表达含凝血酶识别位点的rhGM-CSF融合蛋白工程菌的选定及其酶切后目的蛋白的纯化  

Selection of Recombinant E.coli Expressing rhGM-CSF Fusion Protein Containing Thrombin Restriction Site and Purification of Expressed Product

作　　者：张学述[1] 简志愚[1] 刘裕 谢茂超 黄林[1] 肖洁 周秀彬 栗克喜[1] 

机构地区：[1]卫生部成都生物制品研究所,成都610063

出　　处：《中国生物制品学杂志》1999年第2期91-93,共3页Chinese Journal of Biologicals

摘　　要：比较 4株表达含凝血酶识别位点的rhGM CSF融合蛋白工程菌的特性 ,选定用于开发的工菌株 ,其表达产物相对分子质量为 2 6 0 0 0 ,N端 110 0 0为MS2 ,C端 15 0 0 0为rhGM CSF ,较易用凝血酶消化释放出目的蛋白。用阴离子交换、疏水和凝胶过滤层析 ,或后二者结合进行可得到适宜人体应用的纯品 ,活性达到 10 7IU mg蛋白。A strain of recombinant E.coli expressing rhGM CSF fusion protein containing thrombin restriction site was selected out on the basis of comparing the characteristics of 4 strains.The molecular weight of the expressed product was 26000,and the N and T terminals of it were MS2(11000)and rhGM CSF(15000), respectively.The goal protein was released easily after the expressed product was digested with thrombin and purified by anion exchange or hydrophobic and/or gel filtration chromatographies.The result showed that the purity of it reached 10 7 IU/mg.

关 键 词：RHGM-CSF 凝血酶 融合蛋白 层析纯化 

分 类 号：Q753[生物学—分子生物学] Q78