厦门大学学报（自然科学版）2011，Vol.50Issue(1)：82-87,6.
高密度培养大肠杆菌表达人角质细胞生长因子
High-cell-density Cultivation of Escherichia coli Expressing Human Keratinocyte Growth Factor
摘要
Abstract
Keratinocyte growth factor (KGF) is the seventh member of the fibroblast growth factor family,and plays important role in the process af the proliferation of epithelial cell,the development of embryonal tissue and wound repairing. In this study, the Escherichia coli stain BL21(DE3) cloned with the plasmid pGEX-2T-KGF to expressing the fusion protein GST-KGF. The optimized conditions of induction temperature, the density of bacteria, induction time and inducer concentration for expression of fusion protein GST-KGF were investigated. When cultivating in Riesenberg synthetic medium without isopropyl β-D-thiogalactopyranoside (IPTG)induction, feedback control of substrate was applied , and the concerntration of glucose was controlled between 0.2 g/L and 0. 5 g/L,the cell dry mass concentration could reach 46 g/L. Cultivating in fermentor and introduced by IPTG in the exponential phase, the immunological activated fusion protein GST-KGF from both soluble and insoluble product were obtained by ELISA determination,108 μg/L KGF from soluble product could be achieved. An optimized process is established for production of activated human KGF by prokaryotic expression system, which lays a foundation for mass production of KGF for wound healing.关键词
角质细胞生长因子/大肠杆菌BL21(DE3)/表达/高密度培养分类
生物科学引用本文复制引用
卓山龄,孙金鹏,陈溥,傅成龙,凌雪萍,卢英华..高密度培养大肠杆菌表达人角质细胞生长因子[J].厦门大学学报（自然科学版）,2011,50(1):82-87,6.基金项目
福建省高等学校新世纪优秀人才支持计划 （）
