肝素酶的原核表达与表达条件优化

Prokaryotic Expression and Purification of Heparinases Ⅰ

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.