Recombinant DNA:
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| Format: | Buch |
|---|---|
| Sprache: | Undetermined |
| Veröffentlicht: |
New York
Freeman
1992
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| Ausgabe: | 2. ed. |
| Schriftenreihe: | Scientific American books
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XII, 626 S. Ill., graph. Darst. |
| ISBN: | 0716722828 |
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Datensatz im Suchindex
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| adam_text | Recombinant DNA
SECONDEDITION
James D Watson
COLDSPRINGHARBORLABORATORY
Michael Gilman
COLDSPRINGHARBORLABORATORY
Jan Witkowski
BANBURYCENTER,COLDSPRINGHARBORLABORATORY
Mark Zoller
GENENTECH,INC
SCIENTIFIC
AMERICAN BOOKS
Distributed by
W H Freeman and Company
New York
Contents
Preface xiii
Acknowledgments xiv
Development of Recombinant DNA Technology
1 ESTABLISHING THE ROLE OF GENES WITHIN CELLS 1
The Building Blocks of All Life Are Cells 2
Cells Are Tiny Expandable Factories That Simultaneously
Synthesize Several Thousand Different Molecules 2
A Cell s Molecules Can Be Divided into Small Molecules and
Macromolecules 3
Special Cellular Catalysts Called Enzymes Effectively Determine the
Chemical Reactions That Occur in Cells 3
A Given Protein Possesses a Unique Sequence of Amino Acids
Along Its Polypeptide Chain 3
The Functioning of an Enzyme Demands a Precise Folding of Its
Polypeptide Chain 4
Activation of Molecules to High-Energy Forms Promotes Their
Chemical Reactivity 5
Cellular Metabolism Can Be Visualized Through Metabolic Maps 6
Enzymes Cannot Determine the Order of Amino Acids in Polypeptide
Chains 6
hCONTENTS
Mendel s Experiments with Pea Plants First
Revealed the Discreteness of Genetic
Determinants (Genes) 6
Chromosomes Are the Cellular Bearers of
Heredity 1
The One Gene, One Protein Hypothesis Is
Developed 10
2 DNA IS THE PRIMARY GENETIC
MATERIAL 13
DNA Is Sited on Chromosomes 13
Cells Contain RNA as well as DNA 14
A Biological Assay for Genetic Molecules Is
Discovered 15
Viruses Are Packaged Genetic Elements That
Move from Cell to Cell 16
Molecules with Complementary Sizes and
Shapes Attract One Another 17
The Diameter of DNA Is Established 18
The Nucleotides of DNA and RNA Are
Linked Together by Regular 5 -3
Phosphodiester Bonds 18
The Composition of Bases of DNA from
Different Organisms Varies Greatly 19
DNA Has a Highly Regular Shape 19
The Fundamental Unit of DNA Consists of
Two Intertwined Polynucleotide Chains
(the Double Helix) 20
The Double Helix Is Held Together by
Hydrogen Bonds Between Base Pairs 21
The Complementary Nature of DNA Is at the
Heart of Its Capacity for Self-Replication 22
DNA Replication Is Found to Be
Semiconservative 23
DNA Molecules Can Be Renatured as well as
Denatured 23
G C Ba sePairsFallApartLessEasilyThan
Their A*T Equivalents 24
Palindromes Promote Intrastrand Hydrogen
Bonding 24
5-Methylcytosine Can Replace Cytosine in
DNA 24
Chromosomes Contain Single DNA Molecules 25
Viruses Are Sources of Homogeneous DNA
Molecules 25
Phage 1 DNA Can Insert into a Specific Site
Along the E coli Chromosome 26
Abnormal Transducing Phages Provide Unique
Segments of Bacterial Chromosomes 26
Plasmids Are Autonomously Replicating
Minichromosomes 27
Circular DNA Molecules May Be Supercoiled 28
Most Double Helices Are Right-Handed,
but Under Special Conditions Certain
DNA Nucleotide Sequences Lead to
Left-Handed Helices 30
3 ELUCIDATION OF THE GENETIC CODE 33
A Mutation in the Hemogloblin Molecule
Is Traced to a Single Amino Acid
Replacement 33
The Use of K Coli Leads to the Rapid
Development of Fine-Structure Genetics 34 -
The Gene and Its Polypeptide Products Are
Colinear 35
RNA Carries Information from DNA to the
Cytoplasmic Sites of Protein Synthesis 35
How Do Amino Acids Line Up on RNA
Templates? 36
Roles of Enzymes and Templates in the
Synthesis of Nucleic Acids and Proteins 31
Proteins Are Synthesized from the Amino
Terminus to the Carboxyl Terminus 31
Three Forms of RNA Are Involved in Protein
Synthesis 31
Generic Evidence Reveals That Codons
Contain Three Bases 39
RNA Chains Are Both Synthesized and
Translated in a 5 -to-3 Direction 39
Synthetic mRNA Is Used to Make the Codon
Assignments 41
The Genetic Code Is Fully Deciphered by
June 1966 41
Wobble Frequently Permits Single tRNA
Species to Recognize Multiple Codons 42
How Universal Is the Genetic Code? 42
Average-Sized Genes Contain at Least 1200
Base Pairs 42
Mutations Change the Base Sequence of DNA 43
Suppressor tRNAs Cause Misreading of the
Genetic Code 43
The Signals for Starting and Stopping the
Synthesis of Specific RNA Molecules Are
Encoded Within DNA Sequences 44
Increasingly Accurate Systems Are Developed
for the in Vitro Translation of
Exogenously Added mRNAs 45
4 THE GENETIC ELEMENTS THAT
CONTROL GENE EXPRESSION 49
Repressors Control Inducible Enzyme
Synthesis 50
Contents V
Bacterial Genes with Related Functions Are
Organized into Operons 51
Promoters Are the Start Signals for RNA
Synthesis 51
Repressor Molecules Are Normally Made at
Constant Rates 52
Repressors Are Isolated and Identified 52
Positive Regulation of Gene Transcription Also
Occurs 53
Attenuation Is Another Form of Regulation 53
Translational Control Is the Second Means of
Controlling Protein Synthesis 54
Probing Gene Regulation in Higher Plants and
Animals Presented Early Difficulties 55
Purified Xenopus Ribosomal RNA Genes Are
Isolated 56
Eukaryotic mRNAs Have Caps and Tails 56
Eukaryotes Are Found to Have Three Kinds of
RNA Polymerases 57
Eukaryotic DNA Is Organized into
Nucleosomes 57
Animal Viruses Are Model Systems for Gene
Expression in Higher Cells 58
The RNA Tumor Viruses Replicate by Means
of a Double-Stranded DNA Intermediate 58
5 METHODS OF CREATING
RECOMBINANT DNA MOLECULES 63
Nucleic Acid Sequencing Methods Are
Developed 63
Restriction Enzymes Make Sequence-Specific
Cuts in DNA _ 64
Restriction Maps Are Highly Specific - 66
Restriction Fragments Lead to Powerful New
Methods for Sequencing DNA 67
Oligonucleotides Can Be Synthesized
Chemically 69
Many Restriction Enzymes Produce Fragments
Containing Sticky (Cohesive) Ends 70
Many Enzymes Are Involved in DNA
Replication 71
Sticky Ends May Be Enzymatically Added to
Blunt-Ended DNA Molecules 72
Small Plasmids Are Vectors for the Cloning of
Foreign Genes 73
DNA of Higher Organisms Becomes Open to
Molecular Analysis 74
Scientists Voice Concerns About the Dangers
of Unrestricted Gene Cloning 75
Guidelines for Recombinant DNA Research
Are Proposed at the Asilomar Conference 75
Recombinant DNA Comes of Age 75
Analysis of Cloned Genes
6 THE POLYMERASE CHAIN REACTION 79
The Polymerase Chain Reaction Amplifies
Specific Regions of DNA 79
Performing a Polymerase Chain Reaction 80
Tag Polymerase Simplifies and Improves the
PCR 82
Fidelity of DNA Synthesis by Taq Polymerase
Determines the Accuracy of PCR
Amplification 84
DNA for the PCR Comes from a Variety of
Sources 85
PCR is Used to Amplify Human-Specific DNA
Sequences 86
PCR Products Can Be Sequenced Directly 88
Detecting Mutations Using PCR Amplification 88
PCR Amplification Is Used for Monitoring
Cancer Therapy 89
PCR Amplification Is Used to Detect Bacterial
and Viral Infections 89
PCR Amplification Is Used For Sex
Determination of Prenatal Cells 91
PCR Methods Permit Linkage Analysis Using
Single Sperm Cells 92
PCR Techniques Are Used in Studies of
Molecular Evolution 94
Contamination Can Be a Problem in PCR
Studies 95
The Polymerase Chain Reaction—A Technical
Revolution in Molecular Genetics 95
7 THE ISOLATION OF CLONED GENES 99
Improved Bacteria and Vectors Are Developed 100
Basic Strategies for Cloning Involve Three
Steps 100
Choosing the Right Starting Material Is
Essential in Cloning 102
mRNA Is Converted to cDNA by Enzymatic
Reactions 102
cDNA Molecules Are Joined to Vector DNA
for Propagation in Bacteria 104
Nucleic Acid Probes Are Used to Locate
Clones Carrying a Desired DNA Sequence 104
Synthetic Oligonucleotide Probes Can Be
Designed from Known Protein Sequence 107
Tissue-Specific cDNAs Are Identified by
Differential Hybridization 107
Gene Probes from Conserved Segments in
Protein Families Identify New Related
Genes 111
vi CONTENTS
