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to review the impression of multiple constraints on mutational tolerance throughout selection, we chose E. coli dihydrofolate reductase (DHFR) being a design program. DHFR is An important enzyme inside folate metabolism that decreases dihydrofolate to tetrahydrofolate and is also needed for thymidine generation. applying this exercise as the basis for an in vivo variety assay (Reynolds et al., 2011), we aimed initial to measure a mutational landscape for DHFR then to ascertain how a improve to the mobile surroundings could have an impact on the landscape. mainly because DHFR is understood to development through numerous conformational states all through catalysis (Boehr et al.

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(2020) Altered expression of an outstanding Handle protease in E. coli reshapes the in vivo mutational landscape of a design enzyme

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