中国组织工程研究2012，Vol.16Issue(24)：4505-4508,4.DOI:10.3969/j.issn.1673-8225.2012.24.028
重组pcDNA4His/Max人骨形态发生蛋白9真核表达载体的构建与鉴定
Construction and identification of a recombinant pcDNA4 His/Max human bone morphogenetic protein eukaryotic expression vector
江维 1胡侦明 2郝杰 2甘强1
作者信息
- 1. 重庆医科大学,重庆市,400016
- 2. 重庆医科大学附属第一医院骨科,重庆市,400016
- 折叠
摘要
Abstract
BACKGROUND: Adenovirus mediated method has been used numerously in human bone morphogenetic protein 9 (hBMP-9) induced osteogenesis researches, but the security is the most important problem. Eukaryotic expressing vector is a way to solve this question.OBJECTIVE: To reconstruct a recombinant DNA pcDNA4 His/Max hBMP-9 eukaryotic expressing vector by amplifying hBMP9 gene and then cloning into pcDNA4 His/Max.METHODS: Padtrack-cmv-hBMP-9 was amplified by PCR, and hBMP-9 gene was retrieved by electrophoresis. pcDNA4 His/Max was digested by Not I , Kpn I and then the hBMP-9 gene was cloned into pcDNA4 His/Max. The recombinant plasmid was transformed by DH5α, clonal expansion, and purification, and then verified by sequencing.RESULTS AND CONCLUSION: The cloned hBMP-9 gene was 1.3 kb long, having the same length and sequence of the gene that human possessed. Eukaryotic expressing vector of pcDNA4 His/Max hBMP-9 has been constructed successfully.关键词
人骨形态发生蛋白9/真核表达载体/基因/腺病毒/阳性克隆分类
医药卫生引用本文复制引用
江维,胡侦明,郝杰,甘强..重组pcDNA4His/Max人骨形态发生蛋白9真核表达载体的构建与鉴定[J].中国组织工程研究,2012,16(24):4505-4508,4.
