Creating hybrid genes by homologous recombination

Article type: Research Article

Authors: Peter L. Wang,

Affiliations: Centre for Protein Engineering and Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, England, UK. Tel.: +44 1223 402104; Fax: +44 1223 402140; E-mail: plw@mrc-lmb.cam.ac.uk

Abstract: Recombination of homologous genes is a powerful mechanism for generating sequence diversity, and can be applied to protein analysis and directed evolution. {\it In vitro} recombination methods such as DNA shuffling are very flexible and can give hybrid genes with multiple crossovers; they have been used extensively to evolve proteins with improved and novel properties. {\it In vivo} recombination in both {\it E. coli} and yeast is greatly enhanced by double-strand breaks; for {\it E. coli}, mutant strains are often necessary to obtain high efficiency. Intra- and inter-molecular recombination {\it in vivo} have distinct features; both give hybrids with one or two crossovers, and have been used to study structure-function relationships of many proteins. Recently {\it in vivo} recombination has been used to generate diversity for directed evolution, creating a large phage display antibody library. Recombination methods will become increasingly useful in light of the explosion in genomic sequence data and potential for engineered proteins.

Journal: Disease Markers, vol. 16, no. 1-2, pp. 3-13, 2000