|Methylation sensitivity||The enzyme cleaves only methylated DNA.|
|Origin||Glacial ice bacterium GL24|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 0,05% Triton X-100, 50% glycerol; Store at -20°C.|
|Assayed on||Double-stranded oligonucleotide|
5’ GGTATTC G (5mC) G (5mC)TGTACTTTCCC 3’
3' CCATAAG(5mC) G (5mC) G ACATGAAAGGG 5'
|Non-specific activity||No nonspecific activity was detected after incubation of 1 μg of pFsp4HI3 DNA (Bgl II digest) with 4 unit of GluI for 16 hours at 37°C. The pFsp4HI3 plasmid carries a double-stranded oligonucleotide 5'- GCCGCGGCAGC -3' / 3'-CGGCGCCGTCG -5' and a gene for M.Fsp4HI DNA-methyltransferase, which modifies DNA forming 5'- G(5mC)CG(5mC)GG(5mC)AGC -3'/3'-CGG(5mC)GC(5mC)GT(5mC)G-5'.|
No detectable degradation of a double-stranded labeled oligonucleotide was observed after incubation with 4 unit of restriction endonuclease GluI for 3 hours at 37°C.
|Optimum temperature||37 oC|
|Unit||One unit is defined as the amount of enzyme required to cleave 1 pmol of the double-stranded oligonucleotide of the above indicated structure in 1 hour at 37°C in a total reaction volume of 20 μl.|
|Inactivation 20 minutes||65oC|
|Optimum SE-buffer||SE-buffer GluI (10 mM Tris-HCl (pH 9,0 at 25°C); 7,5 mM MgCl2; 75 mM NaCl; 1 mM 2-mercaptoethanol.)|