Construct Recombination Of Transforming Growth Factor-β Receptor Ⅱ Promoter Luciferase Repotr Gene

one of the issues affecting denture repair. The reasons causing Alveolar bone defect containboth pathological and physiological factors, such as congenital developmental abnormalities(such as cleft lip and palate, etc.), periodontal disease, trauma, tumor and hormonal changes,etc. Alveolar bone loss will affect the stability and function of jaw, the appearance, and evenseriously impact on body health. Complete denture repair is unable to get good retentionbecause of alveolar bone defection for the patients lost of denture. Fixed denture repair cannot be beautiful as a result of alveolar bone absorption severe for the patients defected ofdenture. Implant denture patients alveolar bone must be health with a certain height andwidth before surgery. The teeth of periodontal disease patients will be lost eventually alsobecause of alveolar bone defect. Therefore, alveolar bone repair and regeneration has asignificant theoretical and clinical application value in the dental field.Transforming growth factor-β（TGF-β） is a cytokine with wild functions. Studiesdemonstrated that it could promote the bone regeneration through the following ways: suchas promoting angiogenesis, stimulating bone cell and cartilage cell proliferation anddifferentiation, regulating the function of other growth factors and so on. Transforminggrowth factor-β receptors（TGFβRs）, as the receptor of TGF-β, are the base of TGF-βactivating biological function. Among them, transforming growth factor-β receptor-Ⅱ（TGFβⅡ）is one of the TGF-β high affinity receptors, which plays an important role inpromote osteoblast proliferation and differentiation.Purpose: In this research, the key gene, transforming growth factor-β receptor-Ⅱ（TGFβⅡ） that can regulate the regeneration of alveolar bone, was selected as a targetdirection, and TGFβⅡ promoter as a target point, and then TGFβⅡpromoter luciferasereporter gene was constructed. The research provides foundation of the drug screening foralveolar bone repair and regeneration. It has a profound theoretical significance andextensive clinical application value.Method: TGFβⅡ promoter sequence was searched in NCBI database, and primerpremier5.0(primer design software) was used to design TGFβⅡpromoter primer, and then the human genome was used as the template to PCR. The amplified fragments were ligatedwith pGEM-T Easy vector and the ligation products were transformed into E.coli DH5α, forscreening positive plasmids, culturing, extraction, enzyme digestion and sequencing. Then,ligation of pGEMT-TGFβⅡp recombinant plasmid and pGL2vector after double enzymedigestion was processed with following steps, those were transformation, culturing,extraction, restriction enzyme digestion,. Then the recombinant plasmid pGL2-TGFβⅡpwas constructed successfully.Result: Human TGF β Ⅱ promoter sequence was cloned by PCR, sequencecomparison revealed that there is only one point mutation, which could be explained that thepoint mutation occurred during PCR or the template itself had had mutation already. Theconstruction of pGL2-TGFβⅡp recombinant plasmid was successfully constructed.Conclusion:1Human TGF βⅡ promoter sequence was successfully cloned.2Recombinant plasmid of pGL2-TGF β Ⅱ promoter luciferase reporter gene wassuccessfully constructed. The research has a profound theoretical significance and extensiveclinical application value. It lays the foundation for the subsequent drug screening stuty.