商品编号


EG-1006



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


DNA Modifying Enzymes



Size

4,000 Units

商品信息



Description


Fpg (also known as formamidopyrimidine [fapy]-DNA glycosylase, Mut M, FAPY DNA Glycosylase, and 8-oxoguanine DNA glycosylase) has both N-glycosylase and AP-lyase activities. The N-glycosylase activity releases damaged purines from double stranded DNA, generating an apurinic (AP site). The AP-lyase activity cleaves both 3? and 5? to the AP site thereby removing the AP site and producing a 1 base gap in the DNA and 3′ and 5′ phosphate termini. Bases recognized and removed by Fpg include 7, 8-dihydro-8-oxoguanine (8-oxoguanine), 8-oxoadenine, fapy-guanine, methy-fapy-guanine, fapy-adenine, aflatoxin B1-fapy-guanine, 5-hydroxy-cytosine and 5-hydroxy-uracil (1,2).




Applications



DNA Nicking

DNA Repair

Nick Translation





Source

An
E.coli
strain that carries the cloned
fpg
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 8-oxoguanine base paired with a cytosine in a total reaction volume of 10 μl in 1 hour at 37°C



Components



E. coli fpg:
8,000 U/ml in 20 mM Tris-HCl, pH 8.0 @ 25°C, 50 mM NaCl, 0.5 mM EDTA, 200 μg/ml BSA, 50% Glycerol

10X Reaction Buffer
: 100 mM Bis-Tris-Propane, 100 mM MgCl
2
, 10 mM DTT, pH 7.0 @ 25°C





Quality Control

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



Storage Condition


-20 °C




References



Tchou, J. et al. (1994) Substrate specificity of Fpg protein. J. Biol. Chem., 269, 15318-15324.

Hatahet, Z. et al. (1994) New substrates for old enzymes. J. Biol. Chem., 269, 18814-18820.

Boiteux, S., O’Connor, T. and Laval, J. (1987) Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBO J., 5, 3177-3183.

Singh, N., McCoy, M., Tice, R. and Schneider, L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res., 175, 184-191.

Collins, A., Duthie, S. and Dobson, V. (1993) Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis, 14, 1733-1735.

Collins, A., Dusinska, M., Gedik, C. and Stetina, R. (1996) Oxidative damage to DNA: do we have a reliable bio
Marker
?. Environ. Health Perspect., 104, 465-469.

Pflaum, M., Will, O., Mahler, H.-C. and Epe, B. (1998) DNA oxidation products determined with repair endonucleases in mammalian cells: types, basal levels and influence of cell proliferation. Free Rad. Res., 29, 585-594.

Hartwig, A., Dally, H. and Schlepegrell, R. (1996) Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding. Toxicol. Lett., 88, 85-90.

Czene, S. and Harms-Ringdahl, M. (1995) Detection of single strand breaks and formamidopyrimidine-DNA glycosylase-sensitive sites in DNA of cultured human fibroblasts. Mutat. Res., 336, 235-242.