Inhibition of Viral Repl ication by the Antisense RNA Targeting the 5′Regionsof Immediate2early Gene of Pseudorabies Virus

A 0.5kb antisense RNA fragment was designed targeting the 5’ non-coding region (NCR), translation initiation site and potential transcriptional active region of the sole immediate early gene(IE180) of Pseudorabies virus(PRV). The antisense RNA fragment was amplified by PCR and inserted into the downstream of hCMV IE promoter/enhancer of the eukaryotic expression vector pCI-neo. The resulting antisense RNA expression plasmid(pCI-0.5) was transfected into PK-15 cells and the transfected cells were selected by G418.The expression of antisense RNA was confirmed by RT-PCR. In order to assess the antiviral potency of the cell lines expressing the antisense RNA, the antisense RNA cell lines and the control cell lines were infected with 0.1 pfu of PRV strain Ea per cell. The infected supernatant were collected at different time points post-infection(p.i.) and the TCID\-(50) were further determined. The data showed that the cell lines expressing the antisense RNA could markedly inhibit the replication of PRV. The vira ˎ̥,Verdana,Arial">titers in ˎ̥,Verdana,Arial; mso-font-kerning: 1.0pt; mso-bidi-font-family: 'Times New Roman'; mso-fareast-font-family: 宋体; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA">cells expressing the antisense RNA was 1562 fold less than control cells at 40 hp i.