PspPP I

Enzyme namePspPP I
PrototypePpuMI
Known analogsPpuMI, PpuXI, Psp5II
SKUSE-E255
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'… RG▼GWCCY …3'
3'… YCCWG▲GR …5'
SourcePseudomonas species PP
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
101000100100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA/HindIII in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onLambda DNA/HindIII
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 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 70% of the DNA fragments can be ligated and more than 80% 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 overlapping Dcm methylation(C m CWGG) RGGW CCT GG.
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesTo obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml. Do not use BSA for long incubation.
References
  1. Unpublished data Kileva, E.V., Abdurashitov, M.A., Shevchenko, A.V., Dedkov, V.S., Degtyarev, S.Kh Unpublished observations (1996).
// Код для некролога