葡萄糖激酶的克隆表达  

Cloning and Expression of Glucokinase

作　　者：王佳宁 陈少云[1] WANG Jia-ning;CHEN Shao-yun(College of Life Science, Zhejiang Chinese Medical University, Hangzhou Zhejiang 310053)

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《海外文摘》2018年第2期84-86,共3页Overseas Digest

基　　金：中国博士后科学基金面上资助(NO.2015M581963);浙江省博士后科研项目择优资助(NO.BSH1502156).

摘　　要：目的:实现葡萄糖激酶GK的克隆表达。方法:通过酶切、酶连、转化构建重组质粒pCDFDuet-GK,进而构建工程菌;研究诱导温度、诱导时机、诱导强度、诱导时间对酶活力的影响,优化表达条件。结果:成功构建表达载体pCDFDuet-GK和工程菌,实现了葡萄糖激酶的克隆表达,在诱导温度16℃,诱导时机OD6000.6,诱导强度0.6mMIPTG,诱导时间12h的最优条件下,最大酶活力达到6.6U/L。Objective:Achieve cloning and expression of glucokinase GK. Methods: The recombinant plasmid pCDFDuet-GK was constructed through enzyme digestion, link and conversion, and used to build an engineering strain. The influence of induction opportunity, induction temperature, induction intensity and time on enzyme activity was investigated to optimize the expression conditions. Result: Expression vector pCDFDuet-GK and engineering strain were constructed successfully, and result in successful cloningand expression of glucokinase. Highest enzyme activity reached 6.6 U/L at the induction temperature of 16℃, induction opportunity of OD6000.6, induction intensity of 0.6 mM IPTG and induction time of 12 h.

关 键 词：葡萄糖激酶 克隆 表达 

分 类 号：Q552[生物学—生物化学]