Expression Vectors May Be Used to Isolate
Specific cDNAs 113
Cloned Genes Can Be Isolated by Functional
Assay in E coli 113
Cloned Genes Can Be Isolated by Functional
Assay in Eukaryotic Cells 115
Cloned DNA Is Analyzed by DNA Sequencing 116
Computers Have Simplified Translating DNA
Sequence into Protein Sequence 119
Searching Sequence Databases to Identify
Proteins and Protein Functions 120
Several Procedures Exist to Analyze Proteins
Encoded by cDNA Clones 121
Genomic Fragments Are Cloned in
Bacteriophage 124
Cosmids Allow the Cloning of Large Segments
of Genomic DNA 127
Chromosome Walking Is Used to Analyze
Long Stretches of Eukaryotic DNA 127
Southern and Northern Blotting Procedures
Analyze DNA and RNA 127
8 THE COMPLEXITY OF THE GENOME 135
Split Genes Are Discovered 136
Introns Are Discovered in Eukaryotic Genes 137
Specific Base Sequences Are Found at Exon-
Intron Boundaries 138
Alternative Splicing Pathways Generate
Different mRNAsJrom a Single Gene 140
Introns Sometimes Mark Functional Protein
Domains 140
Transcriptional Control Regions Occur
Throughout Eukaryotic Genes 142
RNA Polymerase III Transcription Is
Regulated by Sequences in the Middle of
the Gene 143
Genes Encoding Abundant Products Are Often
Tandemly Repeated 143
Clustered Globin Genes Exhibit Coordinated
Expression in Development 144
Pseudogenes Arise by Duplication of an Active
Gene and Accumulate Mutations During
Evolution 145
Short Repetitive DNA Sequences Are
Dispersed Throughout the Eukaryotic
Genome 146
Transcriptionally Inactive Regions of the
Genome Are Often Found to Be
Methylated 147
Most Genes Are Much Larger Than Their
mRNA 147
Genes Are Sometimes Encoded Within Genes 148
Summary 149
9 CONTROLLING EUKARYOTIC GENE
EXPRESSION 153
Common Sequence Motifs Are Recognized in
5 -Flanking Regions 153
Enhancers Activate Gene Expression over Long
Distances 155
Enhancers Can Be Tissue-Specific or
Regulated by Signals 157
Enhancers Contain Recognition Sites for DNA-
Binding Proteins 158
Genes Can Be Transcribed in Cell-Free
Extracts 159
Gene-Specific Transcription Factors Are DNA-
Binding Proteins 160
Transcription Factors Are Purified and Cloned 161
Transcription Factors Fall into Structural
Families 163
Transcription Factors Are Modular 165
How Do Transcription Factors Work? 167
The Cellular Splicing Machinery Is
Extraordinarily Selective and Precise 168
Synthetic Pre-mRNAs Are Spliced in Oocytes
and Cell-Free Extracts 169
Cell-Free Extracts Are Fractionated to Identify
Splicing Factors 170
Trans-Acting Splicing Factors Govern
Alternative Splicing 171
10 MOVABLE GENES 175
Sequencing Reveals the Organization of
Bacterial Transposable Elements 176
Some Transposable Elements in Eukaryotes
Resemble Bacterial Transposons 177
Ty Elements in Yeast Are a Different Class of
Transposable Elements 177
Repetitive Elements and Processed
Pseudogenes Are Remnants of
Transposition Events 179
Bacterial Transposons Jump via DNA
Intermediates 179
Ty Elements Transpose via an RNA
Intermediate 181
Mating Type Interconversion in Yeast Occurs
by Replicative Transposition 181
Functional Immunoglobulin Genes Are Formed
by Ordered Gene Rearrangements 183
Gene Rearrangements Sometimes Go Awry 185
Contents vii
Transposable Elements Are Potent Tools for
Identifying and Manipulating Genes 186
New Tools for Studying
Gene Function
11 IN VITRO MUTAGENESIS 191
In Vitro Mutagenesis Is Used to Study Gene
Function 192
Restriction Endonuclease Sites Provide the
Simplest Access for Mutagenesis 193
Linker Insertion Is Used to Map a Bacterial
Transposon 195
Construction of Nested Deletions Maps the
Boundaries of a Transcriptional Control
Region 195
Linker-Scanning Mutagenesis Permits
Systematic Analysis of Promoters 195
Random Nucleotide Substitutions Are
Obtained by Chemical Modification of
DNA or by Enzymatic Misincorporation 197
Synthetic Oligonucleotides Facilitate
Mutagenesis 201
Mutant Clones Can Be Identified by
Hybridization and DNA Sequencing 201
Oligonucleotide Cassettes Provide a Simple
Method for Introducing Directed
Mutations 202
Gene Synthesis Facilitates Production of
