| Recognition site | G(5mC)N↑GC
(5mC)G↓N(5mC)G |
| Substrate specifity | Optimal substrate (100% activity )
5`-G(5mC)NGC-3`/3`-(5mC)GN(5mC)G-5`
BlsI cleaves DNA sequence 5’-GCNGC-3’, if at least two 5-methylcytosines (N isn’t considering) are present in both strands of the recognition site |
| Source | Bacillus simplex 23 |
| 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. |
| Optimal SE-buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) |
| Enzyme % activity: | | B | G | O | W | Y | | 10 - 25 | 10 - 25 | 50 - 75 | 100 | 75 - 100 |
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| Optimal temperature | 30oC |
| Assayed on | Double-stranded oligonucleotide
5’ CGTGGTGAGAATCAATG(5mC)CGCTAAAGTACAAGC 3’
3' GCACCACTCTTAGTTA(5mC)GG(5mC)GATTTCATGTTCG 5' |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg /ml BSA; 50% glycerol; Store at -20°C. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of pFsp4HI1 DNA (BamHI digest) with 8 unit of BlsI for 16 hours at 30°C. The pFsp4HI1 plasmid carries a gene for Fsp4HI DNA-methyltransferase, which modifies DNA forming 5'-G(5mC)NGC-3'/3'-CGN(5mC)G-5'. |
| 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, Yu.E. Tomilova, E.V. Chmuzh, O.O. Sokolova, V.S. Dedkov, S.Kh. Degtyarev Site-specific endonuclease BlsI recognizes DNA sequence 5’-G(5mC)N^GC-3’ and cleaves it producing 3’ sticky ends // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 1, pp 28-33, 2007
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