Objective:To investigate the expression of the recombinant plasmid of regulator of HIV-1 virion protein expression(Rev) in the eukaryotic cells,and to evaluate the effect on human herpesvirus 8(HHV-8) lytic cycle replication by Rev protein in BCBL-1 cells. Methods:After identification with enzyme digestion and nucleotide sequences analysis,the recombinant plasmid pRev-Flag was transiently transfected into BCBL-1 cells and NIH/3T3 cells. The expression of pRev-Flag mRNA and protein in both cells was detected by RT-PCR and Western blot,respectively. Subsequently,RNA was obtained from BCBL-1 cells transfected with pRev-Flag. RT-PCR was carried out to evaluate the expression of ORF26 mRNA(encoding the minor capsid protein) of HHV-8. Results:Nucleotide sequences analysis indicated that the cloned Rev sequence was 100% homology with Rev gene previously registered in GenBank. The specific band was detected at the expected place by both RT-PCR and Western blot. In addition,the expression of ORF26 mRNA in BCBL-1 cells transfected with pRev-Flag was weaker than the control. Conclusion:Rev gene was correctly expressed in BCBL-1 cells and NIH/3T3 cells,and Rev gene might inhibit KSHV lytic cycle replication.