| 应用PCR重组技术构建大肠杆菌分泌型表达载体 |
| |
| 引用本文： | 黄仪秀,陈杰鹏,毕群,李斯明,张磊,朱圣庚. 应用PCR重组技术构建大肠杆菌分泌型表达载体[J]. 中国生物化学与分子生物学报, 1995, 11(6): 642-645 |
| |
| 作者姓名： | 黄仪秀 陈杰鹏 毕群 李斯明 张磊 朱圣庚 |
| |
| 作者单位： | 北京大学生命科学学院生物化学与分子生物学系，广东省汕头市滨制药厂 |
| |
| 摘 要： | 应用ＰＣＲ重组技术，对大肠杆菌分泌型表达载体ｐＩＮ－ⅢｏｍｐＡ进行改建，除去原载体的ＨｉｎｄⅢ位点，将信号肽序列末端两个密码子ＣＡＧＧＣＣ改为ＣＡＡＧＣＴ，从而引入一个新的ＨｉｎｄⅢ位点，并在其下游接上一段多克隆位点序列。改建后的载体可用于直接插入外源ＤＮＡ编码序列，表达产物在分泌过程中被切除信号肽而成为天然有活性的蛋白质，操作十分简便。
|
| 关 键 词： | PCR重组 信号肽序列 多克隆位点序列 分泌型表达载体 质粒构建 |
| 收稿时间： | 1995-12-20 |
Construction of a Expression and Secretion Vector in Escherichia coli by the Recombinant PCR |
| |
| Huang,Yi-Xiu,Chen,Jie-Peng,Bi,Qun,Li,Si-Ming Zhan,Lei,Zhu,Sheng-Geng. Construction of a Expression and Secretion Vector in Escherichia coli by the Recombinant PCR[J]. Chinese Journal of Biochemistry and Molecular Biology, 1995, 11(6): 642-645 |
| |
| Authors: | Huang Yi-Xiu Chen Jie-Peng Bi Qun Li Si-Ming Zhan Lei Zhu Sheng-Geng |
| |
| Affiliation: | (Department of Biochemistry and Molecular Biology, Peking University, Beijing 100871 |
| |
| Abstract: | A modified pIN-Ⅲ-ompA expression vector has been constructed by the recombinant PCR. The modification included:1)removing the Hind Ⅲ site of the original vector,2)mutating the last two codons of the ompA signal peptide sequence from CAG GCC to CAA GCT which codes for the same amino acids,and 3)inserting just downstream from the signal peptide sequence a polylinker removed XbaⅠsite. Any foreign proteins can be readily expressed and secreted using this modified vector by inserting its coding DNA between the Hind Ⅲ site and any restriction site in the polylinker. |
| |
| Keywords: | Recombinant PCR Signal peptide sequence Polylinker Expression and secretion vector Plasmid constructing |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国生物化学与分子生物学报》浏览原始摘要信息 |
|
点击此处可从《中国生物化学与分子生物学报》下载全文 |
