Endonuclease V is a repair enzyme found in E. coli that recognizes deoxyinosine, a deamination product of deoxyadenosine in DNA. Endonuclease V, often called deoxyinosine 3´ endonuclease, recognizes DNA containing deoxyinosines (paired or not) on double-stranded DNA, single-stranded DNA with deoxyinosines and to a lesser degree, DNA containing abasic sites (ap) or urea, base mismatches, insertion/deletion mismatches, hairpin or unpaired loops, flaps and pseudo-Y structures. It is believed that Endonuclease V needs another protein to repair the DNA, as it does not remove the deoxyinosine or the damaged bases (1,2,3).
Endonuclease V cleaves the second phosphodiester bonds 3´ to the mismatch of deoxyinosine (2), leaving a nick with 3´-hydroxyl and 5´-phosphate (4).
1X NEBuffer™ 4
Incubate at 37°C
1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)
10 mM Tris-HCl
250 mM NaCl
0.1 mM EDTA
50% Glycerol
0.15% Triton® X-100
1 mM DTT
200 µg/ml BSA
pH 8 @ 25°C
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