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

Gavin J. Williams, Jon S. Thorson

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

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.

Original languageEnglish
Pages (from-to)357-362
Number of pages6
JournalNature Protocols
Volume3
Issue number3
DOIs
StatePublished - Mar 2008

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS We are grateful to the School of Pharmacy Analytical Instrumentation Center for analytical support. This work was supported in part by National Institutes of Health Grants AI52218 and U19 CA113297. J.S.T is a UW HI Romnes Fellow.

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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