蚓激酶成熟肽的原核表达及纯化  

Expression and Purification of Lumbrokinase Mature Peptide Gene in E.coli

作　　者：李清竹[1] 赵玉军[2] 张守峰[1] 刘烨[2] 寇叙[3] 扈荣良[1] 

机构地区：[1]中国人民解放军军事医学科学院军事兽医研究所,吉林长春130062 [2]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [3]吉林农业大学动物科技学院,吉林长春130118

出　　处：《现代畜牧兽医》2006年第2期43-45,共3页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：国家"863"高技术研究发展计划(2002AA206653)基金资助。

摘　　要：应用PCR技术扩增获得蚓激酶成熟肽基因,并按相应的阅读框架克隆入表达载体pGEX-4T中谷胱甘肽S-转移酶(GST)的下游。重组质粒转化大肠杆菌Rosetta株,分别在1.0mmol/LIPTG、37℃和0.1mmol/LIPTG、28℃条件下诱导,蚓激酶成熟肽-GST融合蛋白获得了高效表达,且后者得到了可溶性蛋白。经SDS-PAGE证实所表达的融合蛋白产物相对分子量与预期的53kD相符。并且以等电点和亲和层析的方法对表达出的可溶性蛋白进行了纯化。The mature peptide DNA of Lumbrokinase was amplified by PCR and subcloned downstream the glutathione stransferase （GST） by the exactitude open reading frame of vector pGEX-4T. The recombinant plasmid was transformed into E·coli. Rosetta and then inducted by two conditions respectively： 1.0 mmol/L IPTG AT 37℃ and 0.1 mmol/L IPTG AT 28℃. The fusion protein of GST-Lumbrokinase was high yield expressed. The dissolvable protein could be obtained at the latter conditions and purified by isoelectric point separation or affinity chromatograph.

关 键 词：蚓激酶 成熟肽 原核表达 纯化 

分 类 号：Q555.7[生物学—生物化学]