In vitro protein evolution

M. Zaccolo and E. Gherardi, The effect of high-frequency random mutagenesis on in vitro protein evolution a study on TEM-1 beta-lactamase, J. Mol. Biol. 1999, 285, 775-783. 

RATIONAL EVOLUTIONARY DESIGN THE THEORY OF In Vitro PROTEIN EVOLUTION... 

To incorporate the benefits of neutrality into in vitro protein evolution experiments, it is necessary to understand the interactions between neutral networks of different structures and functions. In Section IV.B.l, we describe the structure of neutral networks based on measures developed primarily for RNA secondary structure landscapes. In Section IV.B.2, we focus on mutational transitions between networks. The behavior of evolving populations on neutral networks is described in Section IV.B.3, with the intention of driving future directed evolution experiments. 

The benefit of neutrality has yet to be captured with in vitro protein evolution. Neutral theory predicts the punctuated emergence of novel structure and function, however, with current methods, the required time scale is not feasible. Utilizing neutral evolution to accelerate the discovery of new functional and structural solutions requires a theory that predicts the behavior of mutational pathways between networks. Because the transition from neutral to adaptive evolution requires a multi-mutational switch, increasing the mutation rate decreases the time required for a punctuated change to occur. By limiting the search to... 

II. The Key to In Vitro Protein Evolution Cell-free Translation. 372... 

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