大肠杆菌葡萄糖脱氢酶基因的克隆与原核表达
Cloning and prokaryotic expression of glucose dehydrogenase from Escherichia coli
韩增叶 1孙继国 1葛喜珍 2田平芳1
作者信息
- 1. 北京化工大学生命科学与技术学院,北京100029
- 2. 北京联合大学生物化学工程学院,北京100023
- 折叠
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/pET28 a-gcd.IPTG诱导后,经SDS-PAGE分析表明,该工程菌PQQG-DH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.
Abstract
The gcd gene-encoding pyrroloquinoline quinine ( PQQGDH) has been cloned using the polymeraae 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 MgC12 further enhanced the expression of PQQGDH.
关键词
葡萄糖脱氢酶/吡咯喹啉醌/gcd基因/传感器Key words
glucose dehydrogenase/pyrroloquinoline quinine/gcd/sensor引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据