Redesigning the single-chain variant of the restriction endonuclease PvuII by circular permutation.

Benno Schierling, Wolfgang Wende, Alfred Pingoud

Institute of Biochemistry, Justus-Liebig University, Heinrich-Buff-Ring 58, D-35392 Giessen, Germany. benno.schierling@chemie.bio.uni-giessen.de

FEBS letters 2012 Jun 12

filter terms:

The restriction endonuclease PvuII has been introduced as a sequence-specific cleavage module in highly-specific nucleases for gene targeting. Here, a structural reorganization of the single-chain variant of PvuII (scPvuII) was performed by circular permutation as a proof-of-concept in order to find out whether the relocated, new termini next to structural elements important for DNA recognition and catalysis could be used for the fusion with other regulatory protein domains. Three circularly permuted variants of scPvuII were obtained that all maintain the specific endonucleolytic activity of scPvuII. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Citation

Benno Schierling, Wolfgang Wende, Alfred Pingoud. Redesigning the single-chain variant of the restriction endonuclease PvuII by circular permutation. FEBS letters. 2012 Jun 12;586(12):1736-41