The Pse31I restriction endonuclease from the bacterial strain Peribacillus species 31, an isozomere of Eco31I, recognizes the non-palindromic DNA sequence 5’-GGTCTC-3’ (1/5)
// DNA-Recognizing Enzymes, Vol. 2025(1), DOI: 10.26213/3034-4301.2025.4.1.001 (2025). (In Russian)
Bso31 I
| Enzyme name | Bso31 I | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Prototype | Eco31I | ||||||||||||
| Known analogs | BsaI, BspTNI, Eco31I | ||||||||||||
| SKU | SE-E285 | ||||||||||||
| Turbo version | Available | ||||||||||||
| High-concentration version | Not available | ||||||||||||
| Restriction type | Type IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly. | ||||||||||||
| Recognition site | 5'… GGTCTCN▼ …3' 3'… CCAGAG(N)5▲ …5' | ||||||||||||
| Source | Bacillus stearothermophilus 31 | ||||||||||||
| Optimal buffer | SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) | ||||||||||||
| Optimal temperature | 55 °C | ||||||||||||
| Inactivation temperature | 80 °C | ||||||||||||
| Buffer activity |
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| Unit definition | One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 55°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | T7 DNA | ||||||||||||
| 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 DNA fragments can be ligated. Of these 80% can be recut. | ||||||||||||
| Nonspecific hydrolysis | No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 10 u.a. of enzyme for 16 hours at 55°C. | ||||||||||||
| Methylation sensitivity | Blocked by overlapping Dcm-methylation (CmCWGG) GAGA CC WGG Not blocked by methylation GGTCT(5mC) | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer O, BSA | ||||||||||||
| Notes | To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation. | ||||||||||||
| References |