Yeast Protocols : Methods in Cell and Molecular Biology
Yeast Protocols : Methods in Cell and Molecular Biology
edited by Ivor H. Evans.
- 1st edition 1996
- 1 recurso en línea (XIII, 433 páginas)
- Methods in Molecular Biology 53 1940-6029 .
Isolation and Identification of Yeasts from Natural Habitats -- Maintenance and Culture of Yeasts -- Mutagenesis in Yeast -- Rare-Mating and Cytoduction in Saccharomyces cerevisiae -- Protoplast Fusion in Saccharomyces cerevisiae -- Meiotic Analysis -- Ascus Dissection -- Karyotyping of Yeast Strains by Pulsed-Field Gel Electrophoresis -- Chromosomal Localization of Genes Through Pulsed-Field Gel Electrophoresis Techniques -- Isolation, Purification, and Analysis of Nuclear DNA in Yeast Taxonomy -- Isolation of Yeast DNA -- Isolation of Mitochondrial DNA -- Isolation of Yeast Plasma Membranes -- The Extraction and Analysis of Sterols from Yeast -- Peroxisome Isolation -- Transformation of Lithium-Treated Yeast Cells and the Selection of Auxotrophic and Dominant Markers -- Biolistic Transformation of Yeasts -- Genomic Yeast DNA Clone Banks -- Yeast Colony Hybridization -- Determination of Plasmid Copy Number in Yeast -- Determination of Chromosome Ploidy in Yeast -- Chromosome Engineering in Yeast with a Site-Specific Recombination System from a Heterologous Yeast Plasmid -- Insertional Mutagenesis by Ty Elements in Saccharomyces cerevisiae -- Reporter Gene Systems for Assaying Gene Expression in Yeast -- Preparation and Use of Yeast Cell-Free Translation Lysate -- Electrophoretic Analysis of Yeast Proteins -- Preparation of Total RNA -- mRNA Abundance and Half-Life Measurements -- Polysome Analysis -- Induction of Heat Shock Proteins and Thermotolerance -- Rhodamine B Assay for Estimating Activity of Killer Toxins Permeabilizing Cytoplasmic Membranes -- Nystatin-Rhodamine B Assay for Estimating Activity of Killer Toxin from Kluyveromyces lactis -- Application of Killer Toxins in Stepwise Selection of Hybrids and Cybrids Obtained by Induced Protoplast Fusion -- Killer Plaque Technique for Selecting Hybrids and Cybrids Obtained by Induced Protoplast Fusion -- Genotoxicity Testing in Schizosaccharomyces pombe -- Purification and Quantification of Saccharomyces cerevisiae Cytochrome P450 -- Calorimetry of Whole Yeast Cells -- Light Microscopy Methods -- Fluorescence Microscopy Methods -- Immunoelectron Microscopy.
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.
9781592595402
10.1385/0896033198 doi
Hongos
Biotecnología
TP248.27 .Y43 / 1996 EB
Isolation and Identification of Yeasts from Natural Habitats -- Maintenance and Culture of Yeasts -- Mutagenesis in Yeast -- Rare-Mating and Cytoduction in Saccharomyces cerevisiae -- Protoplast Fusion in Saccharomyces cerevisiae -- Meiotic Analysis -- Ascus Dissection -- Karyotyping of Yeast Strains by Pulsed-Field Gel Electrophoresis -- Chromosomal Localization of Genes Through Pulsed-Field Gel Electrophoresis Techniques -- Isolation, Purification, and Analysis of Nuclear DNA in Yeast Taxonomy -- Isolation of Yeast DNA -- Isolation of Mitochondrial DNA -- Isolation of Yeast Plasma Membranes -- The Extraction and Analysis of Sterols from Yeast -- Peroxisome Isolation -- Transformation of Lithium-Treated Yeast Cells and the Selection of Auxotrophic and Dominant Markers -- Biolistic Transformation of Yeasts -- Genomic Yeast DNA Clone Banks -- Yeast Colony Hybridization -- Determination of Plasmid Copy Number in Yeast -- Determination of Chromosome Ploidy in Yeast -- Chromosome Engineering in Yeast with a Site-Specific Recombination System from a Heterologous Yeast Plasmid -- Insertional Mutagenesis by Ty Elements in Saccharomyces cerevisiae -- Reporter Gene Systems for Assaying Gene Expression in Yeast -- Preparation and Use of Yeast Cell-Free Translation Lysate -- Electrophoretic Analysis of Yeast Proteins -- Preparation of Total RNA -- mRNA Abundance and Half-Life Measurements -- Polysome Analysis -- Induction of Heat Shock Proteins and Thermotolerance -- Rhodamine B Assay for Estimating Activity of Killer Toxins Permeabilizing Cytoplasmic Membranes -- Nystatin-Rhodamine B Assay for Estimating Activity of Killer Toxin from Kluyveromyces lactis -- Application of Killer Toxins in Stepwise Selection of Hybrids and Cybrids Obtained by Induced Protoplast Fusion -- Killer Plaque Technique for Selecting Hybrids and Cybrids Obtained by Induced Protoplast Fusion -- Genotoxicity Testing in Schizosaccharomyces pombe -- Purification and Quantification of Saccharomyces cerevisiae Cytochrome P450 -- Calorimetry of Whole Yeast Cells -- Light Microscopy Methods -- Fluorescence Microscopy Methods -- Immunoelectron Microscopy.
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.
9781592595402
10.1385/0896033198 doi
Hongos
Biotecnología
TP248.27 .Y43 / 1996 EB