FIG. 3.

Galactosyl transferase activity of Rv3782. Reaction mixtures contained 1 μCi of UDP-[¹⁴C]Galp, 20 μM UDP-GlcNAc, 20 μM TDP-Rha, membranes (1.5 mg), and cell envelope (1.5 mg) of control (M. smegmatis mc²155/pVV2) and overproducing (M. smegmatis mc²155/pVV2-Rv3782) strains and buffer A in a final volume of 320 μl. Following incubation, lipids were extracted and 10% aliquots were loaded onto a TLC plate and separated in CHCl₃-CH₃OH-NH₄OH-1 M ammonium acetate-H₂O (180:140:9:9:23). The TLC plate was exposed to the autoradiography film for 5 days at −70°C. Lanes: 1, glycolipids 1 to 5 radiolabeled with UDP-[¹⁴C]GlcNAc (600 dpm); 2, [¹⁴C]Gal-labeled lipids from the control (300 dpm); 3, [¹⁴C]Gal-labeled lipids from the overproducer (600 dpm).