Construction and expression of vectors carrying chemokine receptors CXCR4/CCR5 in SKOV3 cells

AIM: To construct the eukaryotic expression vector of human CXCR4 and CCR5,transfect SKOV3 cells with them.METHODS: The cDNA of CXCR4 and CCR5 was amplified by RT-PCR.After purification,the gene was cloned into a vector pEGFP.The sequence of inserted CXCR4 and CCR5 gene fragment was identified by enzyme digestion of EcoR I/Sal I and sequencing,and then the recombinant plasmid was transfected into SKOV3 cells which did not express CXCR4 and CCR5 by lipofectamine-mediated gene transfection method.The SKOV3 cells transfected with CXCR4 and CCR5 were examined by FCM.RESULTS: Vectors pEGFP-CXCR4 and pEGFP-CCR5 were obtained.Cell clones which were screened by G418 were obtained.The results of FCM indicatedthat transfected SKOV3 cells expressed CXCR4 and CCR5.CONCLUSION: SKOV3 cells that can express CXCR4 and CCR5 protein stably have been established successfully,which facilitates the researches of epithelial ovarian.