Abstract:

Objective  To observe and compare the luciferase activities of different length segments of human dentin matrix protein 1 promoter in human dental pulp stem cells(HDPSC), obteoblasts(OC) and Hela cells. Methods The differentlength disired DNA segments were obtained from 2 0195 bp Dmp1 promoter coloned by PCR method. The amplified promoter segments with different length were cloned into luciferase report gene vector pGL3-Basic, the correct orientation of those inserts was verified by cutting with two different restrict enzymes. The luciferase activity was observed atfer different pGL3-PDmp1 vectors were transfected transiently into those three different-type cells.Results 6Dmp1 promoter segments with different-length wre obtained successfully, and luciferase report gene vectors withdifferent promoter segments were successfully constructed after identified by restriction enzymes cutting. They had different luciferase activities when theywere transfected transiently into HDPSC, and the region of -505---193bp and -935---505bp could be regarded as the specific promoters of Dmp1 promoter for HDPSC and OC respectively, which could include the basic regulatory elements. Conclusion The correct clone of the upstream of human Dmp1 promoter segments with different length had been obtained, and they had strong luciferase activities in HDPSC and OC, but very low in Hela cell. These results will make and important basis for studying mineralized tissue-specifiec transcriptional regulation mechanisms of Dmp1.

Key words: dentalmatrixprotein1, humandentalpulpstemcells, osteoblast