CONTENTS:
Catalog #/ Description/ Qty
4020-100-01 Beta-polymerase 100 μl
4020-050-02 Beta-pol Control DNA 10 μl
3900-200-08 10X REC Buffer 8 1 ml4020-050-04 Aphidicolin 10 μl
4019-1 REC™ water 1 ml
4018-250 5X REC™ Loading buffer 250 μl
Human DNA Polymerase β is constitutively expressed in cells and functions by filling in gaps in DNA that are formed following base excision repair. The activity of DNA Polymerase β isnot affected by aphidicolin, an inhibitor of DNA polymerasesα, δ, and ε.
SOURCE:
Purified from E. coli containing a recombinant plasmid harboring the human DNA polymerase β gene.
UNIT DEFINITION:
One Unit is the amount of enzyme required to catalyze the incorporation of 1 nmole of dNTP into an acid-insoluble form in 1 hour at 37°C.
SPECIFICITY:
The enzyme can fill small gaps (up to 6 nucleotides) and nicks in DNA, catalyze DNA synthesis after base excision repair, and release 5'-terminal deoxyribose phosphate residues from incised AP sites.
ASSAY CONDITIONS:
1X REC Buffer 8 (50 mM Tris-Cl (pH 8.8), 10 mM MgCl2, 10 mM KCl, 1.0 mM DTT, 1%glycerol) 50 μM dCTP, 50 μM dGTP, 50 μM dATP, 50 μM α-32P-dTTP, and 100 μg/ml ofActivated DNA (Catalog # 4667-50-06 see page 128) in a reaction volume of 100 μl are incubated for 5 min at 37°C.
STORAGE BUFFER:
20 mM Tris-Cl (pH 7.8), 1.0 mM DTT, 1 mM EDTA, 100 mM NaCl, and 50% (v/v) glycerol.
STORAGE:
Store at -20°C in a manual defrost freezer.
REFERENCES:
1. Matsumoto, Y. and K. Kim. 1995. Excision of deoxyribose phosphate residues by DNA polymerase β during DNA repair. Science 269:699-702.
2. Kunkel, T.A. and P.S. Alexander. 1986. The base substitution fidelity of eukaryotic DNA polymerases. J Biol Chem 261:160-166.
3. Jenkins, T.M., J.K. Saxena, A. Kumar, S.H. Wilson, and E.J. Ackerman. 1992. DNA polymerase β and DNA synthesis in Xenopus oocytes and in a nuclear extract.Science 258:475-478.
4. Vens C, E. Dahmen-Mooren, M. Verwijs-Janssen, W. Blyweert, L. Graversen, H. Bartelink, and A.C. Begg. 2002. The role of DNA polymerase beta in determining sensitivity to ionizing radiation in human tumor cells. Nucleic Acids Res.30:2995-3004.
5. Bergoglio V, M.J. Pillaire, M. Lacroix-Triki, B. Raynaud-Messina, Y. Canitrot, A. Bieth, M. Gares, M. Wright, G. Delsol, L.A. Loeb, C. Cazaux, and J.S. Hoffmann. 2002. Deregulated DNA polymerase beta induces chromosome instability and tumorigenesis. Cancer Res. 62:3511-4.
6. Kedar P.S., S.J. Kim, A. Robertson, E. Hou, R. Prasad, J.K. Horton, and S.H. Wilson. 2002. Direct interaction between mammalian DNA polymerase beta and proliferating cell nuclear antigen. J Biol Chem. 277:31115-23. |
