caa a22 4500 445821523 CHVBK 20180317145245.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s00239-010-9415-2 doi (NATIONALLICENCE)springer-10.1007/s00239-010-9415-2 Exploiting Models of Molecular Evolution to Efficiently Direct Protein Engineering [Elektronische Daten] [Megan Cole, Eric Gaucher] Directed evolution and protein engineering approaches used to generate novel or enhanced biomolecular function often use the evolutionary sequence diversity of protein homologs to rationally guide library design. To fully capture this sequence diversity, however, libraries containing millions of variants are often necessary. Screening libraries of this size is often undesirable due to inaccuracies of high-throughput assays, costs, and time constraints. The ability to effectively cull sequence diversity while still generating the functional diversity within a library thus holds considerable value. This is particularly relevant when high-throughput assays are not amenable to select/screen for certain biomolecular properties. Here, we summarize our recent attempts to develop an evolution-guided approach, Reconstructing Evolutionary Adaptive Paths (REAP), for directed evolution and protein engineering that exploits phylogenetic and sequence analyses to identify amino acid substitutions that are likely to alter or enhance function of a protein. To demonstrate the utility of this technique, we highlight our previous work with DNA polymerases in which a REAP-designed small library was used to identify a DNA polymerase capable of accepting non-standard nucleosides. We anticipate that the REAP approach will be used in the future to facilitate the engineering of biopolymers with expanded functions and will thus have a significant impact on the developing field of ‘evolutionary synthetic biology'. Springer Science+Business Media, LLC, 2010 Directed evolution nationallicence Evolutionary models nationallicence Functional divergence nationallicence Protein engineering nationallicence Cole Megan School of Biology, Georgia Institute of Technology, 30332, Atlanta, GA, USA aut Gaucher Eric School of Biology, Georgia Institute of Technology, 30332, Atlanta, GA, USA aut Journal of Molecular Evolution Springer-Verlag 72/2(2011-02-01), 193-203 0022-2844 72:2<193 2011 72 239 https://doi.org/10.1007/s00239-010-9415-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00239-010-9415-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cole Megan School of Biology, Georgia Institute of Technology, 30332, Atlanta, GA, USA aut NATIONALLICENCE 700 1- Gaucher Eric School of Biology, Georgia Institute of Technology, 30332, Atlanta, GA, USA aut NATIONALLICENCE 773 0- Journal of Molecular Evolution Springer-Verlag 72/2(2011-02-01), 193-203 0022-2844 72:2<193 2011 72 239 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer