HspA I

Enzyme nameHspA I
PrototypeHhaI
Known analogsHin6I, HinP1I, AspLE I, BstHH I, CfoI, HhaI
SKUSE-E069
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'… G▼CGC …3'
3'… CGC▲G …5'
SourceAn E.coli strain that carries the cloned HspA I gene from Haemophilus species A1
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 temperature80 °C
Buffer activity
BGOWYROSE
50502525100100
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 NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 20-fold overdigestion with enzyme more than 90% 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 CG methylation 5'-G(5mC)GC-3'/3'-CG(5mC)G-5'. Not blocked by methylation 5'-GCG(5mC)-3'/3'-CGCG-5'. Cut hemimethylated site:5'- G(5mC)GC-3'/3'-CGCG-5`
Reagents supplied with enzyme10 X SE-buffer Y
References
  3. Unpublished data Rechkunova, N.I., Prikhod'ko, E.A., Shevchenko, A.V., Degtyarev, S.K Unpublished observations (1994).
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