Normal and Mutant Proteins 204
The PCR Can Be Used to Construct Genes
Encoding Chimeric Proteins 207
Mutagenesis Is the Gateway to Gene Function
and Protein Engineering 209
12 TRANSFERRING GENES INTO
MAMMALIAN1 CELLS 213
Establishment of Immortal Cell Lines Makes
Gene Transfer Practical 214
Gene Transfer Was First Used in the Study of
Tumor Viruses 214
Selectable Markers That Work in Mammalian
Cells Allow Gene Transfer by
Cotransformation 216
Exogeneous DNA Is Transiently Expressed in
Many Cells Immediately Following
Transfection 218
Gene Amplification Is Used to Achieve High-
Level Protein Expression 219
Specialized Methods are Developed to
Transfect Difficult Cell Types 221
Viral Vectors Introduce Foreign DNA into
Cells with High Efficiency 222
Vaccinia and Baculovirus Are Used for High-
Level Protein Production 224
Retroviruses Provide High-Efficiency Vectors
for Stable Gene Transfer 225
The Experimenter Has Little Control over the
Fate of Transferred DNA 227
Antisense RNA and DNA Can Extinguish
Gene Function 228
Homologous Recombination Is Used to
Inactivate Cellular Genes 228
13 USING YEAST TO STUDY
EUKARYOTIC GENE FUNCTION 235
Yeast Biosynthetic Genes Are Cloned by
Complementation of E coli Mutations 236
Shuttle Vectors Replicate in Both E Coli and
Yeast 237
Yeast Genes Can Be Cloned by Simple
Complementation Strategies 239
Homologous Recombination Is a Relatively
Frequent Event in Yeast 240
Cloning Genes Required for Mating Reveals a
Signaling Pathway Similar to That Seen in
Higher Organisms 241
Genetic Experiments in Yeast Can Answer
Precise Biochemical Questions 245
Genetic Analysis in Yeast Can Be Exploited to
Identify and Study Genes from Higher
Organisms 250
14 THE INTRODUCTION OF FOREIGN
GENES INTO MICE 255
Foreign Genes Become Integrated in the
Chromosomes of Recipient Animals 256
Foreign DNA Can Become Stably Integrated
into Germ Line Cells 257
Embryonic Stem Cells Can Carry Foreign
Genes 257
Transgenes Can Be Regulated in a Tissue-
Specific Manner 259
Transgene Expression Can Be Targeted to
Specific Tissues 259
Transgenes Can Be Used to Kill Specific Cell
Types 260
Retroviruses Can Be Used to Trace Cell
Lineages 261
Transgenes Can Disrupt the Functioning of
Endogenous Genes 261
Viii CONTENTS
Knocking Out Genes by Homologous
Recombination Can Elucidate Complex
Gene Systems 261
Studying Genetic Control of Mouse
Development 263
Transgenes Can Be Used to Detect Host
Genes 263
A Single Gene Turns Female Mice into Males 264
Transgenic Mice Provide Models of Human
Diseases 266
Imprinting—Males and Females Make
Unequal Genetic Contributions to Their
Offspring 2 67
IS GENETIC ENGINEERING OF PLANTS 273
Plants Have Advantages and Disadvantages for
Genetic Engineering 274
Whole Plants Can Be Grown from Single Cells 274
Leaf Disks Are an Important Target for Gene
Transfer 275
Ti Plasmid of Agrobacterium Causes Crown Gall
Tumors 277
T-DNA, Part of the Ti Plasmid, Is Transferred
to Plant Cells 277
T-DNA Has Been Modified to Act as a Gene
Vector 278
Reporter Genes Demonstrate Transgene
Expression in Plant Tissues 281
Viruses Can Be Used as Vectors for Whole
Plants 282
Guns and Electric Shocks Transfer DNA into
Plant Cells — 282
Bombardment with DNA-Coated~Beads Can
Produce Transgenic Organelles 285
Plant Genes Can Be Cloned by Using
Transposable Elements 285
T-DNA Is Used as an Insertion Mutagen 287
Arabidopsis Is Being Used as a Model Organism
for Molecular Genetic Analysis of Plants 290
Analysis of Important Biological
Processes by Using Recombinant DNA
16 MOLECULES OF IMMUNE
RECOGNITION 293
The Basic Structure of Antibody Molecules Is
Established 294
Fledgling Recombinant DNA Technology
Verifies the Dreyer and Bennett
Hypothesis 295
Rearrangement of Antibody Genes Generates ,
Additional Diversity at the V-C Junction 295
Class Switching Places Useful Recognition
Specificities on Antibody Molecules with
Different Functional Properties 299
The Mechanisms of Antibody Gene
Rearrangement Can Be Studied by
Introducing Artificial Genes into Cells 300
The Study of Cellular Immunity Was Greatly
Advanced by Gene Cloning 302
