Create a laboratory workflow for recombinant protein expression and purification: (1) gene synthesis and codon optimization, (2) cloning into pET expression vector with His6-TEV tag, (3) transformation into BL21(DE3), (4) IPTG induction (0.5mM, 18°C, 16h), (5) cell lysis by sonication, (6) Ni-NTA affinity chromatography (wash 20mM imidazole, elute 250mM), (7) TEV protease tag cleavage, (8) reverse Ni-NTA to remove tag, (9) ion exchange chromatography (MonoQ), (10) size exclusion chromatography (Superdex 200). Show SDS-PAGE gel at key QC points.
