Click to have a closer look
About this book
This book describes and discusses experimental procedures used in a wide range of specialized topics within the genetics and molecular biology of the yeast Saccharomyces cerevisiae. Contributions are by internationally-recognized experts in particular areas and represent up-to-date methods currently in use. Topics covered include DNA isolation, cloning, and expression vectors, cosmid cloning, construction and use of cDNA libraries, plasmid shuffling and mutant isolation, Ty insertional mutagenesis, high efficiency transformation, cell-free translation of mRNAs, Ty virus-like particles, and applications to industrial strains of yeast. Detailed protocols are clearly presented, with many additional practical tips, to assist both first-time and more experienced experimentalists.
1. Procedures for isolating yeast DNA for different purposes; 2. Construction of cloning and expression vectors; 3. Cosmid cloning of yeast DNA; 4. The construction and use of cDNA libraries for genetic selections; 5. Pulsed field gel electrophoresis; 6. Plasmid shuffling and mutant isolation; 7. Ty insertional mutagenesis; 8. High efficiency transformation with lithium acetate; 9. Measurement of transcription; 10. Measurement of mRNA stability; 11. Production of foreign proteins at high level; 12. Cell-free translation of natural and synthetic mRNAs; 13. Virus-like particles: Ty retrotransposons; 14. The K1 killer toxin: molecular and genetic applications to secretion and cell surface assembly; 15. Immuno-electron microscopy; 16. Industrial Saccharomyces yeasts