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.
Author(s): Bernard R. Glick, Cheryl L. Patten
Edition: 6
Publisher: Wiley
Year: 2022
Language: English
Pages: 899
Cover
Title Page
Copyright Page
About the Companion Website
Contents
Preface to the Sixth Edition
Chapter 1 The Development of Molecular Biotechnology
Emergence of Molecular Biotechnology
Recombinant DNA Technology
Commercialization of Molecular Biotechnology
Concerns and Consequences
summary
REFERENCES
review questions
Chapter 2 Fundamental Technologies
Molecular Cloning
Preparation of DNA for Cloning
Insertion of Target DNA into a Plasmid Vector
Transformation and Selection of Cloned DNA in a Bacterial Host
Cloning Eukaryotic Genes
Recombinational Cloning
Genomic Libraries
Genome Engineering Using CRISPR Technology
Polymerase Chain Reaction
Amplification of DNA by PCR
Cloning PCR Products
Quantitative PCR
Chemical Synthesis of Genes
Assembling Oligonucleotides into Genes
Assembling PCR Products into Genes
DNA Sequencing Technologies
Dideoxynucleotide Sequencing
Sequencing Using Reversible Chain Terminators
Single‐Molecule Real‐Time Sequencing
Nanopore Sequencing
Sequencing Whole Genomes
Preparation of Genomic DNA Sequencing Libraries
High‐Throughput Next‐Generation Sequencing
Genome Sequence Assembly
Sequencing Metagenomes
Genomics
Transcriptomics
Proteomics
Metabolomics
summary
REFERENCES
review questions
Chapter 3 Production of Recombinant Proteins
Protein Production in Prokaryotic Hosts
Regulating Transcription
Increasing Translation Efficiency
Increasing Protein Stability
Increasing Protein Secretion
Facilitating Protein Purification
Integrating DNA into the Host Chromosome
Heterologous Protein Production in Eukaryotic Cells
Posttranslational Modification of Eukaryotic Proteins
General Features of Eukaryotic Expression Systems
Yeast Expression Systems
Baculovirus–Insect Cell Expression Systems
Mammalian Cell Expression Systems
Protein Engineering
Directed Mutagenesis
Random Mutagenesis
Examples of Protein Engineering
summary
REFERENCES
review questions
Chapter 4 Molecular Diagnostics
Immunological Approaches To Detect Protein Biomarkers
Antibodies
Agglutination
Enzyme‐Linked Immunosorbent Assays
Protein Arrays To Detect Polygenic Diseases
Immunoassays for Protein Conformation‐Specific Disorders
DNA‐Based Diagnostic Approaches
Hybridization Probes
PCR‐Based Detection Methods
CRISPR‐Cas‐Based Diagnostic Assays
DNA Microarrays
Whole‐Genome Sequencing To Assess Genetic Disease Risk
Detecting RNA Signatures of Disease
Detection of Disease‐Associated Changes in Gene Expression
Detection of RNA Signatures of Antibiotic Resistance in Bacteria
Detection of miRNA Signatures of Disease
Biofluorescent and Bioluminescent Systems
Fluorescent Proteins
Luciferase
Microbial Biosensors
summary
REFERENCES
review questions
Chapter 5 Protein Therapeutics
Pharmaceuticals
Human Interferons
Human Growth Hormone
Tumor Necrosis Factor Alpha
Extending Protein Half-Life
Enzymes
DNase I
Alginate Lyase
Phenylalanine Ammonia Lyase
1-Antitrypsin
Glycosidases
Masking Nonhuman Epitopes
Toxin-Intein Fusions
Targeting Mitochondria
Bacteria and Therapeutics
Interleukin-10
Leptin
An HIV Inhibitor
Insulin
Parkinson’s Disease
Cancer and Bacteria
Recombinant Antibodies
Hybrid Human–Mouse Monoclonal Antibodies
Human Monoclonal Antibodies
Antibody Fragments
Combinatorial Libraries of Antibody Fragments
A Combinatorial Library of Full-Length Antibodies
Shuffling CDR Sequences
Dual-Variable-Domain Antibodies
Bispecific Antibodies against Hemophilia
Anti-HIV Antibodies
Anticancer Antibodies
Antibodies against Various Diseases
Antiobesity Antibodies
