FIG. 2.
Denaturing SDS-PAGE gel showing purified DoxA. Lane 1, protein molecular size standards (in kilodaltons) as follows: lysozyme, 14; β-lactoglobulin, 18; carbonic anhydrase, 29; ovalbumin, 43; bovine serum albumin, 68; phosphorylase B, 97; and myosin (heavy chain), 200. Lane 2, S. lividans TK24(pANT195) crude extract. Lane 3, the 30 to 60% ammonium sulfate precipitation concentrate. Lane 4, pooled Q-Sepharose Fast Flow (anion exchange) fractions. Lane 5, pooled Hi-S (cation exchange) fractions. Lane 6, pooled phenyl Sepharose CL-4B (hydrophobic interaction) fractions. Lane 7, the fraction containing purified DoxA from Superose 6 (HPLC gel filtration). Purified DoxA was obtained after Superose 6 gel filtration (denoted by the arrow). The approximate Mr as determined from the SDS-PAGE gel is 47,000.