Study On SNP Site Function Of Gyne In GLUT9 Gene Promoter Region

Objectives: Previous studies have shown that the G to C substitution of rs13124007 located in the putative promoter region of glucose transporter 9(GLUT9), is significantly associated with gout susceptibility in Han Chinese males. Here, we investigate the potential mechanisms underlying association between this polymorphism and gout risk.Methods: Specific fragments incorporating the GLUT9 rs13124007 polymorphism were obtained by polymerase chain reaction(PCR), and cloned into a luciferase reporter vector to construct the wild(p GL3-basic-GLUT9/G) and mutant(p GL3-basic-GLUT9/C)recombinant. The recombinant were transient transfected into 293 T cells to express.Differences in promoter activities were tested by dual-luciferase activity assay.Electrophoretic mobility shift assay(EMSA) was also performed to determine differences in transcription factor binding activities between G and C alleles.Results: By sequenced the recombinant contained both the sequence of p GL3-basic and that of rs13124007 of human GLUT9 gene promoter. The segment inserted was in the correct direction. Dual-luciferase activity assays showed significant reductions in transcriptional efficiency with the C compared with G allele(P ? 0.05). Furthermore,EMSA demonstrated that the C and G alleles have different affinities for specific transcription factors, and suggests that the G to C substitution of rs13124007 causes IRF-1 binding site loss.Conclusions: The rs13124007 polymorphism in the GLUT9 promoter effectively influences GLUT9 expression, and possibly IRF-1 plays an important role in the procession. It provides the knowledge to investigate the pathology of gout in Han Chinese males.