人骨形成蛋白－2AcDNA基因工程产物诱骨活性的形态学证明

MORPHOLOGICAL DEMONSTRATION OF INDUCED BONE ACTIVITY FOR GENETIC ENGINEERING PRODUCTS OF HUMAN BONE MORPHOGENETIC PROTEIN 2A

人骨形成蛋白２Ａ（ｈｕｍａｎｂｏｎｅｍｏｒｐｈｏｇｅｎｅｔｉｃｐｒｏｔｅｉｎ２Ａ，ｈＢＭＰ２Ａ）ｃＤＮＡ３′端５６７个核苷酸插入表达载体ＰＳＧ４Ｔ－２中，在大肠杆菌中获得１５％的表达，表达产物为２１ＫＤ的蛋白质，经包涵体洗涤，尿素变性和复性后，取１ｉｎｇ表达产物，植入小鼠股四头肌中，２１天后取植入区进行组织切片观察，证实这种基因工程产物具有良好的异位骨诱导活性。

A 3'-terminal 567 base pairs fragment of human bone morphogenetic protein 2A(hBMP2A) gene was inserted into expression vector pSG4T-2. The 21KD protein encodedby the fragment expressed as inclusive bodies in E. colt and occupied 15% of the total cellproteins. Purification and renaturation of the protein were performend by successively washing'the pellet, solubilizing with urea and dinting with refolding buffer. 1 mg of the partiallypurified protein was implanted in mouse muscle and histological studies were carried out 21days after the operation. The results showed that the recombinant protein had good activityon inducing ectopic bone formation.