Yeast Genetics

Yeast Genetics: Methods and Protocols is a collection of methods to best study and manipulate Saccharomyces cerevisiae, a truly genetic powerhouse. The simple nature of a single cell eukaryotic organism, the relative ease of manipulating its genome and the ability to interchangeably exist in both haploid and diploid states have always made it an attractive model organism. Genes can be deleted, mutated, engineered and tagged at will. Saccharomyces cerevisiae has played a major role in the elucidation of multiple conserved cellular processes including MAP kinase signaling, splicing, transcription and many others. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Yeast Genetics: Methods and Protocols will provide a balanced blend of classic and more modern genetic methods relevant to a wide range of research areas and should be widely used as a reference in yeast labs.

During the past few decades we have witnessed an era of remarkable growth in the field of molecular biology. In 1950 very little was known of the chemical constitution of biological systems, the manner in which information was trans mitted from one organism to another, or the extent to which the chemical basis of life is unified. The picture today is dramatically different. We have an almost bewildering variety of information detailing many different aspects of life at the molecular level. There great advances have brought with them some breath-taking insights into the molecular mechanisms used by nature for rep licating, distributing and modifying biological information. We have learned a great deal about the chemical and physical nature of the macromolecular nucleic acids and proteins, and the manner in which carbohydrates, lipids and smaller molecules work together to provide the molecular setting of living sys tems. It might be said that these few decades have replaced a near vacuum of information with a very large surplus. It is in the context of this flood of information that this series of monographs on molecular biology has been organized. The idea is to bring together in one place, between the covers of one book, a concise assessment of the state of the subject in a well-defined field. This will enable the reader to get a sense of historical perspectiv(}-what is known about the field today-and a description of the frontiers of research where our knowledge is increasing steadily.

Methods in Yeast Genetics is a course that has been offered annually at Cold Spring Harbor Laboratory for the last 45 years. This is an updated edition of the course manual, which provides a set of teaching experiments, along with protocols and recipes for the standard techniques and reagents used in the study of yeast biology. Since the last edition of the manual was published (2005), revolutionary advances in genomics, proteomics, and imaging technologies have had a significant impact on the field. The 11 experiments included in this manual provide a foundation of methods for any modern-day yeast lab. These methods emphasize combinations of classical and modern genetic approaches, including isolation and characterization of mutants, two-hybrid analysis, tetrad analysis, complementation, and recombination. Also covered are molecular genetic techniques for genome engineering. Additional experiments introduce fundamental techniques in yeast genomics, including both performance and interpretation of Synthetic Genetic Array analysis, multiplexed whole genome and barcode sequencing, and comparative genomic hybridization to DNA arrays. Comparative genomics is introduced using different yeast strains to study natural variation, evolution, and quantitative traits. This manual covers the full repertoire of genetic approaches needed to dissect complex biological problems in the yeast Saccharomyces cerevisiae.

"Methods in Yeast Genetics" is a course that has been offered annually at Cold Spring Harbor for the last 30 years. This provides a set of teaching experiments along with the protocols and recipes for the standard techniques and reagents used in the study of yeast biology.

This fully updated edition of the bestselling three-part Methods in Enzymology series, Guide to Yeast Genetics and Molecular Cell Biology is specifically designed to meet the needs of graduate students, postdoctoral students, and researchers by providing all the up-to-date methods necessary to study genes in yeast. Procedures are included that enable newcomers to set up a yeast laboratory and to master basic manipulations. This volume serves as an essential reference for any beginning or experienced researcher in the field. Provides up-to-date methods necessary to study genes in yeast. Includes proceedures that enable newcomers to set up a yeast laboratory and to master basic manipulations. This volume serves as an essential reference for any beginning or experienced researcher in the field.

Guide to Yeast Genetics and Molecular Biology presents, for the first time, a comprehensive compilation of the protocols and procedures that have made Saccharomyces cerevisiae such a facile system for all researchers in molecular and cell biology. Whether you are an established yeast biologist or a newcomer to the field, this volume contains all the up-to-date methods you will need to study "Your Favorite Gene" in yeast. Key Features * Basic Methods in Yeast Genetics * Physical and genetic mapping * Making and recovering mutants * Cloning and Recombinant DNA Methods * High-efficiency transformation * Preparation of yeast artificial chromosome vectors * Basic Methods of Cell Biology * Immunomicroscopy * Protein targeting assays * Biochemistry of Gene Expression * Vectors for regulated expression * Isolation of labeled and unlabeled DNA, RNA, and protein

The book covers comprehensively all current experimental procedures used in the research of the genetics and molecular biology of the yeast Saccharomyces cerevisiae. Featuring detailed protocols and practical tips, it guarantees easy access to a wide range of specialized topics within thisrapidly advancing field. Internationally-recognized experts present all methods currently in use, discussing topics such as DNA isolulation, cloning and expression vectors, cosmid cloning, construction and use of cDNA libraries, plasmid shuffling and mutant isolulation. Chapters on Ty insertionalmutagenesis, high efficiency transformation, cell-free translation of mRNAs, Ty virus-like particles, and applications to industrial strains of yeast are also included. Researchers in the fields of molecular biology, genetics, and biochemistry working with this yeast, as well as professionals of thebiotechnology industry will refer to this practical reference frequently.

The manual consists of two main sections. The first includes the essential, sometimes laborious, procedures for handling yeasts, for inducing mating and isolation of hybrids, for inducing sporulation and isolation of single-spore clones, with some details of tetrad analysis, and including techniques and ancillary equipment for use of the micromanipulator. There are also proce dures for induction of mutants by physical and chemical agents, and for iso lation of particular types of mutants, such as to temperature sensitivity, for increased frequency of mutations, for mutations in the mitochondrial genome, both to the petite colonie form and to resistance to antibiotics, for mutations in that part of the yeast genome controlling the glycolytic cycle, and numerous others. Mapping of mutations is discussed briefly, though this aspect of yeast genetics is probably one which should not be undertaken until the investigator has gained a certain amount of experience in the field. How ever, as is pointed out in the pertinent part of the manual, the task of mapping has been tremendouslysimplified by the availability from the Yeast Genetics Stock Center at the University of California at Berkeley of a set of auxo trophic strains designed to permit mapping of most unknown genes with a minimum number of crosses and tetrad analyses. The first section concludes with the description of methods for hybridization of yeasts by protoplast fusion, which has been described as the poor man's system for genetic engineering.