T Cells Recognize Antigens Only on Cells
from the Same Individual 304
Cloning of the MHC Genes Reveals That Self
Identity is Determined by a Few
Polymorphic Genes 305
T-Cell Receptors Recognize Antigens Only in
Association with an MHC Molecule 305
Intercellular Communication Regulates
Immune System Function 307
The Immunoglobulin Superfamily Encodes
Proteins That Participate in Cell-Cell
Communication 309
Malfunctions of the Immune System Underlie
Many Diseases 310
17 MOVING SIGNALS ACROSS
MEMBRANES 313
The Acetylcholine Receptor Is a Ligand-Gated
Ion Channel 315
Some Receptors Are Coupled to Second
Messenger Systems via GTP-Binding
Proteins 315
G Protein-Linked Receptors Span the
Membrane Seven Times 317
Domain Swapping Reveals the Structural Basis
of Receptor-Effector Coupling 317
Visual Pigments Are Signaling Receptors 318
Growth Factor Receptors Have Intrinsic
Enzymatic Activity
Receptors Can Be Associated with Intracellular
Protein Kinases
Receptors Activate Common Second
Messenger Systems
Protein Phosphorylation Is a Principal
Mechanism for Signaling 325
Steroid Receptors Are Transcription Factors 327
cAMP Signals Reach the Nucleus via
Transcription Factor Phosphorylation 328
Studying Target Genes Reveals the
Organization of Signal Transduction
Pathways 329
Immediate Early Genes Are Third Messengers 331
Contents IX
18 ONCOGENES AND ANTI-ONCOGENES 335
Cancer Is a Genetic Disease 336
Tumor Cells Have Aberrant Growth Properties
in Cell Culture 336
Tumor Viruses Opened the Study of Cancer to
Molecular Methods 336
Retroviral Oncogenes Are Captured from
Cellular DNA 339
Viruses Can Activate Cellular Proto-
Oncogenes by Insertional Mutagenesis 340
Analysis of Tumor Chromosomes Reveals
Rearrangements of Proto-Oncogenes 340
The Transformed Phenotype Can Be Passed
via DNA-Mediated Transfection 341
An Activated Human Oncogene Is Cloned 341
The Human Bladder Carcinoma Oncogene Is
an Activated ras Gene 345
The Bladder Carcinoma ras Oncogene Is
Activated by Point Mutation 347
Proto-Oncogenes Encode Components of
Signal Transduction Pathways 349
Proto-Oncogenes Can Encode Growth Factors
or Their Receptors 349
Many Proto-Oncogenes Act as Intracellular
Signal Transducers 349
Some Proto-Oncogenes Are Transcription
Factors 351
Experimentally Modeling the Complexities of
Cancer Multiple Hits 352
Oncogenes Cause Cancer in Transgenic Mice 353
The Fusion Paradox: Normal Growth Is
Dominant to Transformation 355
Susceptibility to Cancer Can Be Inherited 355
The Cloning of the Retinoblastoma
Susceptibility Gene 357
The RB Protein Is the Target of DNA Tumor
Virus Oncogenes 358
The Strange Case of p53: An Oncogene
Crosses to the Other Side 358
The Several Steps to Colorectal Cancer
A Real-Life Tale of Oncogenes and
Antd-Oncogenes 361
Cancer Results from Accumulation of
Dominant and Recessive Mutations 363
19 MOLECULAR ANALYSIS OF
THE CELL CYCLE 369
There Are Four Stages in the Cell Cycle 370
There Are Two Types of Control of Cell
Division 370
Yeasts Are Good Systems for Studying Cell
Cycle Control 373
Temperature-Sensitive Mutants Are a Valuable
Tool in Studying the Cell Division Cycle 373
The CDC28 Gene Encodes a Ubiquitous
Protein Kinase 375
The Protein Kinase Activity of the Cdc2
Protein Varies with the Cell Cycle 376
MPF Contains Cdc2 377
Cyclins Are Cloned 380
Cyclin and Cdc2 Form a Protein Complex 380
Cyclin Destruction Is Required to Inactivate
the Kinase Activity 382
A Different Set of Cyclins Regulates Cdc28 in
Gl Phase 383
Other Cdc Proteins Are Also Involved in
Regulating Cdc2 Kinase Activity 386
Recombinant DNA Provides a Common
Language for Different Experimental
Systems 386
20 GENES THAT CONTROL THE
DEVELOPMENT OF DROSOPHILA 389
Serendipity and Systematic Genetic Screens
Identify Genes That Control Development 390
The First Drosopbila Development Genes Were
Cloned by Chromosome Walking 392
The Homeodomain Is Shared among Drosopbila
Developmental Genes 393
Anterior-Posterior Polarity Arises from a
Gradient of a Homeodomain Protein 394
GapGenesDefinetheFirstSubdivisionsofthe
Embryo 395
Segmentation Genes Divide the Embryo into
Stripes 395
