ACS Synthetic Biology · 2022 · 45 citations · 56 references
Life SciencesCell BiologyMolecular BiologyCell DivisionGene ExpressionMolecular ImagingCell Growth KineticsCellular ImagingFluorescence ImagingFluorescence MicroscopyGene Expression ImagingOptogeneticsBrightness MeasurementFluorescence Imaging Of Gene ExpressionFluorescence Microscopy QuantificationIn Vivo Fluorescence ImagingIn Vivo StudiesMicroscopyOptogenetic StimulationPrecursor Protein Maturation
maturation for 12 FPs in budding yeast. To overcome the technical limitations of translation inhibitors commonly used to study FP maturation, we implemented a new approach based on the optogenetic stimulations of FP expression in cells grown under constant nutrient conditions. Combining the rapid and orthogonal induction of FP transcription with a mathematical model of expression and maturation allowed us to accurately estimate maturation rates from microscopy data in a minimally invasive manner. Besides providing a useful resource for the budding yeast community, we present a new joint experimental and computational approach for characterizing FP maturation, which is applicable to a wide range of organisms.
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Enzymatic assembly of DNA molecules up to several hundred kilobases
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A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum
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