Rapid yeast-based screen for Functionally Relevant Amino Acids (RS-FRAA) in a protein
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Date
2023-03-17
Open Access Location
Journal Title
Journal ISSN
Volume Title
Publisher
Elsevier Inc
Rights
(c) 2022 The Author/s
CC BY-NC-ND 4.0
CC BY-NC-ND 4.0
Abstract
Here, we describe a fast and cost-effective procedure to generate a large array of mutant proteins and immediately screen for those with altered protein function. This protocol is a modification from three existing approaches: fusion PCR, Saccharomyces cerevisiae in-yeast recombination, and semi-quantitative growth assays. We also describe a mating step to reduce the occurrence of false positive findings due to ectopic mutations. The only requirement is that the protein elicits a phenotype in Saccharomyces cerevisiae.
Description
Keywords
Biotechnology and bioengineering, Cell-based Assays, Model Organisms, Molecular Biology, Saccharomyces cerevisiae, Amino Acids, Saccharomyces cerevisiae Proteins, Phenotype
Citation
Ghuge AA, Anderson RA, Gottfried S, Daube C, Koloamatangi SMBMJ, Schiemann AH, Sattlegger E. (2023). Rapid yeast-based screen for Functionally Relevant Amino Acids (RS-FRAA) in a protein.. STAR Protoc. 4. 1. (pp. 101545-).