重组人骨形成蛋白成熟肽-4对辐射小鼠骨髓造血损伤修复作用初探

发布时间：2018-02-01 01:29

本文关键词：重组人骨形成蛋白4成熟肽重组人骨形成蛋白2成熟肽恒溶氧高密度发酵蛋白质的大肠杆菌表达蛋白质纯化辐射损伤 rhBMP-4m CD34 脾体比造血系统存活率　出处：《第四军医大学》2006年硕士论文　论文类型：学位论文

【摘要】：骨形成蛋白(Bone Morphogenetic Proteins，BMPs)是一种分泌型糖蛋白，具有多种生物学功能。BMP2和BMP4是骨形成蛋白家族中的两个重要成员，在结构和功能上这两种蛋白都非常相似，两者都能够诱导骨组织的生成和造血组织的分化等，但BMP2诱导骨组织形成的活性更强，BMP4则对造血组织的作用更突出。在成体，BMP2诱导异位骨形成，成为用于骨组织工程的重要诱导因子：BMP4则可以促进造血干细胞的增殖，并诱导造血干细胞向红系、粒系和巨噬细胞系三系细胞分化，从而能够促进受损伤的造血系统的全面再生与恢复；这些特性使得BMP2和BMP4都具有很好的临床应用前景。在BMP的制备过程中，真核表达系统表达量低，表达产物回收、纯化困难、生产成本很高；原核表达系统，特别是大肠杆菌系统，具有表达效率高，容易操作，容易监控，产物稳定，生产成本低等优点，故而我们在进行BMP的大规模制备过程中选择了大肠杆菌表达系统。目前，重组人骨形成蛋白2的大规模制备已有较多报道，，本研究的目的，就是应用我们目前已经比较成熟的制备BMP2的工艺，来制备BMP4，来验证大规模制备这两种蛋白时，能否用相同的发酵程序及相近的纯化方法。两种工程菌株(DH5α／pDH2-BMP-4m及DH5α／pDH2-BMP-2m)分别
[Abstract]:Bone morphogenetic protein (BMP) is a secretory glycoprotein. BMP2 and BMP4 are two important members of the bone morphogenetic protein family. They are very similar in structure and function. Both of them could induce bone tissue formation and hematopoietic differentiation, but BMP2 induced bone tissue formation activity was stronger. BMP4 had more effect on hematopoietic tissue, especially in adult. BMP2 induces ectopic bone formation and becomes an important inducing factor for bone tissue engineering: BMP4 can promote the proliferation of hematopoietic stem cells and induce hematopoietic stem cells to erythrocytes. The differentiation of granulocyte and macrophage cells can promote the regeneration and recovery of the damaged hematopoietic system. These characteristics make both BMP2 and BMP4 have good clinical application prospects. In the preparation of BMP, the eukaryotic expression system has the advantages of low expression level, difficult recovery of the expressed product, and high production cost. Prokaryotic expression system, especially Escherichia coli system, has the advantages of high expression efficiency, easy operation, easy monitoring, stable product, low production cost and so on. Therefore, we chose the E. coli expression system in the large-scale preparation of BMP. At present, the large-scale preparation of recombinant human bone morphogenetic protein 2 has been reported, the purpose of this study. It is to use the process of preparing BMP2, which is already mature at present, to prepare BMP4, to verify whether the two proteins can be prepared in the same fermentation process and similar purification methods. Two engineering strains, DH5 伪 / pDH2-BMP-4m and DH5 伪 / pDH2-BMP-2m, respectively
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R341

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