Description
Recognition site and hydrolysis position:
| ACATGT | A↑CATGT |
| TGTACA | TGTAC↓A |
Source: An E.coli strain that carries the cloned Pci I gene from Planococcus citreus SE-F45
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
T7 DNA
Optimal SE-buffer: SE-buffer O
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 50 | 75 | 100 | 75 | 50 | 50 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of T7 DNA with 20 u.a. of enzyme for 16 hours at 37°C.Reagents Supplied with Enzyme10 X SE-buffer O
Reagents Supplied with Enzyme:
Methylation sensitivity: Not blocked by ACmATGT methylation.
Blocked by mACATGT methylation.
Notes: The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,13.
References:
Abdurashitov, M.A., Nayakshina, T.N., Lebedeva, N.A., Dedkov, V.S., Degtyarev, S.K. Unpublished observations (1998).
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.2, No 3, pp 39-46, 2006
