Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents C A T A L O G

Endonuclease V, T.maritima*
* Use of this enzyme in certain applications may be covered by patents and may require a license

#EN0141 Supplied with: 10X Reaction Buffer	250 u (5 u/µl) 0.3 ml
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EN0141 MSDS (English) MSDS (English-USA) MSDS (German)

 

Feature
Optimum activity at 65-70°C.

Description
Endonuclease V, T.maritima, is a 3'-endonuclease, which initiates removal of deaminated bases from damaged DNA, including uracil, hypoxanthine and xanthine. Endonuclease V is also active toward abasic and urea sites, base pair mismatches, flap and pseudo Y structures, and small insertions/deletions in DNA molecules. The cleavage site generated by Endonuclease V occurs at the second phosphodiester bond in the 3' direction from the lesion. When an excess of the enzyme is present, the primary nicked products experience a second nicking event on the complementary strand, leading to a double-stranded break. At low concentrations, Endonuclease V first nicks a DNA strand at the lesions located closer to the 5'-end of the DNA molecule. Single-stranded DNA is cleaved with much lower efficiency than double-stranded DNA. Mg²⁺ or Mn²⁺ ions are required for enzyme activity (1, 2, 3).

Source
E.coli cells carrying a cloned nfi gene of Thermotoga maritima.

Molecular Weight
25 kDa monomer.

Applications

• High-throughput methods for mutation research (3, 4).
• Studies in mutagenesis and DNA repair.
• Mismatch cleavage.
• Genotyping.

Quality Control
The absence of other endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests.

Concentration
5 u/µl

Definition of Activity Unit
One unit of the enzyme converts one µg of supercoiled depurinized plasmid DNA into other topological states in 30 min at 65°C.
Enzyme activity is assayed in the following mixture: 25 mM Na-HEPES (pH 7.4), 5 mM MgCl₂, 5 mM DTT, 2% (v/v) glycerol, 2 µg of partially depurinated pBR322 DNA.

Storage Buffer
The enzyme is supplied in: 20 mM Na-HEPES (pH 7.4), 5 mM DTT, 50 mM NaCl, 0.1% (v/v) Triton X-100 and 50% (v/v) glycerol.

10X Reaction Buffer
250 mM Na-HEPES (pH 7.4), 50 mM MgCl₂, 50 mM DTT, 20% (v/v) glycerol.

Inhibition and Inactivation
Inactivated by heating in boiling water bath for 10 min, preferably in the presence of EDTA, or by phenol/chloroform extraction.

Related Products

Taq DNA Polymerase: recombinant native, without BSA native, with BSA dNTP Mixes dNTP/dUTP Mix dUTP dITP dNTP Set Biotin-11-dUTP Water, nuclease-free

References

1. Huang, J., et al., Multiple cleavage activities of endonuclease V from Thermotoga maritima: recognition and strand nicking mechanism, Biochemistry, 40(30), 8738-8748, 2001.
2. Hitchcock, T.M., et al., Cleavage of deoxyoxanosine-containing oligodeoxyribonucleotides by bacterial endonuclease V, Nucleic Acids Res., 32(13), 4071-4080, 2004.
3. Pincas, H., et al., High sensitivity EndoV mutation scanning through real-time ligase proofreading, Nucleic Acids Res, 32(19), 148, 2004.
4. Huang, J., et al., An endonuclease/ligase based mutation scanning method especially suited for analysis of neoplastic tissue, Oncogene, 21(12), 1909-1921, 2002.