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This manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students gain hands-on experience from start to finish in subcloning a gene into an expression vector, through purification of the recombinant protein. The second edition has been completely re-written, with new laboratory exercises and all new illustrations and text, designed for a typical 15-week semester, rather than a 4-week intensive course. The "project" approach to experiments was maintained: students still follow a cloning project through to completion, culminating in the purification of recombinant protein. It takes advantage of the enhanced green fluorescent protein-students can actually visualize positive clones following IPTG induction. *Cover basic concepts and techniques used in molecular biology research labs *Student-tested labs proven successful in a real classroom laboratories *Exercises simulate a cloning project that would be performed in a real research lab *"Project" approach to experiments gives students an overview of the entire process *Prep-list appendix contains necessary recipes and catalog numbers, providing staff with detailed instructions.

Molecular biology --- Molecular biology --- DNA, Recombinant --- Molecular Biology --- Protein Engineering --- Technique --- methods

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Synthetic biology --- Genetic engineering --- Synthetic Biology. --- Genetic Engineering. --- Genetic engineering. --- Synthetic biology. --- Designed genetic change --- Engineering, Genetic --- Gene splicing --- Genetic intervention --- Genetic surgery --- Intervention, Genetic --- Genetic Intervention --- Genetic Interventions --- Interventions, Genetic --- Biologies, Synthetic --- Biology, Synthetic --- Synthetic Biologies --- Biology --- Engineering --- Bioengineering --- Genetic recombination --- Biotechnology --- Transgenic organisms --- Cloning, Molecular --- DNA, Recombinant --- Industrial Microbiology --- Artificial Gene Fusion --- Organisms, Genetically Modified --- Animals, Genetically Modified --- Plants, Genetically Modified --- Synthetic Biology --- Genetic Engineering

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In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly 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 laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.   Authoritative and practical, Recombinant Protein Production in Yeast: Methods and Protocols, seeks to aid scientists in adopting yeast as a protein production host.

Pichia --- Recombinant Proteins --- Recombinant proteins --- Saccharomyces cerevisiae --- Yeast fungi --- Yeasts --- biosynthesis --- Biotechnology --- Fungi --- Proteins --- Publication Formats --- Saccharomyces --- Saccharomycetales --- Publication Characteristics --- Ascomycota --- Eukaryota --- Amino Acids, Peptides, and Proteins --- Chemicals and Drugs --- Organisms --- Laboratory Manuals --- Health & Biological Sciences --- Biomedical Engineering --- Baker Yeast --- S cerevisiae --- Baker's Yeast --- Brewer's Yeast --- Yeast, Baker's --- Yeast, Brewer's --- Baker Yeasts --- Baker's Yeasts --- Bakers Yeast --- Brewer Yeast --- Brewer's Yeasts --- Brewers Yeast --- Yeast, Baker --- Yeast, Bakers --- Yeast, Brewer --- Yeast, Brewers --- Yeasts, Baker --- Yeasts, Baker's --- Yeasts, Brewer's --- Proteins, Biosynthetic --- Proteins, Recombinant DNA --- Biosynthetic Proteins --- DNA Recombinant Proteins --- DNA Proteins, Recombinant --- Proteins, DNA Recombinant --- Proteins, Recombinant --- Recombinant DNA Proteins --- Recombinant Proteins, DNA --- Hansenula --- Hansenulas --- Pichias --- Yeast --- Genetically engineered proteins --- Eucarya --- Eukarya --- Eukaryotes --- Eukaryotas --- Eukaryote --- Ascomycetes --- Cochliobolus --- Sclerotinia --- Ascomycete --- Ascomycotas --- Sclerotinias --- Budding Yeast --- Endomycetales --- Endomycopsis --- Yeast, Budding --- Budding Yeasts --- Endomycetale --- Endomycopses --- Saccharomycetale --- Yeasts, Budding --- Gene Products, Protein --- Gene Proteins --- Protein Gene Products --- Proteins, Gene --- Fungi, Filamentous --- Molds --- Filamentous Fungi --- Filamentous Fungus --- Fungus --- Fungus, Filamentous --- Mold --- Genetic engineering --- Saccharomyce --- Medicine. --- Human genetics. --- Biomedicine. --- Human Genetics. --- Genetics --- Heredity, Human --- Human biology --- Physical anthropology --- Clinical sciences --- Medical profession --- Life sciences --- Medical sciences --- Pathology --- Physicians