The bicoid Protein Directly Regulates the
Transcription of hunchback 396
Expression in Stripes Is Encoded by Discrete
Cis-Acting Regulatory Elements 398
The Specificity of Homeotic Genes Remains
Unexplained 400
Formation of the Embryo s Ends Requires a
Protein Kinase-Dependent Signal
Transduction Pathway 401
Dorsal-Ventral Polarity Is Achieved by
Regulated Transport of a Transcription
Factor to the Nucleus 402
Drosopbila Developmental Genes Help Isolate
New Vertebrate Developmental Genes 404
21 THE GENES BEHIND THE
FUNCTIONING OF THE BRAIN 409
Genes Specifying the Development of Neurons
Are Cloned 410
XCONTENTS
Neurotrophic Factors Stimulate Neuron
Growth and Differentiation 412
Retroviruses Are Used to Mark and
Immortalize Neurons 413
Cloning and Mutagenesis Establish Structural
Models of the Voltage-Gated Ion Channels 416
Neurotransmitter Receptors Are Members of
Large Gene Families 418
Homology Cloning Is Used to Isolate
Components of Signal Transduction
Pathways in Olfactory Neurons 422
Learning and Memory Require Stable Changes
in Neuron Function 423
Molecular Cloning and Gene Mapping Begin
to Explore Alzheimer s Disease 428
22 RECOMBINANT DNA AND EVOLUTION 433
Life May Have Originated in an RNA World 435
DNA—Why So Much? 438
Genes Can Be Turned On (and Off) by
Movable Elements 439
Introns Are Ancient Components of Genes 440
Exon Shuffling Contributes to Gene Evolution 441
Gene Duplication Is a Driving Force in
Evolution 443
DNA Clocks Measure Rates of Evolution 444
rRNA Sequencing Identifies the Three Great
Kingdoms of Living Organisms 445
Recombinant DNA Techniques Have Sorted
Out Relationships in the Primate Family 445
Mitochondrial DNA as a Molecular Clock 446
Some Intracellular Organelles Were Once
Bacteria 447
DNA Fingerprinting Helps Us Understand the
Genetic Basis of Altruism 448
Application of Recombinant DNA
to Biotechnology
23 RECOMBINANT DNA IN MEDICINE
AND INDUSTRY 453
Expression Systems Are Developed to Produce
Recombinant Proteins 454
Insulin Is the First Recombinant Drug
Licensed for Human Use 455
Recombinant Human Growth Hormone Is
Produced in Bacteria by Two Methods 455
A Hepatitis B Virus Vaccine Is Produced in
Yeast by Expression of a Viral Surface
Antigen 458
Complex Human Proteins Are Produced by
Large-Scale Mammalian Cell Culture 458
Monoclonal Antibodies Function as Magic
Bullets 460
Human Antibodies That Recognize Specific
Antigens Can Be Directly Cloned and
Selected 461
Humanized Monoclonal Antibodies Retain
Activity But Lose Immunogenicity 464
Protein Engineering Can Tailor Antibodies for
Specific Applications 465
Protein Engineering Is Used to Improve a
Detergent Enzyme 465
Growth Hormone Variants with Improved
Binding Are Selected by Phage Display 466
New Technologies Promise New Approaches
to Drug Design 468 •
24 GENERATION OF AGRICULTURALLY
IMPORTANT PLANTS AND ANIMALS 471
Plants Expressing a Viral Coat Protein Resist
Infection 472
Insects Fail to Prey on Plants Expressing a
Bacterial Toxin 473
Herbicide-Tolerant Transgenic Plants Allow
More Effective Management of Weeds 473
Flowers Exhibiting New Colors and Patterns
Can Be Obtained by Genetic Engineering 475
The Potential Use of Plants to Produce
Proteins Is of Commercial Importance 477
Recombinant Bovine Growth Hormone
Stimulates Milk Production and Improves
Feed Utilization 478
Procedures for Generation of Transgenic Farm
Animals Are Developed 478
Pharmaceutical Proteins Can Be Produced in
Transgenic Animals 479
Farm Animals May Be Protected from Viral
Infection by Transgenic Expression of
Viral Coat Protein 481
The Implementation of Agricultural
Biotechnology Requires Continued
Research and Social Discussion 481
25 MARSHALING RECOMBINANT DNA
TO FIGHT AIDS 485
Human Immunodeficiency Virus Is the Cause
of AIDS 486
HIV Is a Retrovirus 486
HIV Belongs to the Most Complex Class of
Retroviruses Yet Discovered 488
The tat Gene Regulates Synthesis of HIV RNA
Movement of HIV RNA from Nucleus to
Cytoplasm Is Regulated by Rev
The Gene May Be Essential for in Vivo
Replication of Pathogenic AIDS
AZT Works by Interfering with Viral DNA
Synthesis
