|Methylation sensitivity||The enzyme cleaves only C5-methylated DNA.|
Does not cleave DNA sequence G(4mC)G(4mC).
|Origin||Glacial ice bacterium GL29|
|Storage conditions||10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 0,05% Triton X-100, 0.1 mg/ml BSA, 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 pHspAI DNA (DriI digest) with 8 unit of GlaI for 16 hours at 30°C. The pHspAI plasmid carries a gene for HspAI DNA-methyltransferase, which modifies DNA forming 5'-G(5mC)GC-3'/3'-CG(5mC)G-5'.|
No detectable degradation of a double-stranded labeled oligonucleotide was observed after incubation with 8 unit of restriction endonuclease GlaI for 3 hours at 30°C.
|Optimum temperature||30 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 30°C in a total reaction volume of 20 μl.|
|Inactivation 20 minutes||65oC|
|Optimum SE-buffer||SE-buffer GlaI (10 mM Tris-HCl (pH 8,5 at 25°C); 5 mM MgCl2; 10 mM NaCl; 1 mM 2-mercaptoethanol.)|