Extended Data Fig. 9: Locked thiC intragenic inverton construction and growth competition.
From: Intragenic DNA inversions expand bacterial coding capacity
(A) Generation of locked intragenic invertons. The forward and locked forward thiC inverted repeat (IR) nucleotide sequences are shown. When possible, the wobble position of each codon corresponding to the IR was mutated to increase mismatches between the two palindromic sequences while maintaining the amino acid sequence. (B) Mutated nucleotides are highlighted in grey. (C) Locked thiC strains were competed against each other in thiamine-containing media in a 1:1 ratio. After 40 h, the abundance of each strain was enumerated using selective agar. Black bars indicate the locked forward strain and white bars indicate the locked reverse strain. Recovered abundances shown here correspond with the competitive index shown in Fig. 4d. Left - the locked forward strain is marked with an erythromycin resistant cassette and the locked reverse strain is marked with a tetracycline resistant cassette. Right - the locked forward strain is marked with a tetracycline resistant cassette and the locked reverse strain is marked with an erythromycin resistant cassette. Geometric mean and geometric standard deviation are shown. Each dot represents an individual replicate. Experiments were done in biological duplicate or triplicate and repeated 4 or 6 times. A two-tailed ratio paired t test was performed on the locked forward and locked reverse abundances. ***, p < 0.001; **, p < 0.01; *, p < 0.05.