HIV Protease Is a Target for AIDS Drug
Therapy
HIV Infects and Kills T Lymphocytes That
Have the CD4 Receptor
Soluble CD4 Molecules Can Be Used to
Prevent HIV Infection
CD4-Toxin Conjugates Specifically Kill
HIV-infected Cells
Transport of HIV Proteins to the Cell Surface
Can Be Inhibited
Simian Immunodeficiency Virus and Animal
Models Are Useful in Studying AIDS
Recombinant HIV Proteins May Be Effective
as Immunogens for AIDS Vaccines
Kaposi s Sarcoma Is a Tumor Associated with
AIDS
The Origins and Evolution of the Human
Immunodeficiency Viruses Are Revealed
Through Recombinant Techniques
Recombinant DNA Is at the Forefront of the
Battle Against AIDS
Impact of Recombinant DNA on
Human Genetics
26 MAPPING AND CLONING HUMAN
DISEASE GENES
Human Genetic Diseases Have Simple and
Complex Patterns of Inheritance
The Metabolic Basis Is Known for Some
Human Inherited Diseases
Positional Cloning Uses the Location of a
Gene on a Chromosome to Clone the
Gene
Subchromosomal Mapping of Genes and
Markers Can Be Accomplished with
Somatic Cell Hybrids
Cloned Genes and Markers Can Be Localized
by in Situ Hybridization to Chromosomes
Chromosomal Abnormalities Provide Another
Means of Locating Disease Genes
Restriction Fragment Length Polymorphisms
Serve as Markers for Linkage Analysis
Contents xi
489 Linkage is Calculated from Frequency of
Recombination 522
491 Abnormal X Chromosomes Provide a Means of
Cloning the Gene for Duchenne Muscular
493 Dystrophy
cDNAs for the DMD Gene Were Cloned
494 Using Two Strategies
The Cystic Fibrosis Gene Was Cloned Using
496 RFLP Analysis and Chromosome Jumping
The Cystic Fibrosis Gene Was Identified by
496 DNA Sequencing
Clues about Protein Function Come from the
498 Sequences of Cloned Disease Genes
Cloned Genes Are Used to Study Protein
498 Expression and Function
Cloning Genes for Polygenic and Multifactorial
500 Disorders Is Difficult
Candidate Genes Can Be Used to Clone
500 Human Disease Genes
Cloning Human Disease Genes Will Continue
501 to Depend on Linkage Studies 533
504 27 DNA-BASED DIAGNOSIS OF
GENETIC DISEASES 539
504 Biochemical Markers Used for Early Diagnosis 540 Mutations in Globin Genes Cause the
505 Thalassemias 540 RFLPs and Linkage Analysis Are Used for
Diagnosis 545
Linkage Disequilibrium Can Be Used for
Diagnosis 547
Exon Deletions Are Used for Direct Diagnosis
of DMD 548
Gene Mutations Can Alter a Restriction
Enzyme Site: Direct Diagnosis of
511 Sickle-Cell Anemia 549 Allele-Specific Oligonucleotide Probes Are
Used to Detect Mutations 550
512 The Ligase-Mediated Technique Detects
Mutations 552
513 The Polymerase Chain Reaction
Revolutionizes DNA-Based Diagnosis 552
DNA Diagnostic Methods Are Used to
514 Distinguish Tumor Types
Novel Methods Are Developed for Screening
for Mutations 557
514 Genetic Testing May Bring Problems As Well
As Benefits 559
517 Hypervariable or Variable Tandem Repeat
Loci Can Be Used to Identify Individuals 561
517 DNA Fingerprinting Is Used in the Courts
Genetic Privacy Will Become an Important
519 Issue 563
xii CONTENTS
28 WORKING TOWARD HUMAN GENE
THERAPY
Gene Defects Have Been Corrected in
Transgenic Animals
Gene Therapy in Human Beings Raises Ethical
Issues
The Cystic Fibrosis Defect Can Be Corrected
in Vitro
Hematopoietic Cells Are Used for Expression
of Human Genes in Animals
Genetically Engineered Bone Marrow Cells
Survive for Long Periods in Vivo
Skin Fibroblasts Are Target Cells for Gene
Therapy
Hepatocytes May be Used for Gene Therapy
Gene Therapy Experiments Have Been
Conducted in Large Animals
Myoblast Transfer Is Used to Treat Duchenne
Muscular Dystrophy
Genes Can Be Delivered Directly to Target
Sites in Vivo
Genetically Modified Lymphocytes Have Been
Administered to Human Beings
Human Adenosine Deaminase Deficiency is
Treated by Gene Therapy
29 STUDYING WHOLE GENOMES
Very Large Pieces of DNA Can Be Separated
by Pulsed Eield Gel Electrophoresis
(PFGE)
PFGE Is Used to Make Large-Scale Physical
Maps
Putting Together the Cloned Genome of E coli
Requires Finding Overlapping Segments
Yeast Artificial Chromosomes (YACs) Are Used
for Cloning Huge DNA Fragments
