the replication of PRV were checked. The PK-15 cells were infected with 20TCID50,100TCID50 PRV after transfection 24 h. Subsequently,samples were collected after infection 24 h,48h,72 h,and total viral copies were tested by extracting RNAs of frozen and thawed samples repeatedly. Meantime,the cytopathic effect after infecting 100TCID50 PRV in 72 h was observed. Results: both recombinant eukaryotic plasmids were constructed and expressed successfully. They were able to reduce the amount of RNA of PRV significantly by real time PCR( P 0. 01). Conclusion: Sus scrofa Mx1 and Bos taurus Mx1 can inhibit the replication of PRV in PK-15.