FIG. 2.

Complementation of PAPS-deficient E. coli with mll7575 and mll7576. (A) Colony formation on minimal medium. The cysN mutant (strain DM63, lower left), DM63 harboring vector (lower right), DM63 harboring pGTO101 (which encodes ORFs mll7575 and mll7576) (upper right), or wild-type E. coli (upper left) was grown on solid M9 minimal medium for 48 h at 37°C. (B) PAPS biosynthesis. Wild-type E. coli (lane 1), the cysN mutant (lane 2), or the cysN mutant harboring either empty vector (lane 3) or pGTO101 (lane 4) was cultured in minimal medium in the presence of Na₂³⁵SO₄. PAPS was extracted and fractionated by thin-layer chromatography, and incorporation of ³⁵SO₄ was visualized by autoradiography as described in Materials and Methods.