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Redesigning the single‐chain variant of the restriction endonuclease PvuII by circular permutation
Author(s) -
Schierling Benno,
Wende Wolfgang,
Pingoud Alfred
Publication year - 2012
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2012.05.012
Subject(s) - restriction enzyme , endonuclease , genetics , cleavage (geology) , dna , biology , permutation (music) , circular dna , gene , computational biology , recognition sequence , sequence (biology) , genome , physics , paleontology , fracture (geology) , acoustics
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.