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Title: Biological fitness landscapes by deep mutational scanning
Knowledge of the distribution of fitness effects (DFE) of mutations is critical to the understanding of protein evolution. Here, we describe methods for large-scale, systematic measurements of the DFE using growth competition and deep mutational scanning. We discuss techniques for producing comprehensive libraries of gene variants as well as provide necessary considerations for designing these experiments. Using these methods, we have constructed libraries containing over 18,000 variants, measured fitness effects of these mutations by deep mutational scanning, and verified the presence of fitness effects in individual variants. Our methods provide a high-throughput protocol for measuring biological fitness effects of mutations and the dependence of fitness effects on the environment.  more » « less
Award ID(s):
1817646
PAR ID:
10339728
Author(s) / Creator(s):
;
Date Published:
Journal Name:
Methods in enzymology
Volume:
643
ISSN:
0076-6879
Page Range / eLocation ID:
203-224
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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