Directed evolution of proteins in vitro using compartmentalization in emulsions

Eric A. Davidson, Paulina J. Dlugosz, Matthew Levy, Andrew D. Ellington

Research output: Contribution to journalReview article

5 Scopus citations

Abstract

This unit describes a protocol for the directed evolution of proteins utilizing in vitro compartmentalization. This method uses a large number of independent in vitro transcription and translation (IVTT) reactions in water droplets suspended in an oil emulsion to enable selection of proteins that bind a target molecule. Protein variants that bind the target also bind to and allow recovery of the genes that encoded them. This protocol serves as a basis for carrying out selections in emulsions, and can potentially be modified to select for other functionalities, including catalysis. This selection method is advantageous compared to alternative selection protocols due to the ability to screen through very large-size libraries and the ability to express and screen or select for functions that would otherwise be toxic or inaccessible to in vivo selections and screens.

Original languageEnglish (US)
Pages (from-to)24.6.1-24.6.12
JournalCurrent Protocols in Molecular Biology
Issue numberSUPPL. 87
DOIs
Publication statusPublished - Jul 29 2009

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Keywords

  • Directed evolution
  • Emulsion
  • In vitro compartmentalization

ASJC Scopus subject areas

  • Molecular Biology

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