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. Author manuscript; available in PMC: 2012 Dec 20.
Published in final edited form as: Nat Chem Biol. 2012 Jan 15;8(3):238–245. doi: 10.1038/nchembio.768

Figure 2.

Figure 2

Rationally designing TMAO-dependent conditional activity. (a) Schematic of Glycine-duplet substitution library showing representative mutations and the pattern of mutations. Two consecutive residues were substituted by Glycine-Glycine residues to obtain a comprehensive library of Glycine-duplet substitutions. (b) The mutants were grown with gentamicin selection in absence of TMAO at 37°C (LB), in presence of TMAO at 37°C (TMAO), and in absence of TMAO at 30°C (30D). Growth normalized with respect to cells transformed with wtGmr, that grew equally well in these three conditions. Mutants are sorted according to their growth in LB and growth in the other two conditions is plotted as bars for each mutant. (c) Gentamicin resistance as obtained by growing each mutant in 5 different concentrations of gentamicin (2 μg/ml, 4 μg/ml, 8 μg/ml, 16 μg/ml and 32μg/ml) in presence and absence of TMAO. Growth is shown as a colormap with increasing color density representing increase in growth. Details of GmR Glycine-duplet mutants used for this analysis are mentioned at the bottom of the panel. (See also supplemental Figure 11-17)