Fsp4H I

Enzyme nameFsp4H I
PrototypeFnu4HI
Known analogsFnu4HI, ItaI, SatI
SKUSE-E095
Turbo versionAvailable
High-concentration versionNot available
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… GC▼NGC …3'
3'… CGN▲CG …5'
SourceAn E.coli strain that carries the cloned Fsp4H I gene from Flavobacterium species 4H
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
50751025100100
Unit definitionOne 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 onLambda DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 200ug/ml BSA; 1mM DTT; 50% glycerol; Store at -20°C.
LigationAfter 5-fold overdigestion with enzyme about 5% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by methylation: 5`-G(5mC)NGC-3`/ 3`-CGN(5mC)G-5`.
Reagents supplied with enzyme10 X SE-buffer Y
References
  3. Unpublished data Dedkov, V.S., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.K Unpublished observations (1995).
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