Construction and identification of luciferase reporter gene vector of chicken IFN-β promoter.

To construct and identify a luciferase reporter vector containing the promoter of chicken IFN-β gene and to identify the transcriptional activity of the promoter in chicken embryo fibroblast cell line(DF-1).The promoter of chicken IFN-β gene was amplified from chicken genomic DNA by PCR and was subcloned into promoter free plasmid pGl3-basic to generate the recombinated plasmid pChIFN-β-luc.Then the reporter gene vectors were transiently transfected into DF-1 cells,transfected cells were either mock-treated or transfected with poly(I:C) and luciferase activities were measured.The promoter of chicken IFN-β gene was amplifed by PCR and the double enzyme digestion of recombined plasmid was completely correct.Luciferase reporter assay demonstrated that transcriptional activity was detected in the reporter gene vector pChIFN-β-luc and higher level of luciferase was obviously detected in DF-1 cells stimulated with the transfection of poly(I:C).Chicken IFN-β luciferase reporter vector is successfully constructed and it is an effective tool for studying the transcriptional regulatory mechanism of chicken IFN-β gene.