Description
Recognition site and hydrolysis position:
| VCTCGAGB | VC↑TCGAGB |
| BGAGCTCV | BGAGCT↓CV |
Source: An E.coli strain that carries the cloned PspX I gene from Pseudomonas species A1-1
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (HindIII-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (HindIII-digest)
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):
| B | G | O | W | Y | ROSE |
| 50 | 50 | 25 | 75 | 100 | 25 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM KCl; 0.1 mM EDTA; 7 mM 2 -mercaptoethanol, 200 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.Reagents Supplied with Enzyme
10 X SE-buffer Y, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Gonchar D.A., Abdurashitov M.A., Belichenko O.A., Dedkov V.S., Mezentseva N.V., Tomilova J.E., Degtyarev S.Kh. Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov, No. 1, pp18-23 (2005)
