人骨形成蛋白-7成熟肽在大肠杆菌中的高效表达  

Expression of mature peptide of human bone morphogenetic protein-7 in escherichia coli

作　　者：王琦[1] 葛宝丰[1] 殷莹[1] 白孟海[1] 高梅[1] 

机构地区：[1]兰州军区兰州总医院骨科研究所,甘肃兰州730050

出　　处：《中国骨伤》2005年第12期737-739,共3页China Journal of Orthopaedics and Traumatology

摘　　要：目的:利用大肠杆菌高效表达人骨形成蛋白-7(human bone morphogenetic protein-7,hBMP-7)成熟肽.方法:将编码hBMP-7成熟肽cDNA的基因片段克隆入受控于PRPL启动子的温控型大肠杆菌表达载体pDH,构建成的重组质粒pDHB-7 m以大肠杆菌DH5a为宿主菌进行温度诱导表达.结果:含重组质粒的工程菌经42℃诱导表达后在SDS-聚丙烯酰胺凝胶电泳上显现出一条新蛋白区带,分子量约为16 ku,表达量占菌体总蛋白的20%～25%,以包涵体形式存在,经简单纯化处理,得到纯度高于80%的人骨形成蛋白-7成熟肽.结论:hBMP-7成熟肽在大肠杆菌中得到高效表达,为深入研究其生物学活性和临床应用奠定了基础.Objective： To study the expression of the mature peptide of human bone morphogenetic protein-7 （hBMP-7） in escherichia coll. Methods：The cDNA fragment encoding the mature peptide of hBMP-7, with start codon and two tandem stop codons,was inserted into expression vector pDH in which fordgn gene was controlled by PRPL promoters. The recombinant plasmid pDHt3-7m was transformed into E. coli DHSa and induced at 42 ℃ to express the encoded protein. Results：After induction, a new anticipated 16 ku protein band appeared on SDS-PAGE gel and amounted to 20% to 25% of total bacterial protein. The expressed product existed in a form of inclusion body. After being partially purified and refolded, rhBMP-7m could heterotopically induce the formation of cartilage and bone tissue. Conclusion： The mature peptide of hBMP-7 has been successfully expressed in E. coli, and it lays the foundation for the study of biological activity and clinical practice.

关 键 词：基因表达 DNA 重组 大肠杆菌 

分 类 号：R692[医药卫生—泌尿科学] R378.21[医药卫生—外科学]