Expanding the substrate repertoire of a DNA polymerase by directed evolution

M. Fa, A. Radeghieri, A.A. Henry, F.E. Romesberg, J. Am. Chem. Soc. (2004) 126:1748-1754.
pubpic2004faWe identify mutants that efficiently synthesize an unnatural polymer from 2‘-O-methyl ribonucleoside triphosphates using an activity-dependent phage display method. One evolved polymerase is shown to incorporate the modified substrates with an efficiency and fidelity equivalent to that of the wild-type enzyme with natural substrates.