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Fig. 4 | Genome Biology

Fig. 4

From: Optimizing complex phenotypes through model-guided multiplex genome engineering

Fig. 4

Construction and characterization of final strain C321.∆A.opt. a Doubling time of clones isolated during construction and optimization of C321.∆A. Strain C321.∆A.opt was constructed in seven cycles of MAGE in batches of up to three cycles separated by MASC-PCR screening to pick clones with the maximum number of alleles converted (see “Methods”). The two dotted horizontal lines correspond to the relative doubling times for the original GRO and the wild-type strain. b Testing nsAA-dependent protein expression using the nsAA p-acetyl-L-phenylalanine (pAcF) in sfGFP variants with 0, 1, or 3 residues replaced with UAG codons. Normalized GFP fluorescence was calculated by taking the ratio of absolute fluorescence to OD600 of cells suspended in phosphate buffered saline (PBS) for each sample and normalizing to the fluorescence ratio of non-recoded strain EcNR1.mutS.KO expressing 0 UAG sfGFP plasmid

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