BstKTI, a New dam-Sensitive Neoschizomer of Restriction Endonuclease MboI, which is Able to Cleave Hemimethylated Substrate.
// Biotekhnologia (Moscow), No.2, 5-10 (2006). (In Russian)
BstKT I
| Enzyme name | BstKT I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | MboI | ||||||||||||
| Known analogs | BfuCI, Bsp143I, BstMB I, DpnII, Kzo9 I, MboI, NdeII, Sau3AI | ||||||||||||
| SKU | SE-E151 | ||||||||||||
| Turbo version | Not 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'… GAT▼C …3' 3'… C▲TAG …5' | ||||||||||||
| Source | Bacillus stearothermophilus KT | ||||||||||||
| 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 (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA (dam-) | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C. | ||||||||||||
| Ligation | After 5-fold overdigestion with enzyme more than 90% 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 5 u.a. of enzyme for 16 hours at 37°C. | ||||||||||||
| Methylation sensitivity | Blocked by overlapping Dam methylation (G m ATC) GATC. Not blocked by CG methylation. Cut hemimethylated site:5`- GmATC-3` / 5`-GATC-3` | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer W | ||||||||||||
| References |