|Source||Microbacterium testaceum 17B|
|Substrate specificity||The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.
|Assayed on||pHspAI10/DriI+M.Fsp4HI is a plasmid pHspAI10, which is linearized with DriI, and, additionally, modified with Fsp4HI DNA methyltransferase. pHspAI10 carries a gene of HspAI DNA methyltransferase, that modifies the sequence |
5`-GCGC-3`, producing 5`-G(5mC)GC-3`.
M.Fsp4HI modifies the sequence
5`-GCNGC-3`, producing 5`-G(5mC)NGC-3`.
A substrate pHspAI10/DriI+M.Fsp4HI includes one site
which is MteI canonical site . The enzyme activity depends on a number and positions of methylated nucleotides in the recognition sequence. For example, MteI cuts the recognition site with six 5-methylcytosines, but the enzyme activity is reduced for more that one order .
|Unit definition||One unit is defined as the amount of enzyme required to hydrolyze completely 1 μg of linearized plasmid pHspAI10/DriI+M.Fsp4HI in 1 hour at 55°C in a total reaction volume of 50 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 (%)|
|25 - 50||75 - 100||75 - 100||100||50 - 75||100|
|Reaction buffer||SE-buffer W, (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)|
|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 detectable degradation of 1μg of Lambda DNA was observed after incubation with 10 units of enzyme for 16 hours at 55°C in a total reaction volume of 50 μl.
|Methylation sensitivity||The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.|
|Inactivation 20 minutes||No|
|MSDS:||Download MSDS as PDF|
|References:||1. V.A. Chernukhin, E.V. Kileva, V.A. Sokolova., D.A. Gonchar, L.N. Golikova, V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev A new methyl-directed site-specific DNA endonuclease MteI cleaves nine nucleotides sequence 5’-G(5mC)G(5mC)^NG(5mC)GC-3’/3’-CG(5mC)GN^(5mC)G(5mC)G-5’ // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.8, No 1, pp 16-26, 2012