Vne I

Enzyme nameVne I
PrototypeApaLI
Known analogsAlw44I, ApaLI
SKUSE-E137
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▼TGCAC …3'
3'… CACGT▲G …5'
SourceAn E.coli strain, that carries the cloned gene VneI from Vibrio nereis 18
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10251002525100
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; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 10-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 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer O
References
  1. S.K. Degtyarev, N.I. Rechkunova, N.A. Netesova, V.E. Chizhikov, E.G. Malygin, N.A. Kochkin, V.V. Mikhailov, V.A. Rasskazov Substrate specificity determination of restriction endonuclease VneI. // Bioorg. Chem (Moscow), Vol.13, No 3, 422-423 (1987). (In Russian)
  2. S.Kh. Degtyarev, N.I. Rechkunova, N.A. Netesova, E.G. Malygin, V.V. Mikhailov, V.A. Rasskazov Vibrio nereis - strain producer of restriction endonuclease VneI, that recognize and cleave DNA sequence 5'-GTGCAC-3'. // Soviet Union patent SU 1413954 (1986). (In Russian)
// Код для некролога