title: Construction of Protein Switches by Domain Insertion and Directed Evolution.
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.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St, Baltimore, MD, 21218, USA.
authors.forename: Lucas F
authors.initials: LF
authors.lastname: Ribeiro
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St, Baltimore, MD, 21218, USA.
authors.forename: Tiana D
authors.initials: TD
authors.lastname: Warren
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St, Baltimore, MD, 21218, USA. oster@jhu.edu.
authors.forename: Marc
authors.initials: M
authors.lastname: Ostermeier
mesh_terms: D005821:Genetic Techniques; D000072417:Protein Domains / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011506:Proteins / Q000235:genetics*
chemical_list: D011506:Proteins
keywords: Circular permutation; Directed evolution; Domain insertion; Protein switch