MRNA display for the selection and evolution of enzymes from in vitro-translated protein libraries

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Abstract

The mrnadisplay technology enables the in vitroselection and directed evolution of functional proteins from libraries of more than 1012different mutants in a single test tube. the size of these libraries is well beyond the limit of screenin g technologiesand of most in vivoand in vitroselection methods. the mrnadisplay technology has been used to select peptides and proteins that bind to a specific ligand, as well as novel enzymes. this protocol details the procedure to produce mrna-displayed proteins (3 d) and to subject them to a selection and evolution of enzymes for bond-forming reactions (4-10 weeks). this method is demonstrated by the generation of new rnaligase enzymes.

Original languageEnglish (US)
Pages (from-to)540-552
Number of pages13
JournalNature Protocols
Volume6
Issue number4
DOIs
StatePublished - Apr 2011

Bibliographical note

Funding Information:
acknowleDGMents The author thanks J.W. Szostak and members of the Szostak lab for their advice and help. The author is supported in part by the National Aeronautics and Space Administration under Agreement No. NNX09AH70A issued through the NASA Astrobiology Institute/Ames Research Center.

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