Fig. 5.
Purification of TAP1479 from E. coli/pHF5Ca and Xac amy::pHF5Ca. E. coli DH10B carrying pHF5Ca and a kanR mutant of Xac harboring pHF5Ca integrated into its chromosome were induced for the production of the TAP1479. Following expression, cells were lysed and protein extracts subjected to affinity chromatography through IgG-sepharose. (A) E. coli/pHF5Ca; and (B) Xac amy::pHF5Ca, Coomassie blue-stained 10% SDS-PAGE showing the separation of proteins from the fractions collect throughout the process. (A) lane: 1, clarified lysate; 2, flow through; 3–6, washes (20 column-volumes each); 7, protein molecular weight marker; 8–14, elution fractions. (B) Lane: 1, clarified lysate; 2, flow through; 3–5, washes; 6, protein molecular weight marker; 7–14, elution fractions. The position of the TAP1479 is marked with black arrows in both gels.