Description
Recognition site and hydrolysis position:
GCNGC | GC↑NGC |
CGNCG | CGN↓CG |
Source: An E.coli strain that carries the cloned Fsp4H I gene from Flavobacterium species 4H
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
50 | 75 | 10 | 25 | 100 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 200ug/ml BSA; 1mM DTT; 50% glycerol; Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme about 5% 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 5 u.a. of enzyme for 16 hours at 37°C.Reagents Supplied with Enzyme
10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by methylation: 5`-G(5mC)NGC-3`/ 3`-CGN(5mC)G-5`.
Notes:
References:
Dedkov, V.S., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.K. Unpublished observations (1995).
Chmuzh E.V., Kashirina Yu.G., Tomilova Yu.E., Chernukhin V.A., Okhapkina S.S., Gonchar D. A., Dedkov V.S., Abdurashitov M. A., Degtyarev S. Kh. Molecular Biology, V.41, No 1, pp 43-50 (2007) (In Russian)
Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 4, pp 19-27, 2007
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006