激光生物学报2008，Vol.17Issue(3)：384-390,7.
几丁质酶基因表达载体pET-chiB的构建及其在大肠杆菌中的诱导表达
Construction of the Plasmid pET-chiB Carrying Chitinase Gene and Inducing Expression in E.coli
摘要
Abstract
Chitinase plays important roles in decomposing chitin to GIcNAc. Many chitinase genes have been separated and cloned from different kinds of microbes. In this study, the chitinase chiB gene cloned from Serrat/a marcescens was inserted into the recombinant expression plasmid pET-22b( + ) vectors at the EcoRI site, and the expression vector pET-chiB was transformed into E. coli BL21 ( DE3 ). The expression vector pET-chiB was induced to express and analyzed by SDS-PAGE, and the results indicated that its expression protein was soluble and the protein molecular weight was about 52 kD. The analysis of inducing expression was done by SDS-PAGE. The vectar'pET-chiB was induced to express different time with difference, Isopropyl-β-D-thiogalactopyranoside (IPTG) concentrations and temperatures, and the resuh showed that the best inducing parameters were 4 hours, 0.5 mmol/L and 25℃ respectively. These results provided fundamental data, and materials for the further study on chitinase gene and the producing of chitinase project strains.关键词
几丁质酶基因/pET-chiB/构建/表达/IPTG诱导Key words
chitinase gene / pET-chiB / construction/ expression / IPTG inducing分类
生物科学引用本文复制引用
叶辉,冯春,程郢,朱苏文,程备久..几丁质酶基因表达载体pET-chiB的构建及其在大肠杆菌中的诱导表达[J].激光生物学报,2008,17(3):384-390,7.基金项目
Seientifical Item of Education Department of Anhui Province ( No. 2006KJ213 B) （ No. 2006KJ213 B）