Enhanced Antibody Half-Life
Affibody Molecules
summary
References
review questions
Chapter 6 Nucleic Acids as Therapeutic Agents
Targeting Specific mRNA and DNA Sequences
Antisense RNA
Aptamers
Ribozymes and DNAzymes
Interfering RNA
Zinc Finger Nucleases
CRISPR‐Cas System
Nanozymes
Nanoparticles
Engineering Bacteriophages
Viral Delivery Systems
Nonviral Delivery Systems
Direct Injection
Lipids
Bacteria
Dendrimers
Antibodies
Aptamers
Transposons
Gene Therapy
Mitochondrial Diseases
Prodrug Activation Therapy
Promoterless Gene Targeting
summary
References
review questions
Chapter 7 Vaccines
Vaccination
Current and Future Vaccines
Subunit and Peptide Vaccines
Herpes Simplex Virus
Bovine Herpes Virus-1
Cholera
Influenza
SARS
COVID-19
Staphylococcus aureus
Human Papillomavirus
Foot-and-Mouth Virus
Streptococcus
Peptides
Malaria
Delivery
Genetic Immunization: DNA Vaccines
Delivery
Cancer
Zika Virus
Dental Caries
Engineered Attenuated Vaccines
Herpes Simplex Virus
Cholera
Salmonella Species
Leishmania Species
Vector Vaccines
Vaccines Directed against Viruses
Vaccines Directed against Bacteria
Bacteria as Antigen Delivery Systems
Monoclonal Antibody Passive Immunity
Influenza Virus
summary
References
review questions
Chapter 8 Industrial and Environmental Uses of Recombinant Microorganisms
Restriction Endonucleases
Small Biological Molecules
l-Ascorbic Acid
Indigo
Amino Acids
Lycopene
Antibiotics
Biopolymers
Solvent Tolerance
Systems Metabolic Engineering To Optimize Product Yield
Microbial Degradation of Xenobiotics
Genetic Engineering of Biodegradative Pathways
Plastics
Utilization of Starch and Sugars
Commercial Production of Fructose and Alcohol
Increasing Alcohol Production
Improving Fructose Production
Utilization of Cellulose and Hemicellulose
Lignocellulosics
Cellulase Genes
Direct Conversion of Biomass to Ethanol
Alcohol Production by Zymomonas mobilis
Lipids from Cyanobacteria
Hydrogen Production
summary
References
review questions
Chapter 9 Large-Scale Production of Proteins and Nucleic Acids from Recombinant Microorganisms
Principles of Microbial Growth
Batch Fermentation
Fed-Batch Fermentation
Continuous Fermentation
Maximizing the Efficiency of the Fermentation Process
High-Density Cell Cultures
Increasing Plasmid Stability
Quiescent E. coli Cells
Protein Secretion
Reducing Acetate
Improving Antibody Production in E. coli
Bioreactors
Typical Large-Scale Fermentation Systems
Two-Stage Fermentation in Tandem Airlift Reactors
Two-Stage Fermentation in a Single Stirred-Tank Reactor
Batch versus Fed-Batch Fermentation
Harvesting Microbial Cells
Disrupting Microbial Cells
Downstream Processing
Inclusion Bodies
Utilizing an Immobilized Enzyme
Magnetic Separation of Proteins
Large-Scale Production of DNA and RNA
Plasmid DNA
mRNA
summary
References
review questions
Chapter 10 Genetic Engineering of Plants: Methodology
Plant Transformation with the Ti Plasmid of A. tumefaciens
Ti Plasmid-Derived Vector Systems
Increasing Transformation Efficiency
Microprojectile Bombardment
Chloroplast Engineering
Very-High-Level Protein Expression
Use of Reporter Genes in Transformed Plant Cells
Manipulation of Gene Expression in Plants
Transient Gene Expression
Plant Promoters
Manipulation of Genes in Plants
Facilitating Protein Purification
Protein Glycosylation
Gene Stacking
CRISPR-Based Directed Evolution
Polycistronic Gene Expression
Production of Marker-Free Transgenic Plants
Removing Marker Genes from Nuclear DNA
Removing Marker Genes from Chloroplast DNA
summary
References
review questions
Chapter 11 Transgenic Plants
Insect Resistance
Bacillus thuringiensis Insecticidal Toxin
Increasing Expression of the B. thuringiensis Protoxin
Other Strategies for Protecting Plants against Insects
Preventing the Development of B. thuringiensis-Resistant Insects
Targeting Aphids
Virus Resistance
Viral Coat Protein-Mediated Protection
Protection by Expression of Other Genes
Herbicide Resistance
Glyphosate
Dicamba
Other Herbicides
Fungus and Bacterium Resistance
Transgenic Plants
RNAi and CRISPR/Cas
Salt and Drought Stress
Increasing Trehalose Production
Sequestering Sodium Ions
Delaying Drought-Induced Senescence
Phytoremediation
Fruits and Flowers
Flavr Savr Tomato
Lowering Ethylene Levels
CRISPR Mutants
Modification of Plant Nutritional Content
Amino Acids
Lipids
Vitamins
Iron
Gluten
Modification of Food Plant Taste and Appearance
Preventing Discoloration
Starch
Plants as Bioreactors
Antibodies
Pharmaceuticals and Vaccines
Poly(3-Hydroxybutyric Acid)
Edible Vaccines
Edible Cholera Vaccines
Edible E. coli Vaccines
Plant Yield
Increasing Grain Yield
Increasing Harvest Index
Decreasing Lignin Content
Decreasing Pectin Content
Increasing Oxygen Content
summary
References
review questions
Chapter 12 Transgenic Animals
Transgenic Animal Methodologies
DNA Microinjection Method
Retroviral Vector Method
Engineered Embryonic Stem Cell Method
Somatic Cell Nuclear Transfer for Transgenic Livestock
Genome Editing with the CRISPR‐Cas System
Conditional Gene Modification with the Cre–loxP Recombination System
Control of Transgene Expression with the Tetracycline‐Inducible System
Gene Knockdown by RNA Interference
Transgenic Animal Models of Human Diseases
Mouse Models of Alzheimer’s Disease
Mouse Model of Duchenne Muscular Dystrophy
Rabbit Models of Cardiovascular Disease
Zebrafish Melanoma Model
Nonhuman Primate Models of Neurodevelopmental Disorders
Animal Bioreactors for Production of Recombinant Therapeutic Proteins
Production of Recombinant Antithrombin in Goat Milk
Production of a Human Protease Inhibitor in Rabbits
Production of Therapeutic Proteins in Chicken Eggs
Production of Donor Organs in Pigs
Enhancing Production Traits of Food Animals
Disease‐Resistant Livestock
Improving Milk Quality
Increasing Muscle Mass in Cattle
Enhancing Growth of Salmon
Gene Drives To Eradicate Vector‐Transmitted Diseases
Malaria Vector Population Suppression
Dengue Fever Virus‐Resistant Mosquitoes
Reversal Drives
summary
REFERENCES
review questions
Chapter 13 Molecular Biotechnology and Society
Development of Guidelines for Recombinant DNA Research
Deliberate Release of Genetically Modified Microorganisms
Environmental Concerns
Regulations
Regulation of Genetically Modified Foods
Food Ingredients Produced by Genetically Engineered Microorganisms
Genetically Modified Crops
Genetically Engineered Livestock
Societal Concerns about Genetically Modified Foods
Alteration of Nutritional Content of Food
Potential for Introducing Toxins or Allergens into Food
Potential for Transferring Transgenes from Food to Humans or Intestinal Microorganisms
Controversy about the Labeling of Genetically Modified Foods
Impact of Genetically Engineered Crops on Biodiversity
Who Benefits from the Production of Genetically Modified Foods?
Environmental Benefits of Genetically Modified Crops
How Do Views about Genetically Engineered Organisms Impact Trade?
Regulation and Safety of Medical Products of Biotechnology
New Biological Drugs
Genetic and Genomic Testing
Economic Issues
Patenting Biotechnology
Patenting
Patenting in Different Countries
Patenting Nucleic Acid Sequences
Patenting Living Organisms
Patenting and Fundamental Research
summary
REFERENCES
review questions
Amino Acids of Proteins and Their Designations
Index
EULA
