Objective:To construct the recombinant lentivirus containing HIV-1 Rev gene and detect the effect of Rev protein expression on the lytic cycle replication and latent infection of KSHV. Methods:The fragment of Rev gene from expression vector pCI-neo-Rev was cloned into the lentivirus vector pHAGE-CMV-MCS-IZsGreen,then the recombinant plasmid pLenti-Rev,packaging vector psPAX2 and envelope vector pMD2.G were cotransfected into the 293T cells. Culture media were harvested and filtered through a 0.45 μm filter to obtain the recombinant lentivirus. The viral titer was checked by observing the expression of IZsGreen protein. After infected with the recombinant lentivirus,the mRNA transcription and protein expression of Rev in 293T and BCBL-1 cells were detected by reverse transcription-PCR and Western blot,respectively. Then,Rev-Flag,Rta and vIL-6 protein from BCBL-1 infected with the recombinant lentivirus and transfected with pCI-neo-Rev were detected by Western blot. Further more,mRNA transcription of Rta in BCBL-1 transfected with pCI-neo-Rev was detected quantitatively by real-time PCR. Results:The recombinant lentivirus vector carrying Rev was constructed successfully with the viral titer of 2×107 TU/ml. BCBL-1 cells were efficiently infected by the recombinant lentivirus and Rev protein was readily expressed in these cells. The Western blot and real-time PCR results showed that Rta and vIL-6 expression were significantly upregulated by low Rev expression and vice versa. Conclusion:The lentivirus packaging system is available for BCBL-1,and Rev protein could express effectively in BCBL-1 infected with plenti-Rev. Forthermore,the Rev protein may regulate the lytic cycle replication and latent infection of KSHV in a complicated pattern.