Study On The Function For 5' -flanking-sequence Region Of BMPR-Ⅱ Gene

Background: BMPR-2 gene is a member of the TGF-b superfamily, which active domain is transmembrane serine/threonine kinase. Originally found in the search of bone inducing substances, it has turned out that this evolutionary conserved growth factor receptor plays important roles in bone formation and development.There are many of factors controling the expression of some genes.Control in transcription is the most important. So it is fundamental to elucidate the upstream regions of the BMPR2 gene promoter.Objective:To know if the 5'-flanking sequence rengion of BMPR-II gene include the promoter sequence,so know about the limit sequence of the promoter . Our study is the base for researching further the promoter of BMPR-II gene . Methods: We use PCR to amplify two of 5'-flanking sequence rengion of BMPR-II gene which have some overlapped bases . And cloned them into pGEM-3z vector .That was the construct of pGEM-3z-So.Than it was cloned into promoterless luciferase reporter vector pGL3-Basic, constructed recombinant plasmid of pGL3-Basic-So.This recombinant plasmid contain about 5.5K sequences of 5'-flanking sequence rengion of BMPR-II gene. As it for the template , seven different up primers, and one of the common down primer , we use PCR to amplify seven sequentially deleted sequences, a series of sequentially deleted fragments-luciferase expression vectors were constructed by cloning the 5'-flanking sequence rengion of BMPR-II gene fragments into promoterless luciferase reporter vector,pGL3-Basic. The series of deleted recombinant plasmids were pGL3-Basic-S1, pGL3-Basic-S2, pGL3-Basic-S3, pGL3-Basic-S4, pGL3-Basic-S5 , pGL3-Basic-S6, pGL3-Basic-S7.