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Construction of Protein Switches by Domain Insertion and Directed Evolution

  • Lucas F. Ribeiro
  • Tiana D. Warren
  • Marc Ostermeier
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1596)

Abstract

A protein switch is a protein that changes between inactive (“off”) and active (“on”) states in response to a biomolecule or physical signal. These switches can be created by fusing two domains in such a way that the activity of the output domain is regulated by the input domain’s recognition of an input signal (such as the binding of a molecule, recognition of light). Here, we describe several methods for randomly fusing two domains to create domain insertion libraries from which protein switches can be identified by selections and/or screens.

Key words

Protein switch Domain insertion Circular permutation Directed evolution 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Lucas F. Ribeiro
    • 1
  • Tiana D. Warren
    • 1
  • Marc Ostermeier
    • 1
  1. 1.Department of Chemical and Biomolecular EngineeringJohns Hopkins UniversityBaltimoreUSA

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