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SibEnzyme Ltd. :: Display product information: Gla I
Name
Gla I
Catalog numberE493E494
Package, u.a.100500
Concentration, u.a./ml80008000

Recognition siteG(5mC)^G(5mC)
(5mC)G^(5mC)G
Methylation sensitivityThe enzyme cleaves only C5-methylated DNA.
Does not cleave DNA sequence G(4mC)G(4mC).
OriginGlacial ice bacterium GL29
Storage conditions10 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 -20C.
Assayed onDouble-stranded oligonucleotide
5 GGTATTC  G  (5mC)  G  (5mC)TGTACTTTCCC 3
3' CCATAAG(5mC)  G  (5mC)  G  ACATGAAAGGG 5'
Non-specific activityNo nonspecific activity was detected after incubation of 1 μg of pHspAI DNA (DriI digest) with 8 unit of GlaI for 16 hours at 30C. 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 30C.
Optimum temperature30 oC
UnitOne 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 30C in a total reaction volume of 20 μl.
Inactivation 20 minutes65oC
Optimum SE-bufferSE-buffer GlaI (10 mM Tris-HCl (pH 8,5 at 25C); 5 mM MgCl2; 10 mM NaCl; 1 mM 2-mercaptoethanol.)

Enzyme % activity:
BGOWY
75 - 10075 - 10075 - 10075 - 10090 - 100

Note: The enzyme activity depending on number and position of methylated nucleotides in the recognition sequence (see reference below):
recognition
sequence
G(mc)G(mC)
(mC)G(mC)G
G(mC)G(mC)
CG(mC) G
G(mC)G(mC)
(mc)G C G
G(mC)GC
C G(mC)G
G(mC)G(mC)
C G C G
Activity,%10035-455-62-2,5<0,1



References:

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