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. 1998 Aug;18(8):4612–4619. doi: 10.1128/mcb.18.8.4612

FIG. 4.

FIG. 4

Formation of protein-GMP complex by recombinant T. brucei capping enzyme. Proteins were incubated in 50 mM Tris-HCl (pH 7.5)–5 mM MgCl2–5 mM DTT–[α-32P]GTP for 10 min at 28°C, denatured, and analyzed by SDS-PAGE. An autoradiograph of the gel is shown. The sources of proteins were as follows: the reaction mixture in lane 1 contained an aliquot (about 5 μg) of a T. brucei whole-cell extract, and the reaction mixtures in lanes 2 to 5 were incubated with an aliquot of a coupled in vitro transcription-translation reaction mixture that generated the proteins illustrated graphically to the right of the autoradiograph. The asterisk indicates a background band whose intensity varied from experiment to experiment.