Restriction endonuclease BmtI from Bacillus megaterium S2 cleaves DNA in 5′-GCTAG^C-3′.
// Biotekhnologia (Moscow), No.1, P. 11-15 (2003). (In Russian)
Bmt I
| Enzyme name | Bmt I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | NheI | ||||||||||||
| Known analogs | AsuNH I, NheI | ||||||||||||
| SKU | SE-E457 | ||||||||||||
| Turbo version | Available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Restriction type | Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites. | ||||||||||||
| Recognition site | 5'… GCTAG▼C …3' 3'… C▲GATCG …5' |
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| Source | An E.coli strain that carries the cloned Bmt I gene from Bacillus megaterium S2 | ||||||||||||
| Optimal buffer | SE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) | ||||||||||||
| Optimal temperature | 37 °C | ||||||||||||
| Inactivation temperature | 65 °C | ||||||||||||
| Buffer activity |
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| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA (Hind III-digest) | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C. | ||||||||||||
| Ligation | After 20-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | not tested | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer W | ||||||||||||
| Notes | The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,13.BmtI cleaves linear plasmid DNA at a rate 5 times higher than supercoiled plasmid DNA. | ||||||||||||
| References |