YACs Are Used to Link the Cosmid Contigs of
C elegans
Cosmid and YAC Clones Are Ordered along
the Chromosomes of Drosopbila Salivary
Gland Cells by in Situ Hybridization
An Entire Yeast Chromosome Has Been
Sequenced 595
A Multiplex Method Speeds Up DNA
Sequencing 595
Automated DNA Sequencing Greatly Speeds
Up the Process 595
Understanding of DNA Sequences Is Furthered
by Homology Comparisons 598
Novel Methods Will Be Required for
Large-Scale Sequencing of DNA 600
30 THE HUMAN GENOME INITIATIVE-
FINDING ALL THE HUMAN GENES 603
Making a High-Resolution Genetic Map of
Humans Uses Reference Markers 605
Human Chromosomes Are Separated from
Each Other Using Cell Sorting Machines 606
DNA for Cloning Can Be Microdissected from
Human Chromosomes 606
Somatic Cell Hybrids Serve as Sources for
Purified Human Chromosome DNA 608
X-Irradiated Fragments of Human
Chromosomes Are Used for Gene
Mapping 608
Cloned Human DNA Fragments Must Be
Assembled into Megabase-Sized Contigs 609
Sequence-Tagged Sites Identify Cloned DNA 610
Complete Human Disease Genes Can Be
Reassembled in YACs 612
YACs Are Used to Clone the Telomeres of
Human Chromosomes 612
YACs Helped Unravel the Mysteries of the
Fragile X Region 613
Large-Scale Sequencing of the Human HPRT
Gene Was Done in Four Stages 614
Storing and Analyzing Genome Data Require
Large Databases 614
Gene Mapping Can Be Facilitated by
Comparing Species 615
Understanding Our Genome Will Benefit
Humanity 616
Index 619
|
| any_adam_object | 1 |
| building | Verbundindex |
| bvnumber | BV027261651 |
| classification_rvk | WD 5360 WG 3450 |
| ctrlnum | (OCoLC)299952225 (DE-599)BVBBV027261651 |
| discipline | Biologie |
| edition | 2. ed. |
| format | Book |
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| id | DE-604.BV027261651 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T22:56:58Z |
| institution | BVB |
| isbn | 0716722828 |
| language | Undetermined |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-021302058 |
| oclc_num | 299952225 |
| open_access_boolean | |
| owner | DE-188 DE-B768 |
| owner_facet | DE-188 DE-B768 |
| physical | XII, 626 S. Ill., graph. Darst. |
| publishDate | 1992 |
| publishDateSearch | 1992 |
| publishDateSort | 1992 |
| publisher | Freeman |
| record_format | marc |
| series2 | Scientific American books |
| spelling | Recombinant DNA James D. Watson ... 2. ed. New York Freeman 1992 XII, 626 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Scientific American books Recombinant DNA cabt Rekombinante DNS (DE-588)4225578-8 gnd rswk-swf Rekombination (DE-588)4049338-6 gnd rswk-swf DNS (DE-588)4070512-2 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Genetik (DE-588)4071711-2 gnd rswk-swf Rekombination (DE-588)4049338-6 s Genetik (DE-588)4071711-2 s Methode (DE-588)4038971-6 s DE-604 DNS (DE-588)4070512-2 s Rekombinante DNS (DE-588)4225578-8 s Watson, James D. Sonstige oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021302058&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Recombinant DNA Recombinant DNA cabt Rekombinante DNS (DE-588)4225578-8 gnd Rekombination (DE-588)4049338-6 gnd DNS (DE-588)4070512-2 gnd Methode (DE-588)4038971-6 gnd Genetik (DE-588)4071711-2 gnd |
| subject_GND | (DE-588)4225578-8 (DE-588)4049338-6 (DE-588)4070512-2 (DE-588)4038971-6 (DE-588)4071711-2 |
| title | Recombinant DNA |
| title_auth | Recombinant DNA |
| title_exact_search | Recombinant DNA |
| title_full | Recombinant DNA James D. Watson ... |
| title_fullStr | Recombinant DNA James D. Watson ... |
| title_full_unstemmed | Recombinant DNA James D. Watson ... |
| title_short | Recombinant DNA |
| title_sort | recombinant dna |
| topic | Recombinant DNA cabt Rekombinante DNS (DE-588)4225578-8 gnd Rekombination (DE-588)4049338-6 gnd DNS (DE-588)4070512-2 gnd Methode (DE-588)4038971-6 gnd Genetik (DE-588)4071711-2 gnd |
| topic_facet | Recombinant DNA Rekombinante DNS Rekombination DNS Methode Genetik |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021302058&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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