Description
Recognition site and hydrolysis position:
CCWGG | ↑CCWGG |
GGWCC | GGWCC↓ |
Source: Pseudomonas species 6
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dcm-) in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dcm-)
Optimal SE-buffer: SE-buffer B
Enzyme activity (%):
B | G | O | W | Y | ROSE |
100 | 50 | 10 | 25 | 75 | 10 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; and 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 4 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dcm methylation(CmCWGG): CCWGG.
Notes:
References:
Dedkov, V.S., Tomilova, J.E., Kileva, E.V., Degtyarev, S.K. Unpublished observations. (2002)
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 // Translated from BIOTEHNOLOGIA (Russia) No. 3, pp. 19-24, 2004