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. 2021 Aug 31;7(9):1500–1507. doi: 10.1021/acscentsci.1c00499

Figure 4.

Figure 4

Yield of sfGFP containing five copies of pBrF ncAA when expressed in Vmax X2 versusE. colistrains. (A) Time-dependent growth curves; (B) time-dependent increase in 528 nm fluorescence; (C) OD600; (D) 528 nm fluorescence; and (E) isolated yields after 4 h. Vmax X2 and BL21 cells were transformed with pET-5XTAG-sfGFP and pEVOL-CNF, whereas Top10 and C321 cells were transformed with pET-5XTAG-sfGFP and pULTRA-CNF (Top10, C321), to induce expression of sfGFP bearing an ncAA at five positions with sfGFP (S2, D36, K101, E132, and D190). After induction, cells were grown for 4 h at 37 °C (Vmax X2, BL21, Top10, C321.ΔA.exp) or 34 °C (C321.ΔA.opt) in the presence of 0.5 mM pBrF.