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. 1998 Jul 21;95(15):8619–8624. doi: 10.1073/pnas.95.15.8619

Figure 2.

Figure 2

Normal translation in sup70–65 cells. (a–d) Isogenic diploid wild-type (strain LMDWU) and sup70–65 mutant cells (strain LMD651U) were incubated on nitrogen-rich solid complex medium at 30°C. Arrows in b indicate sup70–65 mutant colonies; unmarked colonies are wild type. In c and d the medium contained cycloheximide to slow the growth of wild-type (c) and mutant (d) cells by 50%. (e and f) CAG codons are translated efficiently in sup70–65 mutant cells. β-Galactosidase activity was measured for extracts made from sup70–65 (strain LMD651U) (closed symbols) and wild-type (strain LMDWU) cells (open symbols) proliferating at 30°C and harboring plasmids pQA1, pQA2, pQA3, pQA4, pQA6, pQA7, pQA10, pQG1, pQG2, pQG3, pQG4, pQG6, or pQG8. The plasmid-encoded β-galactosidase contains (Ser-Gln5-Arg)n inserted near the N terminus, with n indicated by the number in the plasmid name. Inserted glutamines all are encoded by CAG for pQG plasmids (e) and by CAA for pQA plasmids (f). Assays were performed in duplicate, and values normalized to protein content, expressed in Miller units (×103), were used as a measure of translation efficiency. (g) Ochre suppression. Diploid strains (MLD14, MLD15, MLD17, and MLD13) with the indicated genotypes were incubated at 30°C on tryptophan-free solid medium.