文献类型：专利

英文题名：Production of purified tumor metastasis suppressor protein GDI2 by performing shake culture of single fresh Escherichia coli engineered monoclonal bacteria, adding isopropyl-beta-D-1-thiogalactopyranoside, centrifuging, and purifying

作者：DONG Y;NI K;SUN A;WANG L;WANG X;WEI D;XU R

机构：[1]UNIV EAST CHINA SCI&TECHNOLOGY

申请号：CN102286581-A

申请日：2011-07-04

公开日：2011-12-21

语种：英文

收录：DERWENT

摘要：NOVELTY - Purified tumor metastasis suppressor protein GDI2 is produced by performing shake culture of a single fresh Escherichia coli engineered monoclonal bacteria in Luria-Bertani (LB) culture medium containing ampicillin at 37 degrees C, inoculating and culturing it in optimized LB culture medium; adding isopropyl- beta -D-1-thiogalactopyranoside (IPTG) to induce at 37 degrees C, and continuously culturing for 6 hours; and centrifuging and collecting cell, adding lysis buffer solution, collecting supernatant fluid, purifying target protein, and collecting liquid to obtain the purified protein GDI2. USE - Method of producing purified tumor metastasis suppressor protein GDI2 (claimed). ADVANTAGE - The method has simple purifying process, high product purity, easy production and low cost, and is suitable for industrialization. DETAILED DESCRIPTION - Production of purified tumor metastasis suppressor protein GDI2 comprises: (A) performing shake culture of a single fresh Escherichia coli engineered monoclonal bacteria in Luria-Bertani (LB) culture medium containing ampicillin at 37 degrees C and 200 revolutions/minute (rpm) overnight, and then inoculating it in optimized LB culture medium and continuously culturing under the same condition; (B) when optical density (OD) at 600 degrees C is 0.6-0.8, adding IPTG to induce at 37 degrees C, and continuously culturing at 200 revolutions/minute for 6 hours; and (C) centrifuging and collecting cell, adding lysis buffer solution, ultrasonically crushing, collecting supernatant fluid, purifying target protein through Profinity exact fusion-tag system (RTM: affinity tag-based protein purification system) purification column by buffer washing the purification column, passing supernatant through the column, then using upper part wash buffer washing column hybrid protein, adding elution buffer, incubating for 1 hour, and collecting liquid to obtain the purified protein GDI2.