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Mte I
| Enzyme name | Mte I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | Mte | ||||||||||||
| SKU | SE-E553 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Recognition site | 5'… G(5mC)G(5mC)▼NG(5mC)G(5mC) …3'
3'… (5mC)G(5mC)GN▲(5mC)G(5mC)G …5'
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| Source | An E.coli strain that carries the cloned Mte I gene from Microbacterium testaceum 17B | ||||||||||||
| Optimal buffer | SE-buffer W | ||||||||||||
| Optimal temperature | 55 °C | ||||||||||||
| Inactivation temperature | не инактивируется | ||||||||||||
| Buffer activity |
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| 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. | ||||||||||||
| Assayed on | pHspAI10/DriI/metM.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 additionally methylated with M.Fsp4HI includes one site 5`-G(5mC)G(5mC)NG(5mC)G(5mC)-3`/3`-(5mC)G(5mC)GN(5mC)G(5mC)G-5`, which is MteI canonical site [1]. 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 [1]. | ||||||||||||
| 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. | ||||||||||||
| Ligation | |||||||||||||
| Nonspecific hydrolysis | No detectable degradation of 1μg of Lambda DNA was observed after incubation with 40 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. | ||||||||||||
| Supplied with enzyme | 10 X SE-buffer W, DNA pHspAI10/DriI additionally methylated with M.Fsp4HI. | ||||||||||||
| Notes | |||||||||||||
| References |
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Mte I
| Enzyme name | Mte I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | Mte | ||||||||||||
| SKU | SE-E553 | ||||||||||||
| Turbo version | Not available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Recognition site | 5'… G(5mC)G(5mC)▼NG(5mC)G(5mC) …3'
3'… (5mC)G(5mC)GN▲(5mC)G(5mC)G …5'
|
||||||||||||
| Source | An E.coli strain that carries the cloned Mte I gene from Microbacterium testaceum 17B | ||||||||||||
| Optimal buffer | SE-buffer W | ||||||||||||
| Optimal temperature | 55 °C | ||||||||||||
| Inactivation temperature | не инактивируется | ||||||||||||
| Buffer activity |
|
||||||||||||
| 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. | ||||||||||||
| Assayed on | pHspAI10/DriI/metM.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 additionally methylated with M.Fsp4HI includes one site 5`-G(5mC)G(5mC)NG(5mC)G(5mC)-3`/3`-(5mC)G(5mC)GN(5mC)G(5mC)G-5`, which is MteI canonical site [1]. 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 [1]. | ||||||||||||
| 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. | ||||||||||||
| Ligation | |||||||||||||
| Nonspecific hydrolysis | No detectable degradation of 1μg of Lambda DNA was observed after incubation with 40 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. | ||||||||||||
| Supplied with enzyme | 10 X SE-buffer W, DNA pHspAI10/DriI additionally methylated with M.Fsp4HI. | ||||||||||||
| Notes | |||||||||||||
| References |
|