Construction of human IL-15 gene promoter luciferase reporter gene vectors

Purpose To clone series of the human IL-15 gene promoter and to construct seven luciferase reporter gene vectors containing IL-15 promoter regions.Methods Seven DNA fragments of the 5′ flanking region of human IL-15 gene were isolated from genomic DNA of HaCaT cells by polymerase chain reaction(PCR).After being digested with restriction enzymes,IL-15 gene family promoters were inserted into the luciferase reporter vectors PGL3-Basic.Then the recombinant construct which contains the largest IL-15 promoter region was transiently transfected into HaCaT cells.6 hours later,cells were respectively treated with DMEM,LPS.Then the activity of luciferase was detected.Results Seven-fragments of IL-15 gene promoter were amplified by PCR,and the identification of PCR and double digestion of recombined plasmid was completely correct.The experiment of transient transfection showed that the largest IL-15 promoter region has the significant activity.Conclusion The human IL-15 gene promoter luciferase reporter gene vectors have been constructed successfully.The analysis of activity in HaCaT cells indicated that the cloned IL-15 gene family promoters contain the key cis-regulatory elements.It will be essential to further study the regulation of IL-15 expression.