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- W2029580070 abstract "The cloning and propagation of large fragments of DNA on yeast artificial chromosomes (YACs) has become a routine and valuable technique in genome analysis. Unfortunately, many YAC clones have been found to undergo rearrangements or deletions during the cloning process. The frequency of transformation-associated alterations and mitotic instability can be reduced in a homologous recombination-deficient yeast host strain such as a rad52 mutant. RAD52 is one member of an epistatic group of genes required for the recombinational repair of double-strand breaks in DNA. rad52 mutants grow more slowly and transform less efficiently than RAD+ strains and are therefore not ideal hosts for YAC library construction. We have investigated the ability of both null and temperature-sensitive alleles of RAD54, another member of the RAD52 epistasis group, to prevent rearrangements of human YAC clones containing tandemly repeated DNA sequences. Our results show that the temperature-sensitive rad54-3 allele blocks mitotic recombination between tandemly repeated DYZ3 satellite sequences and significantly stabilizes a human DYZ5 satellite-containing YAC clone. Yeast carrying the rad54-3 mutation can undergo meiosis, have growth and transformation rates comparable with RAD+ strains, and therefore represent improved YAC cloning hosts." @default.
- W2029580070 created "2016-06-24" @default.
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- W2029580070 date "1997-03-15" @default.
- W2029580070 modified "2023-10-18" @default.
- W2029580070 title "Stabilization of yeast artificial chromosome clones in a rad54-3 recombination-deficient host strain" @default.
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- W2029580070 doi "https://doi.org/10.1093/nar/25.6.1248" @default.
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