鼠源性甲状旁腺激素相关蛋白1～84片段的制备及其促进骨形成的作用  

Construction and expression of a recombinant mouse PTHrP 1～84 and its role in stimulating bone formation

作　　者：王子露[1] 孙雯[1] 周熙超[1] 苗登顺[1] 

机构地区：[1]南京医科大学口腔医学研究所,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第5期597-601,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生厅科研项目(H200627)

摘　　要：目的:构建并表达鼠源甲状旁腺激素相关蛋白(parathyroid hormone related protein,PTHrP)1～84片段的载体,并检测其在促进骨形成中的作用。方法:应用RT-PCR方法体外扩增mPTHrP1～84基因,经测序后重组入原核表达载体pGEX-2TK,将构建正确的pGEX-2TK/mPTHrP1～84原核表达载体转化E.coliBL21,IPTG诱导表达。表达产物经蛋白纯化及活性鉴定,通过细胞学实验确定纯化的重组PTHrP片段在促进骨形成中的作用。结果:构建的重组表达载体pGEX-2TK/mPTHrP1～84诱导表达产物以可溶性的形式存在;纯化的mPTHrP1～84可促进小鼠的间充质干细胞向成骨方向增殖和分化。结论:构建、表达的mPTHrP1～84片段可促进骨形成。Objective：To construct and express a recombinant expression vector containing a mouse parathyroid hormone related protein（PTHrP） 1～84,and to examine the effect of the purified product on bone formation.Methods：The PTHrP1～84 gene fragment was amplified by RT-PCR and inserted into corresponding sites of expression vector pGEX-2TK,and the fusion protein expressed in E.coli BL21 induced by IPTG.After purification and identification,its roles on the proliferation and differentiation of bone marrow mysenchymal stem cells was evaluated in vitro.Results：The new recombinant expression vector pGEX-2TK /mPTHrP1 ～84 was constructed successfully.The gene recombinant product was soluble.The treatment with purified mPTHrP1～84 in bone marrow cell cultures not only significantly increased colony-forming unit-fibroblast（CFU-f）,but also significantly increased alkaline phosphotase and collagen positive CFU-f and calcified nodules.Conclusion：Results demonstrated that gene recombinant mPTHrP1～84 can stimulate bone marrow mysenchymal stem cell to proliferate and differentiate into osteoblasts.

关 键 词：甲状旁腺激素相关蛋白 骨质疏松 骨形成 

分 类 号：R681.1[医药卫生—骨科学]