| 大肠杆菌葡萄糖脱氢酶基因的克隆与原核表达 |
| |
| 引用本文： | 韩增叶,孙继国,葛喜珍,田平芳.大肠杆菌葡萄糖脱氢酶基因的克隆与原核表达[J].北京化工大学学报(自然科学版),2012,39(1):68-71. |
| |
| 作者姓名： | 韩增叶 孙继国 葛喜珍 田平芳 |
| |
| 作者单位： | 北京化工大学生命科学与技术学院,北京,100029;北京联合大学生物化学工程学院,北京,100023 |
| |
| 基金项目： | 国家自然科学基金(21076013) |
| |
| 摘 要： | 克隆出大肠杆菌编码葡萄糖脱氢酶(PQQGDH)的 gcd 基因，构建了诱导型表达载体pET28a-gcd，转化大肠杆菌E. coli BL21后获得阳性克隆菌株BL21/pET28a-gcd。IPTG诱导后，经SDS-PAGE分析表明，该工程菌PQQGDH的表达量约为对照菌的18倍，约为18.2 mg/L，实现了高表达。此外，研究发现添加MgCl2能提高PQQGDH的表达量。
|
| 关 键 词： | 葡萄糖脱氢酶 吡咯喹啉醌 gcd基因 传感器 |
| 收稿时间： | 2011-04-18 |
Cloning and prokaryotic expression of glucose dehydrogenase from Escherichia coli |
| |
| HAN ZengYe,SUN JiGuo,GE XiZhen,TIAN PingFang.Cloning and prokaryotic expression of glucose dehydrogenase from Escherichia coli[J].Journal of Beijing University of Chemical Technology,2012,39(1):68-71. |
| |
| Authors: | HAN ZengYe SUN JiGuo GE XiZhen TIAN PingFang |
| |
| Affiliation: | 1.College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029;2.Biochemical Engineering College of Union University, Beijing 100023, China |
| |
| Abstract: | The gcd gene-encoding pyrroloquinoline quinine(PQQGDH) has been cloned using the polymerase chain reaction(PCR) from E.coli BL21,and an inducible expression vector pET28a-gcd was constructed and transformed into E.coli BL21.Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) analysis showed a high expression of PQQGDH upon isopropyl β-D-1-thiogalactopyranoside(IPTG) induction(18.2 mg/L),corresponding to a 18-fold increase compared with that in control strain.Addition of MgCl2 further enhanced the expression of PQQGDH. |
| |
| Keywords: | glucose dehydrogenase pyrroloquinoline quinine gcd sensor |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《北京化工大学学报(自然科学版)》浏览原始摘要信息 |
|
点击此处可从《北京化工大学学报(自然科学版)》下载全文 |
