Activity analysis of promoter from human α2(I) procollagen gene
Objective To investigate the fragment of high promoter activity in human α2(Ⅰ) procollagen gene and to explore the potential effect of cytokines on promoter activity. Methods Five chimeric genes were constructed with various lengths of sequences between 2.4kb upstream of the initiation of transcription of the human α2(Ⅰ) procollagen gene and 58bp downstream of this site. The sequences were fused to chloramphenicol acetyltransferase(CAT) reporter gene. These recombinant plasmids were transiently transfected into human skin fibroblast by FuGENE 6 transfection method. The putative promoters activities were tested and compared by CAT measurement in transfected cells. Results The highest CAT expression was demonstrated in construction driven by -2292 to +58bp, -1476 to +58bp fragment. TGF-β, IGF-1 increased promoter activity of human α2(Ⅰ) procollagen gene. Conclusion TGF-β, IGF-1 increasedⅠcollagen synthesis by regulating promoter element in procollagen gene.