Open AccessPurification and characterization of thein vitroactivity of I-SceI, a novel and highly specific endonuclease encoded by a group I intron
Author(s) -
Claude Monteilhet,
Arnaud Perrin,
Agnès Thierry,
Laurence Colleaux,
Bernard Dujon
Publication year - 1990
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/18.6.1407
Subject(s) - biology , humanities , genetics , microbiology and biotechnology , art
Group I intron encoded proteins represent a novel class of site specific double strand endonucleases. The endonuclease activity of this class of proteins has been first demonstrated in vivo for I-Sce I which is encoded by a mitochondrial intron of Saccharomyces cerevisiae. Assays using crude cell extracts have shown that I-Sce I can be used in vitro as a restriction endonuclease potentially useful for recombinant DNA technology owing to its large recognition sequence (18 nucleotides). We report here the purification and the first detailed analysis of the in vitro activity and properties of I-Sce I.