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Figure 2 | BMC Biology

Figure 2

From: Evolution favors protein mutational robustness in sufficiently large populations

Figure 2

Outline of the neutral evolution experimental procedure. For the polymorphic population, error-prone PCR was used to generate mutant P450 genes. These genes were ligated into a plasmid and transformed into E. coli. Individual mutants (435) were picked, expressed in E. coli, and assayed for enzymatic activity. All mutants that met the selection criterion contributed an equal amount of plasmid DNA as template for the next generation of error-prone PCR. The monomorphic populations were treated similarly, except only a single mutant was assayed at each generation. If this mutant met the selection criterion then it became the template for the next generation of error-prone PCR; otherwise at the next generation another colony was picked from the same plate. In the unselected populations a single mutant was picked and used as the template for the next generation of error-prone PCR.

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