In vitro directed evolution allows biomolecules with new and useful properties to be engineered�mimicking natural evolution on an experimentally accessible time scale by creating large libraries of DNA mutants using PCR and then carrying out a high-throughput assay for variants with improved function. To provide a breakthrough in the complexity of libraries that can be readily searched experimentally, my laboratory is engineering S. cerevisiae so that both the mutagenesis and selection steps of directed evolution can be carried out entirely in vivo, under conditions of sexual reproduction. I will describe chemical complementation, which provides a selection for chemical transformations beyond those natural to the cell using themes and variations on the yeast two-hybrid assay. Furthermore I will present a heritable recombination system that allows mutations to be introduced and then crossed under conditions of sexual reproduction.

Expanding the synthetic capabilities of the cell

243rd National Meeting (2012)

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