Yeast Gene Analysis

Focusing on Saccharomyces cerevisiae, the second edition of Yeast Gene Analysis represents a major reworking of the original edition, with many completely new chapters and major revisions to all previous chapters. Originally published shortly after completion of the yeast genome sequence, the new edition covers many of the major genome-wide strategies that have been developed since then such as microarray analysis of transcription, synthetic gene array studies, protein microarrays and chemical genetic approaches. It represents a valuable resource for any research laboratory using budding yeast as their experimental system in which to identify new yeast gene functions. The chapters are written in a readable style with useful background information, technical tips and specific experimental protocols included as appropriate, enabling both the novice and the experienced yeast researcher to adopt new procedures with confidence. New chapters on: Strain construction; genome-wide two-hybrid approaches; use of microarrays for transcript analysis; real-time analysis of chromosome behaviour and FRET; synthetic gene array technology and protein arrays; chemical genomics and yeast prions; RNA gene analysis and mitochondrial gene function analysis; phylogenetic footprinting; discovering human gene function and predicting yeast gene function

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.

This volume provides a collection of protocols for the study of DNA-DNA contact maps, replication profiles, transcription rates, RNA secondary structures, protein-RNA interactions, ribosome profiling and quantitative proteomes and metabolomes. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Yeast Functional Genomics: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Regulation of gene expression, a major determinant of gene activity, occupies a central place in molecular biology. Yeast Genetic Networks: Methods and Protocols covers approaches to the systems biological analysis of small-scale gene networks in yeast. Divided into four convenient sections, this detailed volume discusses the methods used to analyze gene expression quantitatively, presents a collection of mathematical and computational tools to analyze stochasticity, adaptation, sensitivity in signal transmission, and oscillations in gene expression, provides instructional methods on how to utilize the tools of quantitative genetics to identify genes that regulate stochasticity and oscillations in gene expression, and concludes with a section devoted to the analysis of conserved gene expression systems and networks in different fungal species. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Yeast Genetic Networks: Methods and Protocols serves as an ideal and valuable resource for both novices and experienced researchers.

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.

Yeast Metabolic Engineering: Methods and Protocols provides the widely established basic tools used in yeast metabolic engineering, while describing in deeper detail novel and innovative methods that have valuable potential to improve metabolic engineering strategies in industrial biotechnology applications. Beginning with an extensive section on molecular tools and technology for yeast engineering, this detailed volume is not limited to methods for Saccharomyces cerevisiae, but describes tools and protocols for engineering other yeasts of biotechnological interest, such as Pichia pastoris, Hansenula polymorpha and Zygosaccharomyces bailii. Tools and technologies for the investigation and determination of yeast metabolic features are described in detail as well as metabolic models and their application for yeast metabolic engineering, while a chapter describing patenting and regulations with a special glance at yeast biotechnology closes the volume. Written in the highly successful Methods in Molecular Biology series format, most chapters include an introduction to their respective topic, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Yeast Metabolic Engineering: Methods and Protocols aims to familiarize researchers with the current state of these vital and increasingly useful technologies.