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Food spoilage --- Food --- Yeast fungi --- Microbiology

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Apple --- Postharvest diseases and injuries --- Botrytis cinerea --- Antagonists --- Yeast fungi --- Biotechnology

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Yeast Protocols intends to offer a selection of well-proven proto­ cols in cell and molecular biology, applicable to yeasts including, but certainly not exclusively aimed at, Saccharomyces cerevisiae. Saccharomyces cerevisiae and its very distant cousin, Schizosacc- romyces pombe, are of course now foremost model eukaryotic organisms, and the focus of wide-ranging experimental studies, especially those using molecular genetic techniques. Many of the lat­ ter, such as DNA sequencing and in vitro mutagenesis, are general DNA techniques, well covered in other volumes of Humana's Meth­ ods in Molecular Biology series, and elsewhere. The inclusion of a number of non-DNA techniques in this book is meant to reflect the resurgent interest in yeast cell biology sparked by the development of gene manipulation methods—for example, cellular localization of cloned gene products using microscopical techniques. The presentation of protocols follows the successful Methods in Molecular Biology series format, with a clear sequence of steps and extensive troubleshooting notes. It is our hope that these protocols will be useful not only to established members of the full-time research community, but also to the less experienced—first degree level and masters students undertaking project work, as well as PhD students starting their experimental programs; I am well aware that these young apprentice scientists are not always able to receive the supervision time that they, and indeed their supervisors, would like.

Yeasts --- Yeast fungi --- Levures (Botanique) --- physiology --- isolation & purification --- cytology --- Biotechnology --- Research --- Methodology --- Biotechnologie --- Yeast --- -Yeast fungi --- -Yeasts --- Fungi --- Basidiomycetes --- Blastomycetes --- Endomycetales --- Nematospora --- Edible fungi --- Leavening agents --- Yeast-free diet --- -Methodology --- Biotechnology. --- Methodology. --- physiology. --- cytology. --- isolation & purification. --- -Biotechnology --- Cytology. --- Isolation & purification. --- Physiology. --- Yeast biotechnology --- Microbial biotechnology --- Yeast industry --- Research&delete& --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Yeast fungi - Research - Methodology --- Yeast - Biotechnology

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Yeast artificial chromosomes (YACs) have their origins in the molecular genetic analysis of the yeast Saccharomyces cerevisiae. The construction of self-maintaining genetic elements from isolated frag­ ments of the yeast genome defined DNA sequences necessary for chro­ mosome function has provided telomeres, centromeres, and autonomous replicating sequences. In 1987 a reversal of the strategy put these short functional DNA sequences to work in cloning vectors, producing "yeast" chromosomes largely composed of foreign DNA. Initially the insert size of clones averaged several hundred kilobasepairs, a remarkable achieve­ ment. Rapid progress with cloning technology has since enabled the construction of YAC libraries with average insert sizes of around 1 Mb, with many clones exceeding that size, and YACs remain the largest capacity microbiological cloning system available. They effectively bridge the size gap between bacterial cloning (plasmids, cosmids, PI, and bacterial artificial chromosomes) and what could be considered mammalian cloning systems (somatic cell hybrids and irradiati- fusion gene transfer hybrids). YACs also brought with them a conceptual revolution in the man­ agement of clone libraries. The large carrying capacity of YACs, with subsequent reduction in the total number required, meant that it was conceivable to store clones individually rather than as pools that require constant re-plating. Each clone in the library has a unique address and, with successive screenings, information accumulates about individual clones.

Chromosomes, Artificial, Yeast --- Genetic Vectors --- Genetic Techniques --- Molecular cloning --- Genetic vectors --- Yeast fungi --- Saccharomyces cerevisiae --- Vecteurs de clonage --- Methodology --- Genetic engineering --- Genetic vectors. --- Saccharomyces cerevisiae. --- Methodology. --- Genetic engineering. --- Saccharomyces --- Yeast genetic engineering --- Microbial genetic engineering --- Cloning vectors --- Expression vectors (Genetics) --- rDNA vectors --- Recombinant DNA vectors --- Vectors, Genetic --- Gene expression --- Recombinant DNA --- Cloning, Molecular --- DNA cloning --- Gene cloning --- Cloning --- Molecular genetics --- Clone cells --- Human genetics. --- Human Genetics. --- Genetics --- Heredity, Human --- Human biology --- Physical anthropology