泉州师范学院学报
泉州師範學院學報
천주사범학원학보
JOURNAL OF QUANZHOU NORMAL COLLGEG
2014年
2期
20-22
,共3页
肝素酶 HepI%重组%表达%纯化
肝素酶 HepI%重組%錶達%純化
간소매 HepI%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 ．coli BL21感受态细胞,获得基因工程重组菌．12℃下 IPTG 诱导表达12 h, 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 ．coli BL21感受態細胞,穫得基因工程重組菌．12℃下 IPTG 誘導錶達12 h, Glutathione Sepharose 4B 純化後穫得較高純度的 HepI 酶蛋白．
이용 PCR 기술종간소황간균극륭도간소매 HepI 기인,통과쌍매절장기극륭도원핵표체재체pGEX-4T-2중,전화대장간균 E ．coli BL21감수태세포,획득기인공정중조균．12℃하 IPTG 유도표체12 h, Glutathione Sepharose 4B 순화후획득교고순도적 HepI 매단백．
Heparinases 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 BL21.After inducing with IPTG at 12 ℃for 12 h,the fusion protein was purified with Glutathione Sepharose 4B.
