Fig. 1: Molecular evolution of the IsPadC-PCM variants in bacteria.
From: Single-component near-infrared optogenetic systems for gene transcription regulation
a Schematics outlining the high-throughput screening of IsPadC-PCM mutants with the light-controlled behavior. b Plasmids used in the molecular evolution of IsPadC-PCM. Left: pLEVI(408)-ColE-IsPadC-PCM encodes light-sensing protein LexA408-DBD-IsPadC-PCM-msfGFP under constitutive J23116 promoter and mCherry reporter under constitutive ColE promoter controlled by LexA408 operator. Right: pWA23h-bla encodes heme oxygenase for biliverdin production under constitutive β-lactamase promoter. c Repression of the mCherry reporter expression with 660 nm light. Streaks of bacteria expressing the indicated IsPadC-PCM mutants were grown on Petri dishes either in darkness or under 660 nm light and then imaged using 570/30 nm excitation and 615/40 nm emission filters. d Efficiency of the mCherry repression by selected IsPadC-PCM variants. mCherry signal was measured in bacterial suspensions grown in darkness or under 660 nm light. Box plots show the median (center line), first and third quartiles (box edges), 1× the SD (whiskers), and individual data points. n = 4 independent experiments. a.u., arbitrary units. Source Data are available as a Source Data file.
