Acc65 I

Enzyme nameAcc65 I
PrototypeKpnI
Known analogsAsp718I, Kpn I
SKUSE-E003
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▼GTACC …3'
3'… CCATG▲G …5'
SourceAcinetobacter calcoaceticus 65
Optimal bufferSE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10257510010100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA(dcm-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onLambda DNA(dcm-)
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 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 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by overlapping Dcm methylation(C m CWGG) GGTA CC WGG.
Reagents supplied with enzyme10 X SE-buffer W
NotesHigh enzyme concentration may result in star activity.
References
  2. V.E. Repin, S.K. Degtyarev, N.I. Rechkunova Acinetobacter calcoaceticus 65 - a strain producer of restriction endonuclease, that recognize and cleave sequence 5′-GGTACC-3′. // Russian Federation patent RU2034922 (1995) (In Russian).
  3. G.G. Prikhodko, N.I. Rechkunova, V.E. Repin, S.Kh. Degtyarev Substrate specificity determination of restriction endonuclease Acc65I. // Sibirian Biological Journal, No 1, 59-60 (1991). (In Russian)
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