to review the influence of numerous constraints on mutational tolerance in the course of collection, we selected E. coli dihydrofolate reductase (DHFR) for a model procedure. DHFR is A necessary enzyme within folate metabolism that minimizes dihydrofolate to tetrahydrofolate and is particularly needed for thymidine manufacturing. Using this activity as the basis for an in vivo assortment assay (Reynolds et al., 2011), we aimed very first to evaluate a mutational landscape for DHFR and then to ascertain how a improve for the cellular natural environment could possibly impact the landscape. due to the fact DHFR is understood to progress via various conformational states throughout catalysis (Boehr et al.
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