title: Redesigning the single-chain variant of the restriction endonuclease PvuII by circular permutation.
abstract: 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.
authors.affiliation: Institute of Biochemistry, Justus-Liebig University, Heinrich-Buff-Ring 58, D-35392 Giessen, Germany. benno.schierling@chemie.bio.uni-giessen.de
authors.forename: Benno
authors.initials: B
authors.lastname: Schierling
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wende
authors.forename: Alfred
authors.initials: A
authors.lastname: Pingoud
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D053837:DNA Cleavage; D004262:DNA Restriction Enzymes / Q000737:chemistry* / Q000235:genetics; D007700:Kinetics; D009841:Oligonucleotides / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011485:Protein Binding; D015202:Protein Engineering; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D009841:Oligonucleotides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004262:DNA Restriction Enzymes