Abbreviations:BHQ1 (black hole fluorescence quencher 1), DMDA (N,N-dimethylethylenediamine), 6FAM (6-catboxyfluorescein), HhH (helix–hairpin–helix), hTDG (human TDG), hyTDG (hybrid TDG), 5-mC (5-methylcytosine), MIG (thymine DNA glycosylase from Methanobacterium thermoautotrophicum), MS (mass spectrometry), NCI (negative chemical ionization), TBDMS (t-butyldimethylsilyl), TDG (thymine DNA glycosylase), UDG (uracil-DNA glycosylase)
Construction and characterization of a hybrid human-thermophile mispaired TDG (hyTDG)
Examination of the activity of hyTDG using a gel cleavage assay
Examination of the activity of hyTDG using a real-time fluorescence assay
where Yt is percent cleavage at time t (min), A is the percent cleavage at 60 min, and k is a rate constant (min−1).
Examination of pyrimidines released from oligonucleotides and DNA by hyTDG
- Costello M.
- Pugh T.J.
- Fennell T.J.
- Stewart C.
- Lichtenstein L.
- Meldrim J.C.
- Fostel J.L.
- Friedrich D.C.
- Perrin D.
- Dionne D.
- Kim S.
- Gabriel S.B.
- Lander E.S.
- Fisher S.
- Getz G.
Stable isotope standards
Construction, cloning, and purification of the hyTDG
Characterization of the purified hyTDG by LC–MS/MS analysis
Gel-based cleavage assay
Real-time fluorescence assay
Oligonucleotide cleavage assays monitored by gel and GC–MS/MS
Preparation of calf thymus DNA and analysis of base composition
Analysis of bases released from calf thymus DNA by UDG and hyTDG
- Vizcaíno J.A.
- Côté R.G.
- Csordas A.
- Dianes J.A.
- Fabregat A.
- Foster J.M.
- Griss J.
- Alpi E.
- Birim M.
- Contell J.
- O'Kelly G.
- Schoenegger A.
- Ovelleiro D.
- Pérez-Riverol Y.
- Reisinger F.
- et al.
Conflict of interest
Funding and additional information
- Supporting information
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