Optimization of an In Vitro Transcription/Translation System Based on Sulfolobus solfataricus Cell Lysate

Figure 6

SsOGT labeling. SDS-PAGE of in vitro-expressed pBS-rRNA_p-ogt plasmid and purified SsOGT protein both incubated with the BG-FL substrate (5 μM) for 60 min at 70°C. The gel was exposed for fluorescence imaging analysis, blotted, and stained with Coomassie blue. The filter was probed with the anti-OGT antibody (middle panel). Lane 1 contains 100 μg of S. solfataricus S30 fraction in the presence of the BG-FL substrate; lane 2 contains 8 μg of pBS-rRNA_p-ogt plasmid in 100 μg of S. solfataricus S30 fraction and BG-FL substrate; lane 3 contains 200 ng of purified OGT protein with 100 μg of S. solfataricus S30 fraction and BG-FL substrate; lane 4 contains 200 ng of purified OGT protein with BG-FL substrate; lane 5 contains 100 μg of S. solfataricus S30 fraction; and lane 6 corresponds to the protein marker.