生物技术通报Issue(9)：238-243,6.DOI:10.13560/j.cnki.biotech.bull.1985.2015.09.034
抗镰刀菌单链抗体在大肠杆菌中可溶性表达条件的研究
A Study on the Optimization of Condition for Soluble Expression of Fusarium-specific scFv Antibody in Escherichia coli
摘要
Abstract
The objectives of the work is to optimize the conditions for inducing expression, and obtain the soluble and high-yield expression of a Fusarium-specific single-chain variable fragment(scFv)antibody in the periplasmic space of Escherichia coli XL1-Blue. The recombinant plasmid containing a Fusarium-specific scFv antibody FvSG7 was transferred into E. coli XL1-Blue. After the culture medium for inducing selected, the soluble expression level and activity of FvSG7 antibody were analyzed by Western blot and ELISA detection for studying the influence of temperature, IPTG concentration and induction time on expression level. The maximum productivity of soluble FvSG7 antibody was obtained after induction at 25℃for 2 h, with a final concentration of 0.1 mmol/L β-D-thiogalactopyranoside(IPTG)and the cultured bacteria growing to the OD600 value of 0.5. In conclusion, the soluble expression of FvSG7 antibody in the periplasm of E. coli XL1-Blue increased significantly by optimizing the expression’s conditions of induction temperature, IPTG concentration and induction time.关键词
单链抗体/可溶性表达/大肠杆菌/优化Key words
single-chain variable fragment antibody/soluble expression/Escherichia coli/optimization引用本文复制引用
胡祖权,李和平,吴平,廖玉才,张静柏..抗镰刀菌单链抗体在大肠杆菌中可溶性表达条件的研究[J].生物技术通报,2015,(9):238-243,6.基金项目
国家重点基础研究发展计划项目（2013CB127801），国家自然科学基金项目 （）
