By Daryl S. Henderson
Univ. of Dundee, united kingdom. offers distinctive directions for learning manifold elements of the eukaryotic reaction to genomic damage. Emphasizes tools for interpreting DNA fix methods in mammalian cells. For researchers. Hardcover, plastic-comb binding additionally to be had.
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Extra resources for DNA Repair Protocols: Eukaryotic Systems
When a Transtar-96 multipipeter is not available, use the sterile cartridges as “stamps” to transfer cells. 4. hprt is an X-linked gene encoding a nonessential purine salvage pathway enzyme, hypoxanthine phosphoribosyl transferase (HPRT). HPRT metabolizes TG to a cytotoxic nucleotide. See ref. (8) for a detailed discussion of hprt mutagenesis. 54 Zdzienicka 5. When preparing the master plate, transfer no more than 12–24 clones to a part of the 96-well plate, trypsinize them, and add standard medium before transferring the next 12–24 clones.
Mutants defective in dark repair (uvh1, uvr1, uvr5, and uvr7) and in the photoreactivation of CPDs (uvr2) and (6-4)PPs (uvr3) have been identified and, with the exception of uvr3, mapped, and the genes corresponding to the photolyase mutations have been cloned and sequenced. The genes From: Methods in Molecular Biology, Vol. 113: DNA Repair Protocols: Eukaryotic Systems Edited by: D. S. , Totowa, NJ 31 32 Britt and Jiang corresponding to the dark repair mutations, presumably homologs of the excision repair genes of yeast and mammals, have not yet been isolated.
7. All mutagen-sensitive lines should be recloned to avoid possible contamination with other cells. 8. For further studies, use cells with a significantly increased sensitivity to the given agent. To date, the isolated mutants show a 2- to 10-fold increased sensitivity toward X-rays or UV radiation, and more than 10-fold to MMC. References 1. Zdzienicka, M. Z. and Simons, J. W. I. M. (1987) Mutagen-sensitive cell lines are obtained with a high frequency in V79 Chinese hamster cells. Mutat. Res.