| Recognition site | ↑CCWGG
GGWCC↓ |
| Source | Acinetobacter johnsonii R2 |
| Assayed on | Lambda DNA |
| 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. |
| Optimal SE-buffer | Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 25 - 50 | 10 - 25 | 10 - 25 | 25 - 50 | 100 | 50 |
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| Optimal temperature | 55oC |
| 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. |
| Ligations | After 3-fold overdigestion with enzyme 95% of the DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | 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.
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| Reagents Supplied with Enzyme |
10 X SE-buffer Y |
| Methylation sensitivity | Not blocked by dcm methylation: CCWGG |
| Inactivation 20 minutes under | 65oC |
| Notes | At 37°C activity is about 10% from maximum.
|
| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: |  V.S. Dedkov, N.A. Mikhnenkova, L.P. Vlasenko, J.E. Tomilova, J.G.Kashirina, D.A. Gonchar, S.Kh. Degtyarev Restriction Endonuclease AjnI from Acinetobacter johnsonii R2, an Isoschizomer of EcoRII, recognizes 5'- CCWGG-3' and cleaves dcm-methylated sites // BIOTEHNOLOGIA (Russia) No. 3, pp. 19-24, 2004
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Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:
To view the fragments length values please point mouse cursor over diagram |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
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