Purification and characterization of the in vitro activity of I-Sce I, a novel and highly specific endonuclease encoded by a group I intron

C Monteilhet; A Perrin; A Thierry; L Colleaux; B Dujon

doi:10.1093/nar/18.6.1407

Purification and characterization of the in vitro activity of I-Sce I, a novel and highly specific endonuclease encoded by a group I intron

Nucleic Acids Res. 1990 Mar 25;18(6):1407-13. doi: 10.1093/nar/18.6.1407.

Authors

C Monteilhet¹, A Perrin, A Thierry, L Colleaux, B Dujon

Affiliation

¹ Unité de Génétique moléculaire des Levures, Institut Pasteur, Paris, France.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Chromatography, Gel
Chromatography, Ion Exchange
Deoxyribonucleases, Type II Site-Specific / genetics
Deoxyribonucleases, Type II Site-Specific / isolation & purification*
Deoxyribonucleases, Type II Site-Specific / metabolism
Enzyme Stability
Escherichia coli / enzymology
Escherichia coli / genetics
Introns*
Kinetics
Molecular Sequence Data
Molecular Weight
Plasmids
Saccharomyces cerevisiae / enzymology*
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae Proteins

Substances

Saccharomyces cerevisiae Proteins
SCEI protein, S cerevisiae
Deoxyribonucleases, Type II Site-Specific