Restriction Endonuclease AjnI from Acinetobacter johnsonii R2, an Isoschizomer of EcoRII, recognizes 5′- ↓CCWGG-3′ and cleaves dcm-methylated sites.
// Biotekhnologia (Moscow), No.3, 19-24 (2004). (In Russian)
Ajn I
| Enzyme name | Ajn I | ||||||||||||
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| Prototype | EcoRII | ||||||||||||
| Known analogs | EcoRII, Psp6 I, PspGI, BptI, BseBI, Bst2U I, BstNI, BstOI, MvaI | ||||||||||||
| SKU | SE-E473 | ||||||||||||
| 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'… ▼CCWGG …3' 3'… GGWCC▲ …5' |
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| Source | Acinetobacter johnsonii R2 | ||||||||||||
| Optimal buffer | SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) | ||||||||||||
| Optimal temperature | 55 °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 in 1 hour at 55°C in a total reaction volume of 50 μl. | ||||||||||||
| Assayed on | Lambda DNA | ||||||||||||
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C. | ||||||||||||
| Ligation | After 3-fold overdigestion with enzyme 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 6 u.a. of enzyme for 16 hours at 55°C. | ||||||||||||
| Methylation sensitivity | Not blocked by dcm methylation CCWGG | ||||||||||||
| Reagents supplied with enzyme | 10 X SE-buffer Y | ||||||||||||
| Notes | At 37°C activity is about 10% from maximum. | ||||||||||||
| References |