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  • Protocol for tracking the inheritance patterns of proteins in live Saccharomyces cerevisiae cells using a photo-convertible fluorescent protein.

Protocol for tracking the inheritance patterns of proteins in live Saccharomyces cerevisiae cells using a photo-convertible fluorescent protein.

STAR protocols (2021-06-08)
Morgane Auboiron, Pauline Vasseur, Marta Radman-Livaja
ABSTRACT

Tracking the inheritance patterns of proteins (TrIPP) is a live-cell imaging technique used for tracking maternal protein segregation patterns between mother and daughter cells during asymmetric divisions of budding yeast. We use the photo-convertible fluorescent protein Dendra2 fused to a protein of interest (POI). Irreversible conversion from green to red fluorescence allows for parallel monitoring of old and new proteins for several generations. Single-cell quantitative image analysis of time-lapse microscopy gives synthesis and decay rates, as well as segregation patterns of the POI. For complete details on the use and execution of this protocol, please refer to Auboiron et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Threonine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Tyrosine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Arginine monohydrochloride, reagent grade, ≥98% (HPLC), powder
Sigma-Aldrich
Uracil, BioReagent, suitable for cell culture
Sigma-Aldrich
L-Isoleucine, reagent grade, ≥98% (HPLC)