Directed evolution of novel polymerases.
DNA and RNA polymerases evolved to function in specific environments with specific substrates to propagate genetic information in all living organisms. The commercial availability of these polymerases has revolutionized the biotechnology industry, but for many applications native polymerases are limited by their stability or substrate recognition. Thus, there is great interest in the directed evolution of DNA and RNA polymerases to generate enzymes with novel, desired properties, such as thermal stability, resistance to inhibitors, and altered substrate specificity. Several screening and selection approaches have been developed, both in vivo and in vitro, and have been used to evolve polymerases with a variety of important activities. Both the techniques and the evolved polymerases are reviewed here, along with a comparison of the in vivo and in vitro approaches.
Holmberg
Rebecca C
RC
The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037-1000, USA.
Henry
Allison A
AA
Romesberg
Floyd E
FE
D000818:Animals; D001709:Biotechnology / Q000379:methods; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000235:genetics; D019020:Directed Molecular Evolution / Q000379:methods; D006801:Humans; D013379:Substrate Specificity / Q000235:genetics
D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase