Bernard R. Glick, Cheryl L. Patten

Molecular Biotechnology (eBook, PDF)

Principles and Applications of Recombinant DNA

• Format: PDF

Produktbeschreibung

Molecular Biotechnology Molecular Biotechnology Principles and Applications of Recombinant DNA SIXTH EDITION An authoritative introduction to the fast-changing world of molecular biotechnology In continuous publication since 1994 and now in its sixth edition, Molecular Biotechnology: Principles and Applications of Recombinant DNA has been effective in introducing this complex field to students for more than 25 years. This textbook covers essentially every aspect of the field of molecular biotechnology, which is constantly changing and adapting in light of new advances. This edition includes the latest techniques in DNA sequencing and genetic engineering of microbial, plant, and animal genomes, including human genome editing, as well as updates across many areas, such as: * Immunological assays for disease diagnosis, more effective bacteriophage therapy, and new ways of dealing with antibiotic-resistant bacteria * New and developing vaccines for influenza, tuberculosis, and emerging viral threats, including Zika and SARS-CoV-2 * Engineering bacteria to perform plastic degradation and green algae to produce hydrogen, altering amino acid biosynthesis, and creating designer cellulosomes * Production of humanized monoclonal antibodies in plants, modifying hybrid plants to produce clonal hybrids, and protecting plants from viral and fungal diseases Molecular Biotechnology features nearly 600 detailed figures and is an ideal textbook for undergraduate and graduate courses in introductory biotechnology, as well as courses dedicated to utilizing this technology, such as medical, agricultural, environmental, and industrial biotechnology applications.

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Produktdetails

• Verlag: John Wiley & Sons
• Seitenzahl: 896
• Erscheinungstermin: 31. Januar 2022
• Englisch
• ISBN-13: 9781683673651
• Artikelnr.: 63398028

Autorenporträt

About the Authors Bernard R. Glick, PhD, is a distinguished professor emeritus at the University of Waterloo, Ontario, Canada, where he was the chair of biology from 2002 to 2008. He has authored more than 400 publications that have garnered over 54,000 citations. Cheryl L. Patten, PhD, is a professor of microbiology in the Biology Department at the University of New Brunswick, Fredericton, Canada. She teaches courses in introductory biology, microbiology, and biotechnology.

Inhaltsangabe

1.The Development of Molecular Biotechnology 1

Emergence of Molecular Biotechnology 1

Recombinant DNA Technology 3

Commercialization of Molecular Biotechnology 6

Concerns and Consequences 8

Summary 10

References 10

Review Questions 11

2.Fundamental Technologies 13

Molecular Cloning 13

Preparation of DNA for Cloning 13

Insertion of Target DNA into a Plasmid Vector 18

Transformation and Selection of Cloned DNA in a Bacterial Host 21

Cloning Eukaryotic Genes 26

Recombinational Cloning 30

Genomic Libraries 32

Genome Engineering Using CRISPR Technology 36

Polymerase Chain Reaction 39

Amplification of DNA by PCR 39

Cloning PCR Products 42

Quantitative PCR 44

Chemical Synthesis of Genes 46

Assembling Oligonucleotides into Genes 46

Assembling PCR Products into Genes 46

DNA Sequencing Technologies 48

Dideoxynucleotide Sequencing 50

Sequencing Using Reversible Chain Terminators 53

Single-Molecule Real-Time Sequencing 55

Nanopore Sequencing 56

Sequencing Whole Genomes 56

Preparation of Genomic DNA Sequencing Libraries 57

High-Throughput Next-Generation Sequencing 59

Genome Sequence Assembly 60

Sequencing Metagenomes 61

Genomics 62

Transcriptomics 65

Proteomics 70

Metabolomics 84

Summary 86

References 87

Review Questions 89

3.Production of Recombinant Proteins 91

Protein Production in Prokaryotic Hosts 91

Regulating Transcription 92

Increasing Translation Efficiency 96

Increasing Protein Stability 100

Increasing Protein Secretion 105

Facilitating Protein Purification 111

Integrating DNA into the Host Chromosome 114

Heterologous Protein Production in Eukaryotic Cells 119

Posttranslational Modification of Eukaryotic Proteins 120

General Features of Eukaryotic Expression Systems 123

Yeast Expression Systems 123

Baculovirus-Insect Cell Expression Systems 137

Mammalian Cell Expression Systems 145

Protein Engineering 156

Directed Mutagenesis 156

Random Mutagenesis 161

Examples of Protein Engineering 164

Summary 173

References 174

Review Questions 176

4.Molecular Diagnostics 179

Immunological Approaches To Detect Protein Biomarkers 180

Antibodies 180

Agglutination 185

Enzyme-Linked Immunosorbent Assays 186

Protein Arrays To Detect Polygenic Diseases 194

Immunoassays for Protein Conformation-Specific Disorders 197

DNA-Based Diagnostic Approaches 199

Hybridization Probes 199

PCR-Based Detection Methods 207

CRISPR-Cas-Based Diagnostic Assays 218

DNA Microarrays 219

Whole-Genome Sequencing To Assess Genetic Disease Risk 225

Detecting RNA Signatures of Disease 226

Detection of Disease-Associated Changes in Gene Expression 227

Detection of RNA Signatures of Antibiotic Resistance in Bacteria 228

Detection of miRNA Signatures of Disease 230

Biofluorescent and Bioluminescent Systems 233

Fluorescent Proteins 233

Luciferase 234

Microbial Biosensors 235

Summary 238

References 239

Review Questions 241

5.Protein Therapeutics 243

Pharmaceuticals 244