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

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