人骨形成蛋白4成熟肽cDNA的克隆及其在大肠杆菌中的表达  

Cloning of mature peptide cDNA gene of human bone morphogenetic protein 4 and its expression in Escherichia coli

作　　者：秦云[1] 陈苏民[1] 关路媛[1] 陈南春[1] 张立藩[2] 

机构地区：[1]第四军医大学基础部生物化学与分子生物学教研室 [2]第四军医大学航空航天医学系航空航天生理学教研室,陕西西安710033

出　　处：《第四军医大学学报》2005年第3期196-198,共3页Journal of the Fourth Military Medical University

摘　　要：目的:克隆编码人骨形成蛋白4(hBMP4)成熟肽的 cDNA基因,并在大肠杆菌中表达.方法:从人胎盘组织中提 取总RNA,以其为模板,采用RT PCR方法得到编码hBMP4 的成熟肽段(hBMP4m)cDNA,克隆入表达载体pDH2中,转 化大肠杆菌DH5α,温度诱导表达.结果:获得hBMP4mcDNA基因,成功克隆入温度诱导表达载体pDH2,DNA序列分析证 实所插入的hBMP4mcDNA序列与设计预期一致;将获得重 组表达质粒pDH2 hBMP4m转化大肠杆菌DH5α中,经温度诱 导,目的蛋白占细菌总蛋白的41.5%.结论:采用分子克隆的 方法得到编码hBMP4成熟肽段的cDNA,克隆入表达载体,在 大肠杆菌中成功获得hBMP4成熟肽的高效表达.AIM: To clone the cDNA coding for human bone morphogenetic protein 4 mature peptide (hBMP4m) and to express the peptide in Escherichia coli. METHODS: Total RNA from human placenta was extracted and the cDNA coding for hBMP4m was obtained by RT-PCR using placenta total mRNA as the template. The cDNA was cloned into expression vector pDH2 and transformed to Escherichia coli DH5α. After the sequence of the inserted gene was confirmed as designed, pDH2-hBMP4m/DH5α was induced at 42℃. RESULTS: Mature peptide cDNA coding for hBMP4m was obtained and was proven to have the sequence as designed. The expression vector pDH2-hBMP4m was constructed. After the vector was transformed into DH5α and induced at 42℃, the target protein, hBMP4m, accounted for 41.5% of the total bacterial protein. CONCLUSION: We have successfully cloned the cDNA coding for hBMP4m and constructed the vector capable of expressing hBMP4m at a high level.

关 键 词：BMP 成熟肽 人骨形成蛋白 克隆 大肠杆菌 诱导表达 DH5Α DNA 表达载体 高效表达 

分 类 号：R378[医药卫生—病原生物学] R780.2[医药卫生—基础医学]