Analysis of human ameloblastin promoter activity regions

LIU Xiao-ying; GAO Zhi-qin; HAN Ting-ting; GAO Yu-guang

Abstract：AIM:To construct luciferase report gene vectors with different promoter segments of human amelo blastin, and to compare the luciferase activities of different length segments of human ameloblastin in ameloblast and Hela cells. METHODS: The different -length desired promoter segments were obtained by PCR method,cloned into lucifer ase report gene vectors pGL3 - Basic, and transiently transfected into ameloblast and Hela cells. We analyzed the tran scriptional regulatory capacity of the promoter segments by detecting luciferase activity. RESULTS: We have obtained different - length promoter segments of ameloblastin gene, and the different - length promoter recombinant luciferase re porter vectors were constructed successfully by cutting with two different restrict enzymes. Different - length promoters had different activities in different type cells, and -424 -267 and -128-37 regions were the specific transcriptional regulatory district. CONCLUSION: We successfully constructed the different - length recombinant pGL3 - Basic lucif erase reporter gene vector. The transcriptional activity regions of ameloblastin gene promoter were determined initially , which will provide a foundation for further studying the transcriptional regulatory characteristics of ameloblastin gene.