Article
Rapid yeast-based screen for Functionally Relevant Amino Acids (RS-FRAA) in a protein.
STAR protocols - 17 Mar 2023
Ghuge Aditi A, Anderson Reuben A, Gottfried Susanne, Daube Clément, Koloamatangi Siaosi M B M J, Schiemann Anja H, Sattlegger Evelyn
Abstract excerpt
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...
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