To study the influence of several constraints on mutational tolerance all through collection, we selected E. coli dihydrofolate reductase (DHFR) as a model method. DHFR is an essential enzyme within just folate metabolism that cuts down dihydrofolate to tetrahydrofolate and is essential for thymidine output. working with this action as The idea for an in vivo collection assay (Reynolds et al., 2011), we aimed 1st to evaluate a mutational landscape for DHFR then to find out how a adjust on the cellular atmosphere may well have an affect on the landscape. due to the fact DHFR is known to progress through a number of conformational states all through catalysis (Boehr et al.
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choice coefficients for –Lon variety (determine one—supply info 1) compared to monoculture advancement premiums measured in a plate reader in ER2566 ∆folA/∆thyA (–Lon) as described in resources and techniques.
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