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. 2008;3(3):357-62.
doi: 10.1038/nprot.2007.538.

A high-throughput fluorescence-based glycosyltransferase screen and its application in directed evolution

Gavin J Williams  1 , Jon S Thorson
Affiliations

A high-throughput fluorescence-based glycosyltransferase screen and its application in directed evolution

Gavin J Williams et al. Nat Protoc. 2008.

Abstract

This protocol details the application of a high-throughput fluorescence-based screen, in conjunction with error-prone PCR/saturation mutagenesis, for altering the proficiency and/or promiscuity of a secondary metabolite glycosyltransferase (GT) via directed evolution. Given the structural and mechanistic similarities among secondary metabolite-associated GTs, this approach may provide a template for engineering other members of the GT-B superfamily.

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