Our qualitative observation by wide-field (WF) microscopy suggested that, although CU17S did not fluoresce brightly in E. coli or mammalian cells, the fluorescence did not photobleach substantially. However, because it is generally assumed that there is an inverse correlation between the brightness and photostability of FPs, CU17S was not expected to improve brightness while maintaining excellent photostability. Nevertheless, through random mutagenesis of the CU17S gene, we discovered that the V168A mutation effectively improved the efficiency of both protein expression and chromophore maturation of the FP; CU17S/V168A was abundantly produced in bacteria.
Hirano et al. (2022)
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