PalA I

Enzyme namePalA I
PrototypeAscI
Known analogsAscI, AsoI, SgsI
SKUSE-E483
Turbo versionNot available
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'… GG▼CGCGCC …3'
3'… CCGCGC▲GG …5'
SourceAn E. coli strain, that carries the cloned gene from Pseudomanas alcaligenes BS17
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
2510101010040
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 KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and 95% of these can be 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 CG methylation.
Reagents supplied with enzyme10 X SE-buffer Y
References
  1. Unpublished data Dedkov, V.S., Belichenko, O.A., Gonchar, D.A., Akishev, A.G., Yamkovaya, T.V., Mezentseva, N.V., Tomilova, J.E., Abdurashitov, M.A., Degtyarev, S.K Unpublished observations(2005)
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