Figure 5 | Scientific Reports

Figure 5

From: Prokaryotic soluble overexpression and purification of oncostatin M using a fusion approach and genetically engineered E. coli strains

Figure 5

SDS-PAGE analysis of OSM purification from solubly expressed MBP-OSM and His6-OSM in E. coli. (A) MBP fusion-derived OSM purified from BL21(DE3) cells by IMAC. M, molecular weight markers; lane 1, total cell extract before IPTG induction as a negative control; lane 2, total cell extract with IPTG induction; lane 3, soluble fraction after cell sonication; lane 4, MBP-OSM fusion protein (65.95 kDa) purified by IMAC; lane 5, MBP tag cleavage reaction mixture: TEV protease (28.6 kDa), MBP tag (43.9 kDa) and OSM (22.05 kDa); lane 6, IMAC purification of OSM after TEV cleavage; lanes 7 and 8, the final OSM product under reducing and non-reducing conditions, respectively. (B) His6 fusion-derived OSM purified from Origami 2 cells by IMAC. M, molecular weight markers; lane 1, total cell extract before IPTG induction (a negative control); lane 2, total cell extract with IPTG induction; lane 3, soluble fraction after cell sonication; lane 4, His6-OSM fusion protein (25.65 kDa) purified by IMAC; lane 5, His6 tag cleavage reaction mixture: TEV protease (28.6 kDa), His6 tag (3.6 kDa) and OSM (22.05 kDa); lanes 6 and 7, the final OSM product under reducing and non-reducing conditions, respectively. Arrows indicate the positions of OSM monomers (22.05 kDa), dimers (44.1 kDa) and oligomers (≥66.15 kDa).

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