| Recognition site | (5mC)G(5mC)↑NG(5mC)G
G(5mC)GN↓(5mC)G(5mC) |
| Source | Glacial ice bacterium GL24 |
| 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. |
| Optimal 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.) |
| Enzyme % activity: | | B | G | O | W | Y | | 75 - 100 | 75 - 100 | 75 - 100 | 75 - 100 | 75 - 100 |
|
| Optimal temperature | 37oC |
| Assayed on | Double-stranded oligonucleotide
5’ GGTATTC G (5mC) G (5mC)TGTACTTTCCC 3’
3' CCATAAG(5mC) G (5mC) G ACATGAAAGGG 5' |
| 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. |
| Non-specific hydrolisis | 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.
|
| Methylation sensitivity | The enzyme cleaves only methylated DNA. |
| Inactivation 20 minutes under | 65oC |
| MSDS: | Download MSDS as PDF |
| References: |  Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007
V.A. Chernukhin, E.V. Chmuzh, Yu.E. Tomilova, T.N. Nayakshina, D.A. Gonchar, V.S. Dedkov, S.Kh. Degtyarev A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/3’-(5mC)GN^(5mC)G. // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 2, pp 13-17, 2007
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