PspN4 I

Enzyme namePspN4 I
PrototypeNlaIV
Known analogsBspLI, NlaIV
SKUSE-E089
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'… GGN▼NCC …3'
3'… CCN▲NGG …5'
SourcePseudomonas species N4
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
1010102510050
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); 50 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme more than 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by 5`-GGNN(5mC)C-3`/3-C(5mC)NNGG-5` or 5`-GGNN(5mC)C-3`/3-CCNNGG-5`methylation
Reagents supplied with enzyme10 X SE-buffer Y
References
  3. Unpublished data Abdurashitov, M.A., Kileva, E.V., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh Unpublished observations (1995).
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