Construction and identification of a luciferase reporter vector containing human CXCR4 promoter

Objective To construct and identify a luciferase reporter vector pGL3-neo-CXCR4 containing human CXCR4 promoter.Methods The gene Human promoter of CXCR4 was obtained by PCR amplification and was inserted into the rebuilded reporter vector pGL3-neo,the recombinant was identified by DNA sequencing,PCR and restriction enzyme digestion.The plasmid pGL4-CXCR4 was transfected into Jurkat cells,the luciferase activity in stable transfected cells was detected.Results CXCR4 promoter gene fragment was amplified by PCR.The insertion sequence of CXCR4 promoter in pGL3-neo-CXCR4 was in conformity with the GenBank reports.The luciferase activity in the pGL3-neo-CXCR4 group was 869 159.0±62 618.8,which was significantly higher that in the pGL3-neo group(39 088.4±3 867.9) and the blank group(464.2±44.0),P0.05.Conclusion Luciferase reporter vector pGL3-neo-CXCR4 containing human CXCR4 promoter can be constructed successfully.