泉州师范学院学报
泉州師範學院學報
천주사범학원학보
Journal of Quanzhou Normal College
2014年
2期
20~22
,共null页
肝素酶HepI 重组 表达 纯化
肝素酶HepI 重組 錶達 純化
간소매HepI 중조 표체 순화
Heparinases I ; recombination; expression ; purification
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．coliBL21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．
利用PCR技術從肝素黃桿菌剋隆到肝素酶HepI基因，通過雙酶切將其剋隆到原覈錶達載體pGEX-4T-2中，轉化大腸桿菌E．coliBL21感受態細胞，穫得基因工程重組菌．12℃下IPTG誘導錶達12h，Glutathione Sepharose 4B純化後穫得較高純度的HepI酶蛋白．
이용PCR기술종간소황간균극륭도간소매HepI기인，통과쌍매절장기극륭도원핵표체재체pGEX-4T-2중，전화대장간균E．coliBL21감수태세포，획득기인공정중조균．12℃하IPTG유도표체12h，Glutathione Sepharose 4B순화후획득교고순도적HepI매단백．
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.
