|Source||Arthrobacter oxydans 25K|
|Substrate specificity||The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.
|Assayed on||DNA pMHaeIII/DriI is a linearized plasmid pMHaeIII. pMHaeIII carries a gene of DNA-methyltransferase M.HaeIII, which methylates sites 5`-GGCC-3` producing 5`-GG(5mC)C-3`/3`-C(5mC)GG-5`.|
|Unit definition||One unit is defined as the amount of enzyme|
required to hydrolyze
in 1 μg of linearized plasmid pMHaeIII/DriI in 1 hour at 60°C in a total reaction volume of 50 μl. As a result a linearized plasmid pMHaeIII/DriI disappears (see run 4 in the figure).
|AoxI activity assay on DNA pMHaeIII/DriI |
1 – Control DNA pMHaeIII/DriI,
2 – 0.5 μl Aox I
3 – 1 μl Aox I
4 – 2 μl Aox I
5 – 1 Kb SE DNA-markers
Products were separated in 1% agarose gel in Buffer TAE.
|Optimal SE-buffer||SE-buffer AoxI (10 mM Tris-HCl (pH 8.5 ??? 25°C); 3 mM MgCl2, 1 mM DTT)|
|Enzyme activity (%)|
|75 - 100||25 - 50||10 - 25||25 - 50||75 - 100||100|
|Reaction buffer||SE-buffer AoxI|
|Storage conditions||10 mM Tris-HCl (pH 7.4); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethan. Store at -20°C.|
|Non-specific hydrolisis||No detectable degradation of 1μg of Lambda DNA was observed after incubation with 1 units of enzyme for 16 hours at 60°C in a total reaction volume of 50 μl.
|Reagents Supplied with Enzyme||
10 X SE-buffer AoxI|
|Methylation sensitivity||The enzyme cleaves C5-methylated DNA and doesn't cut unmodified DNA.|
|Inactivation 20 minutes||No|
|MSDS:||Download MSDS as PDF|
|References:||1. Chernukhin V.A., Belichenko O.A., Tarasova M.V., Gonchar D.A., Akishev A.G., Dedkov V.S., Mikhnenkova N.A., Degtyarev S.Kh. Bacterial strain Arthrobacter oxydans - producer of AoxI site specific endonuclease. // Russian Federation patent RU 2399663 C1 (2009). |