Sfr274 I

Enzyme nameSfr274 I
PrototypeXhoI
Known analogsBssHI, PaeR7I, SlaI, StrI, TliI, XhoI
SKUSE-E125
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'… C▼TCGAG …3'
3'… GAGCT▲C …5'
SourceStreptomyces fradiae 274
Optimal bufferSE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature50 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10075505075100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (Hind III-digest) in 1 hour at 50°C in a total reaction volume of 50 μl.
Assayed onLambda DNA (Hind III-digest)
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 20-fold overdigestion with enzyme 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 40 u.a. of enzyme for 16 hours at 50°C.
Methylation sensitivityBlocked by CTCG m AG methylation. Not blocked by CT m CGAG methylation.
Reagents supplied with enzyme10 X SE-buffer B.
NotesIncubation at 37°C results in 70% activity
References
  3. L.I. Puchkova, G.N. Krivopalova, I.S. Andreeva, A.V. Selina, G.D. Serov, N.I. Rechkunova, S.Kh. Degtyarev Streptomyces fradiae - a strain producer of restriction endonuclease Sfr274I. // Izvestia SB SA USSR, Biological sciences series, Vol.1, 32-34 (1990). (In Russian)
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