UVDE participates in an alternative excision repair pathway in
which DNA is cut immediately 5’ of CPDs or (6-4) photoproducts.
This contrasts with the nucleotide excision repair pathway in which
damaged bases are removed by DNA incision upstream and downstream
of the damaged site, the gap is filled, and then closed by ligation.
| Source: |
The UVDE-GSTD228 is a fully functional and stable truncated form of the UVDE enzyme from Sz. pombe that is expressed as a fusion protein with glutathione S-transferase (GST). |
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| Specific Activity: |
Lot specific |
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| Unit Definition: |
The unit activity has not been fully characterized, however 250 ng of protein is sufficient to cleave 1 µg of supercoiled UV damaged DNA at 30°C in one hour. |
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| Assay Conditions: |
20 mM HEPES (pH 6.5), 10 mM MgCl2 , 1 mM MnCl2 , and 100 mM NaCl. Reactions of 20 µl are performed at 30°C containing 500 ng of UV-irradiated supercoiled plasmid, and 250 ng of UVDE. Conversion of supercoiled plasmid to open circle and linear forms are detected by agarose gel electrophoresis.
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| Features: |
Highly stable UVDE enzyme
Recognizes both CPD and (6-4) dipyrimidine photoproducts
UVDE is free from contaminating DNase and RNase activity
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| Applications: |
CometAssay™/FLARE™
Supercoiled DNA relaxation assay
Gene specific repair assays
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| Storage: |
Freeze in working aliquots at -80°C to avoid repeated freeze-thaws. The enzyme is supplied in buffer containing 50 mM Tris-Cl (pH 6.0), 10 mM glutathione, and 10% glycerol.
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