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Protein trans ‐splicing on an M13 bacteriophage: towards directed evolution of a semisynthetic split intein by phage display by Daniel Garbe; Ilka V. Thiel; Henning D. Mootz is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Protein trans ‐splicing on an M13 bacteriophage: towards directed evolution of a semisynthetic split intein by phage display about?

## Abstract Split inteins link their fused peptide or protein sequences with a peptide bond in an autocatalytic reaction called protein __trans__‐splicing. This reaction is becoming increasingly important for a variety of applications in protein semisynthesis, polypeptide circularisation, construction of biosensors, or segmental isotopic labelling of proteins. However, split inteins exhibit greatly varying solubility, efficiency and tolerance towards the nature of the fused sequences as well as reaction conditions. We envisioned that phage display as an __in vitro__ selection technique would provide a powerful tool for the directed evolution of split inteins with improved properties. As a first step towards this goal, we show that presentation of active split inteins on an M13 bacteriophage is feasible. Two different __C__‐terminal intein fragments of the __Ssp__ DnaB intein, artificially split at amino acid positions 104 and 11, were encoded in a phagemid vector in fusion to a truncated gpIII protein. For efficient production of hybrid phages, the presence of a soluble domain tag at their __N__‐termini was necessary. Immunoblot analysis revealed that the hybrid phages supported pr

Who reads Protein trans ‐splicing on an M13 bacteriophage: towards directed evolution of a semisynthetic split intein by phage display?

Author

Daniel Garbe; Ilka V. Thiel; Henning D. Mootz

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 1075-2617)

Published

2010

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)