| 人骨形成蛋白-7成熟肽在大肠杆菌中的高效表达 |
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Received:September 15, 2004
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| 作者 | Author | 单位 | Unit | E-Mail |
| 王琦 |
WANG Qi |
兰州军区兰州总医院骨科研究所,甘肃兰州730050 |
The Institute of Orthopedics of Lanzhou Military General Hospital,Lanzhou 730050,Gansu,China |
earthandwang@yahoo.com.cn |
| 葛宝丰 |
GE Bao-feng |
兰州军区兰州总医院骨科研究所,甘肃兰州730050 |
The Institute of Orthopedics of Lanzhou Military General Hospital,Lanzhou 730050,Gansu,China |
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| 殷莹 |
YIN ying |
兰州军区兰州总医院骨科研究所,甘肃兰州730050 |
The Institute of Orthopedics of Lanzhou Military General Hospital,Lanzhou 730050,Gansu,China |
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| 白孟海 |
BAI Meng-hai |
兰州军区兰州总医院骨科研究所,甘肃兰州730050 |
The Institute of Orthopedics of Lanzhou Military General Hospital,Lanzhou 730050,Gansu,China |
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| 高梅 |
GAO Mei |
兰州军区兰州总医院骨科研究所,甘肃兰州730050 |
The Institute of Orthopedics of Lanzhou Military General Hospital,Lanzhou 730050,Gansu,China |
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| 期刊信息：《中国骨伤》2005年18卷，第12期，第737-739页 |
| DOI：doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz |
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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DH5α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。 |
| [关键词]:基因表达 DNA,重组 大肠杆菌 |
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| Expression of mature peptide of human bone morphogenetic protein-7 in escherichia coli |
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Abstract:
Objective: To study the expression of the mature peptide of human bone morphogenetic protein-7（hBMP-7） in escherichia coli.
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 foreign gene was controlled by PRPL promoters.The recombinant plasmid pDHB-7m was transformed into E.coli DH5α 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. |
| KEYWORDS:Gene expression DNA,recombinant Escherichia coli |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 王琦,葛宝丰,殷莹,白孟海,高梅.人骨形成蛋白-7成熟肽在大肠杆菌中的高效表达[J].中国骨伤,2005,18(12):737~739 |
| 英文格式: | WANG Qi,GE Bao-feng,YIN ying,BAI Meng-hai,GAO Mei.Expression of mature peptide of human bone morphogenetic protein-7 in escherichia coli[J].zhongguo gu shang / China J Orthop Trauma ,2005,18(12):737~739 |
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