商品编号


EG-160



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


DNA Modifying Enzymes



Size

1,000 Units

商品信息



Description


Endonuclease IV (Endo IV) recognizes apurinic/apyrimidinic (AP) sites of dsDNA and cleaves the phosphodiester bond 5' to the lesion generating a hydroxyl group at the 3'-terminus. The enzyme can also act as a 3'-diesterase that is able to release 3'-phosphoglycolate or 3'-phosphate from the damaged ends of dsDNA. Endo IV possesses also a 3'=>5' exonuclease activity. Its progression on substrates is sensitive to ionic strength, metal ions, EDTA, and reducing conditions. Substrates with 3'-recessed ends are preferred substrates for the 3'=>5' exonuclease activity.




Applications



Studies of DNA damage and repair

Single cell electrophoresis (comet assay)

Antitumor drug research

DNA structure research

SNP analysis

DNA Detection Using Recombinase Polymerase Amplification (RPA)





Source

An
E. coli
strain which carries the cloned Endonuclease IV (
nfo)
gene



Unit Definition


One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34-mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 ?l in 1 hour at 37°C.

*An AP site is created by treating 10 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C.




Components



Endonuclease IV:
10,000 U/ml in storage buffer containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 1 mM DTT, 0.05% (v/v) Triton X-100, and 50% (v/v) glycerol.

10X Reaction Buffer
: 1000 mM NaCl, 500 mM Tris-HCl, 100 mM MgCl2, 10 mM DTT, pH 7.9 @ 25°C





Quality Control

The absence of other endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.



Storage Condition


-20 °C




References



?J. D. Levin, Homogeneous Escherichia coli endonuclease IV. Characterization of an enzyme that recognizes oxidative damage in DNA. J. Biol. Chem. 263, 8066-8071 (1988).

S. M. Kerins, et al., Characterization of an endonuclease IV 3’-5’ exonuclease activity. J. Biol Chem. 278(5), 3048-3054 (31 January 2003).

B. Demple, L. Harrison, Repair of oxidative damage to DNA: enzymology and
BIOLOG
y. Annu. Rev. Biochem. 63, 915-948 (1994).

J. D. Levin, B. Demple, In vitro detection of endonuclease IV – specific DNA damage formed by bleomycin in vivo. Nucleic Acids Res. 24, 885-889 (1996)

J. N. Patro, et al., Probing the configurations of formamidopyrimidine lesions Fapy.dA and Fapy.dG in DNA using endonuclease IV. Biochemistry. 43, 13397-13403 (2004).

A. R. Collins, The comet assay for DNA damage and repair: principles, applications, and limitations.Molec. Biotechnol. 26, 249-261 (2004).

D.J. Hosfield et al., Structure of the DNA repair enzyme endonuclease IV and its DNA complex: double-nucleotide flipping at abasic sites and three-metal-ion catalysis. Cell. 98, 397-408 (1999).

I. V. Kutyavin et al., A novel endonuclease IV post-PCR genotyping system. Nucleic Acids Res. 29, 1-9 (2006).