| 一种双顺反子表达载体的构建及应用的研究 |
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| 引用本文： | 刘新平,陈苏民,陈南春,赵忠良,柴玉波,崔有宏,薛泳涛.一种双顺反子表达载体的构建及应用的研究[J].生物化学与生物物理进展,1996,23(2):156-159. |
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| 作者姓名： | 刘新平 陈苏民 陈南春 赵忠良 柴玉波 崔有宏 薛泳涛 |
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| 作者单位： | 第四军医大学生物化学及分子生物学教研室;第四军医大学生物化学及分子生物学教研室;第四军医大学生物化学及分子生物学教研室;第四军医大学生物化学及分子生物学教研室;第四军医大学生物化学及分子生物学教研室;兰州军区军医学校生化教研室;第四军医大学生物化学及分子生物学教研室 |
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| 基金项目： | 国家“八五”科技攻关资助项目85-722-07. |
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| 摘 要： | 将表达载体pEC34中的一段寡核苷酸序列，其中包括翻译增强子序列、SD序列、终止码、起始码及两端的限制性内切酶位点，插入GST基因后，构建成双顺反子的表达载体．利用此载体表达了非融合的人骨形成蛋白2A(hBMP2A)和人骨形成蛋白3(hBMP3)C端肽段，将第一顺反子基因（GST基因）切小到原来的1／3时，则位于下游的第二顺反子基因编码的蛋白质在大肠杆菌中的表达量增加一倍。
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| 关 键 词： | 双顺反子 人骨形成蛋白基因 基因表达调控 表达载体 |
| 收稿时间： | 1995/5/15 0:00:00 |
| 修稿时间： | 9/5/1995 12:00:00 AM |
Construction and Application of A Dicistronic Expression Vector |
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| Liu Xinping,Chen Sumin,Chen Nanchun,Zhao Zhongliang,Chai Yubo,Cui Youhong and Xue Yongtao.Construction and Application of A Dicistronic Expression Vector[J].Progress In Biochemistry and Biophysics,1996,23(2):156-159. |
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| Authors: | Liu Xinping Chen Sumin Chen Nanchun Zhao Zhongliang Chai Yubo Cui Youhong and Xue Yongtao |
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| Institution: | Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University;Department of Biochemistry and Molecular Biology, The Fourth Military Medical University |
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| Abstract: | A dicistronic expression vector in E.coli has been constructed. The vector contains glutathione S-transferase (GST) gene as the first cistron, followed successively with translational enhancer, SD sequence, stop codon, start codon and multiple restriction enzyme sites for cloning(MCS).3'-terminal framgents of human bone morphogenetic protein (hBMP)gene 2A and 3 were inserted into the MCS respectively. After induction, un fused hBMP2A and hBMP3 expressed and occupied 10% and 15% of the total cell protein respectively. The GST gene in the plasmids were further shortened from 660 by to 206 bp. The expression level of hBMP2A and hBMP3 were double by the plasmids containing short GST gene as compared to that of the corresponding plasmids with large GST gene. |
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| Keywords: | dicistron human bone morphogenetic protein gene gene expression regulation expression vector |
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