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Well-versed investigators describe in step-by-step detail a wide range of DNA repair activities, from single act-alone repair proteins to complex repair systems. These practical protocols not only detail the various repair activities found in cells, but also demonstrate the use of DNA repair proteins and systems as reagents in molecular biology and biotechnology. The techniques described here include mutation and polymorphism detection, which are useful in the search for disease genes and drug response genes, as well as for breeding and trait selection in animals and plants. Compact and highly practical, DNA Repair Protocols: Prokaryotic Systems provides expert guidance to both the DNA repair researcher studying the fundamental aspects of DNA repair and the applied researcher in human genetics and biotechnology.
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Methods in Molecular Biology TM VOLUME 152 DNA Repair Protocols Prokaryotic Systems Edited by Pat Vaughan HUMANA PRESS Repair of A/G and A/8-oxoG Mismatches 3 1 Repair of A/G and A/8-oxoG Mismatches by MutY Adenine DNA Glycosylase A-Lien Lu 1. Introduction Cellular and organism aging have been correlated with accumulated DNA damage (1,2). 8-oxo-7,8-dihydrodeoxyguanine (8-oxoG or GO) is one of the most stable products of oxidative DNA damage. The formation of GO in DNA, if not repaired, can lead to misincorporation of A opposite to the GO lesion and result in G:C to T:A transversions (3–6). In Escherichia coli, a family of enzymes, MutY, MutM, and MutT, is involved in defending against the mutagenic effects of GO lesions (7–9). The E. coli MutY is an adenine glycosylase active on DNA containing A/GO, A/G, and A/C mismatches (7,10–15) and also has a weak guanine glycosylase activity on G/GO-containing DNA (15a,15b). MutY removes misincorporated adenines paired with GO lesions and re