By Judith L. Campbell, Paul Modrich
ISBN-10: 012182814X
ISBN-13: 9780121828141
This quantity emphasizes the intracellular results of DNA harm, describing strategies for research of checkpoint responses, DNA fix in vivo, replication fork come upon of DNA harm, in addition to organic tools for research of mutation construction and chromosome rearrangements. It additionally describes molecular equipment for research of a couple of genome upkeep actions together with DNA ligases, helicases, and single-strand binding proteins. *Part B of a 2-part series*Addresses DNA upkeep enzymes*Discusses harm signaling*Presents In vivo research of DNA repair*Covers mutation and chromosome rearrangements
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Chem. 275, 14541–14549. Hanna, J. , Kroll, E. , and Spencer, F. A. (2001). Saccharomyces cerevisiae CTF18 and CTF4 are required for sister chromatid cohesion. Mol. Cell. Biol. 21, 3144–3158. , Flick, J. , and Pexton, T. (1994). Multiple mechanisms provide rapid and stringent glucose repression of GAL gene expression in Saccharomyces cerevisiae. Mol. Cell. Biol. 14, 3834–3841. , Sluis, C. , and Dirr, H. (1997). Conformational stability of pGEX‐expressed Schistosoma japonicum glutathione S‐transferase: A detoxification enzyme and fusion‐protein affinity tag.
1–2 g (pM‐nM) ssDNA, dsDNA (must be linked to chip–usually through a biotin moiety) Rapid, direct assay, can be done under wide range of buffer conditions Requires high salt concentrations for equilibrium binding; subject to experimental artifacts; interpretation can be demanding TO 17 DNA Florescence polarization (FP) 106–109 50 pMoles – 5 g (nM‐M) Fluorescently labeled ssDNA, dsDNA Rapid, direct assay, can be done under wide range of buffer conditions Requires high salt concentrations for equilibrium binding; fluorescence of fluorescein is pH dependent; nonspecific light scattering and reagents with high background fluorescence (agarose or native polyacrylamide gels) and visualized by radioisotope detection or fluorescent imaging.
Sluis, C. , and Dirr, H. (1997). Conformational stability of pGEX‐expressed Schistosoma japonicum glutathione S‐transferase: A detoxification enzyme and fusion‐protein affinity tag. Protein Sci. 6, 399–406. , and Burgers, P. M. (2003). Yeast Rad17/Mec3/Ddc1: A sliding clamp for the DNA damage checkpoint. Proc. Natl. Acad. Sci. USA 100, 2249–2254. , and Burgers, P. M. (2004). The PCNA‐RFC families of DNA clamps and clamp loaders. Progr. Nucl. Acids Res. Mol. Biol. 78, 227–260. Mayer, M. , Gygi, S.
DNA repair by Judith L. Campbell, Paul Modrich
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