[Construction and expression of vectors carrying chemokine receptors CXCR4/ CCR5 in SKOV3 cells].
OBJECTIVE
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.


CONCLUSIONS
SKOV3 cells that can express CXCR4 and CCR5 protein stably have been established successfully, which facilitates the researches of epithelial ovarian.
Department of Obstetrics and Gynaecology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Ting-Ting
TT
Xie
Li
L
Wei
Hui
H
Song
Yue-Cheng
YC
Yu
D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D019713:Receptors, CCR5 / Q000235:genetics*; D019718:Receptors, CXCR4 / Q000235:genetics*; D014162:Transfection
C527276:CXCR4 protein, human; D019713:Receptors, CCR5; D019718:Receptors, CXCR4