Vsp I

Enzyme nameVsp I
PrototypeVspI
Known analogsAseI, PshBI
SKUSE-E139
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'… AT▼TAAT …3'
3'… TAAT▲TA …5'
SourceAn E.coli strain, that carries the cloned gene VspI from Vibrio species 343
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
010501002550
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 7.6); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme 70% of the DNA fragments can be ligated.Of these, 90% can be recut. In the presence of 10% PEG
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 ATTA m AT methylation.
Reagents supplied with enzyme10 X SE-buffer W
References
  2. S.Kh. Degtyarev, E.A. Prikhodko, N.I. Rechkunova, G.G. Prikhodko, V.N. Krasnykh Biochemical characterization of Vsp I methyltransferase. // Gene, 157, 65-66 (1995).
  3. S.Kh. Degtyarev, E.A. Prikhodko, N.I. Rechkunova, Y.A. Gorbunov Interaction of Vsp I and Tru9 I restriction endonuclease with synthetic oligonucleotides. // Biochimica et Biophysica Acta, 1172, 89-94 (1993).
  4. S.Kh. Degtyarev, E.A. Prikhodko, G.G. Prikhodko, Y.A. Gorbunov Vsp I methylase belongs to m6A-gamma class of adenine methylases. // Nucleic Acids Research, Vol. 21, No 8., 2015 (1993).
  5. N.I. Rechkunova, S.Kh. Degtyarev, et al. Preparation method of restriction endonuclease Vsp I // Soviet Union patent SU1641024 (1990). (In Russian)
  6. V.E. Repin, S.Kh. Degtyarev, N.I. Rechkunova, E.V. Zhuk Preparation method of Vibrio species biomass, that enriched by restriction endonuclease Vsp I. // Soviet Union patent SU1559704 (1989). (In Russian)
  7. S.K. Degtyarev, V.E. Repin, N.I. Rechkunova, V.E. Chizhikov, E.G. Malygin, V.V. Mikhailov, V.A. Rasskazov Substrate specificity detrmination of restriction endonuclease VspI. // Bioorg. chem (Moscow), Vol.13, No 3, 420-421 (1987). (In Russian)
// Код для некролога