Figure 3. Evaluation of RF1 as a double-edged sword in reassignment of amber code.
(A) Quantitation of RF1 repressions versus UAA incorporation and host cell growth detected by GFP fluorescence and OD600, respectively, when Y39NAEK-GFP was expressed. The values were measured relative to sgRNA0 and performed by three independent batches of cells. Error bars show one standard deviation from the mean of at least three values. (B) In-cell fluorescence assay of GFP signals in BL21(DE3) strains transformed with the representative pCCR-sgRNA plasmids that led to different extent of RF1 repression. The GFP fluorescence was measured relative to wild-type GFP (WT) value. P*** < 0.001 was determined by One-way ANOVA with Dunnett’s test to compare the expression levels in all experimental groups.