Isolating and Engineering Fluorescence-Activating Proteins Using Yeast Surface Display.

Autor:	El Hajji L; Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, Paris, France., Benaissa H; Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, Paris, France., Gautier A; Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, Paris, France. arnaud.gautier@sorbonne-universite.fr.; Institut Universitaire de, Paris, France. arnaud.gautier@sorbonne-universite.fr.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2491, pp. 593-626.
DOI:	10.1007/978-1-0716-2285-8_25
Abstrakt:	This protocol describes the workflow to isolate and engineer fluorescence-activating proteins by yeast surface display. Fluorescence-activating proteins are an emerging class of fluorescent chemogenetic reporters for monitoring gene expression and protein localization in living cells and organisms. They become fluorescent upon binding exogenously applied fluorogenic organic dyes. Efficient fluorescence-activating proteins can be selected from yeast-displayed libraries by iterative rounds of fluorescence-activated cell sorting. The overall strategy is described, as well as a strategy for characterizing the affinity and spectroscopic properties of the selected clones. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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