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. 2009 Dec 3;38(4):1217–1227. doi: 10.1093/nar/gkp1079

Figure 5.

Figure 5.

Five percent denaturing polyacrylamide gels of in vitro coupled transcription/translation reactions expressing 35S-labeled proteins. (A) Lane 1, no DNA template. Lane 2, positive control test plasmid encoding the 32 kDa elongation factor EF-Ts protein. Lane 3, polycistronic mRNA generated in vitro from pCZB5 plasmid showing CT α (37 kDa) and CT β (33 kDa). Lane 4, same as lane 3 with the addition of 166nM unlabeled wild-type CT to the transcription/translation reactions. Lanes 5–15 contain 1 µg (∼10 nM) pCZB5 plasmid. Lanes 5–10 contain 0, 63, 125, 250, 500 and 1000 nM unlabeled wild-type CT. Lanes 10–15 contain 1 µM unlabeled wild-type CT and acetyl-CoA at 0, 88, 175, 350, 700 and 1400 µM. (B) Lane 1, no DNA template. Lane 2, positive control test plasmid showing a 32kDa protein. Lane 3–11 contain 1 µg (∼10nM) pGLW39 plasmid. Lanes 3–7 contain 0, 63, 125, 250 and 1000 nM unlabeled wild-type CT. Lanes 7–11 contain 1 µM unlabeled wild-type CT and acetyl-CoA at 0, 175, 350, 700 and 1400 µM. (C) Lane 1, no DNA template. Lanes 2–8 contain positive control test plasmid as in (A), lane 2. Lanes 3–8 contain 31, 63, 125, 250, 500 and 1000 nM unlabeled wild-type CT.