Ghuge AAAnderson RAGottfried SDaube CKoloamatangi SMBMJSchiemann AHSattlegger E2024-01-102024-07-252023-01-252024-01-102024-07-252023-03-17Ghuge 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-).2666-1667https://mro.massey.ac.nz/handle/10179/70946Here, 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.(c) 2022 The Author/sCC BY-NC-ND 4.0https://creativecommons.org/licenses/by-nc-nd/4.0/Biotechnology and bioengineeringCell-based AssaysModel OrganismsMolecular BiologySaccharomyces cerevisiaeAmino AcidsSaccharomyces cerevisiae ProteinsPhenotypeJournal article10.1016/j.xpro.2022.1015452666-1667journal-article101545-https://www.ncbi.nlm.nih.gov/pubmed/36856772101545S2666-1667(22)00425-7