Zra I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned Zra I gene from Zoogloea ramigera 11
Unit definition:
One 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 on:
Lambda DNA
Optimal SE-buffer: SE-buffer B
Enzyme activity (%):

Storage conditions: 10 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.
Ligations:
After 10-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better.
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 B
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: High enzyme concentration may result in star activity.The minimum number of units that resulted in complete digestion of 1 ug of substrate DNA in 16 hours is 0,5.ZraI cleaves linear plasmid DNA at a rate 1,5-2 times higher than supercoiled plasmid DNA.
References:
Dedkov, V.S., Sinichkina, S.A., Popichenko, D.V., Degtyarev, S.K. Biotekhnologia 6 : 3-7 (2001)