肝素酶的原核表达与表达条件优化  

Prokaryotic Expression and Purification of Heparinases Ⅰ

作　　者：杨嫦娇 郑丽燕[1] 吴文林[1] 

机构地区：[1]泉州师范学院化学与生命科学学院,福建泉州362000

出　　处：《泉州师范学院学报》2014年第2期20-22,共3页Journal of Quanzhou Normal University

基　　金：泉州市科技局项目(2010Z57);泉州师范学院校大学生科研基金资助项目(2010DKJ06)

摘　　要：利用PCR技术从肝素黄杆菌克隆到肝素酶HepI基因,通过双酶切将其克隆到原核表达载体pGEX-4T-2中,转化大肠杆菌E.coli BL21感受态细胞,获得基因工程重组菌.12℃下IPTG诱导表达12h,Glutathione Sepharose 4B纯化后获得较高纯度的HepI酶蛋白.HepariTzases I gene was cloned from F.heparinum by PCR. After digested by Not I and Sma I,it was cloned into pGEX-4T-2 plasmid with the right reading frame sequence to construct the expression vector. This plasmid was used to transfer E.coli BI.21.After inducing with IPTG at 12 ℃ for 12 h,the fusion protein was purified with Glutathione Sepharose 4B.

关 键 词：肝素酶HepI 重组 表达 纯化 

分 类 号：Q575.12[生物学—生物化学]