Genetic Theory and Analysis
John Wiley & Sons Inc (Verlag)
978-1-118-08692-6 (ISBN)
Understand and apply what drives change of characteristic genetic traits and heredity
Genetics is the study of how traits are passed from parents to their offspring and how the variation in those traits affects the development and health of the organism. Investigating how these traits affect the organism involves a diverse set of approaches and tools, including genetic screens, DNA and RNA sequencing, mapping, and methods to understand the structure and function of proteins. Thus, there is a need for a textbook that provides a broad overview of these methods.
»Genetic Theory and Analysis« meets this need by describing key approaches and methods in genetic analysis through a historical lens. Focusing on the five basic principles underlying the field — mutation, complementation, recombination, segregation, and regulation — it identifies the full suite of tests and methodologies available to the geneticist in an age of flourishing genetic and genomic research.
This second edition of the text has been updated to reflect recent advances and increase accessibility to advanced undergraduate students.
»Genetic Theory and Analysis«, 2nd edition readers will also find:
- Detailed treatment of subjects including mutagenesis, meiosis, complementation, suppression, and more
- Updated discussion of epistasis, mosaic analysis, RNAi, genome sequencing, and more
- Appendices discussing model organisms, genetic fine-structure analysis, and tetrad analysis
»Genetic Theory and Analysis« is ideal for both graduate students and advanced undergraduates undertaking courses in genetics, genetic engineering, and computational biology.
Danny E. Miller, MD, PhD is an Assistant Professor in the Department of Pediatrics, Division of Genetic Medicine and Laboratory Medicine & Pathology at the University of Washington in Seattle, WA, USA. He is the recipient of the 2017 Larry Sandler Memorial Award, the 2018 Lawrence E. Lamb Prize for Medical Research, and a 2022 National Institutes of Health Director’s Early Independence Award. Dr Miller is a leader in the field of long-read sequencing technology and the use of new technology to evaluate individuals with unsolved genetic disorders.
Angela L. Miller is a Research Coordinator at the University of Washington in Seattle, WA, USA, with a background in journalism, visual communications, and molecular biology. She has published several peer-reviewed papers and has won multiple national awards for her work as a journal art director.
R. Scott Hawley, PhD is an Investigator at the Stowers Institute for Medical Research, Kansas City, MO, USA. He is a member of the National Academy of Sciences and former President of the Genetics Society of America, with faculty positions at the University of Kansas Medical Center and the University of Missouri-Kansas City. During his distinguished career, Dr. Hawley has mentored hundreds of trainees, received numerous genetics awards, written six textbooks, and published extensively on meiosis.
Preface
Introduction
Chapter 1: Mutation
This chapter describes different types of mutations and the various terminology used to describe mutations.
1.1 Types of Mutations
Muller’s classification of mutants
– Nullomorphs
– Hypomorphs
– Hypermorphs
– Antimorphs
– Neomorphs
Modern mutant terminology
– Loss-of-function mutants
– Dominant mutants
– Gain-of-function mutants
– Separation-of-function mutants
DNA-level terminology
– Base-pair-substitution mutants
– Base-pair insertions or deletions
– Chromosomal aberrations
1.2 Dominance and recessivity
The cellular meaning of dominance
The cellular meaning of recessivity
Difficulties in applying the terms dominant and recessive to sex-linked mutants
The genetic utility of dominant and recessive mutants
Summary
Boxes
Box 1.1 DNA-level terminology
Box 1.2 Detecting gene expression by RNA-seq
Box 1.3 De novo mutation
References
Chapter 2: Mutant Hunts
This chapter describes why identifying new genetic mutants is useful, ways to create mutants, and how to screen for mutant phenotypes.
2.1 Why look for new mutants?
Reason 1: To identify genes required for a specific biological process
Reason 2: To isolate more mutations in a specific gene of interest
Reason 3: To obtain mutants for a structure-function analysis
Reason 4: To isolate mutations in a gene so far identified only by computational approaches
2.2 Mutagenesis and mutational mechanisms
Method 1: Ionizing radiation
Method 2: Chemical mutagens
– Alkylating agents
– Crosslinking agents
Method 3: Transposons
– Identifying where your transposon landed
– Why not always screen with TEs?
Method 4: Targeted gene disruption
– RNA interference
– CRISPR/Cas9
– TALENs
So which mutagen should you use?
2.3 What phenotype should you screen (or select) for?
2.4 Actually getting started
Your starting material
Pilot screen
What to keep?
How many mutants is enough?
– Estimating the number of genes not represented by mutants in your new collection
Summary
Boxes
Box 2.1 A screen for embryonic lethal mutations in Drosophila
Box 2.2 A screen for sex-linked lethal mutations in Drosophila
– Objective
– Basic stocks
– The screen itself
– A complication
Box 2.3 The balancer chromosome
Box 2.4 De novo genome and transcriptome assembly
Box 2.5 Identifying new transposon insertion sites
References
Chapter 3: Complementation
This chapter describes methods for determining whether mutants isolated in a genetic screen are novel.
3.1 The essence of the complementation test
3.2 Rules for using the complementation test
The complementation test can be done only when both mutants are fully recessive
The complementation test does not require that the two mutants have exactly the same phenotype
There are cases where the phenotype of a compound heterozygote is more extreme than is that of either homozygote
3.3 How might the complementation test lie to you?
Two mutations in the same gene complement each other
A mutation in one gene silences expression of a nearby gene
Mutations in regulatory elements
3.4 Second-site noncomplementation (nonallelic noncomplementation)
Type 1 SSNC (poisonous interactions): the interaction is allele specific at both loci
– An example of type 1 SSNC involving the alpha- and beta-tubulin genes in yeast
– An example of type 1 SSNC involving the actin genes in yeast
Type 2 SSNC (sequestration): the interaction is allele specific at one locus
– An example of type 2 SSNC involving the tubulin genes in Drosophila
– An example of type 2 SSNC in Drosophila that does not involve the tubulin genes
– An example of type 2 SSNC in the nematode Caenorhabditis elegans
Type 3 SSNC (combined haploinsufficiency): the interaction is allele independent at both loci
– An example of type 3 SSNC involving two motor protein genes in flies
Summary of SSNC in model organisms
SSNC in humans (digenic inheritance)
Pushing the limits: third-site noncomplementation
3.5 An extension of SSNC: dominant enhancers
A successful screen for dominant enhancers
Summary
Boxes
Box 3.1 A more rigorous definition of the complementation test
Box 3.2 An example of using the complementation test in yeast
Box 3.3 Transformation rescue is a variant of the complementation test
Box 3.4 A method for determining whether a dominant mutation is an allele of a given gene
Box 3.5 Pairing-dependent complementation: transvection
Box 3.6 Synthetic lethality and genetic buffering
References
Chapter 4: Recombination
This chapter provides a description of meiotic recombination and how it is used to map the genomic regions affected by novel mutations.
4.1 An introduction to meiosis
A cytological description of meiosis
A more detailed description of meiotic prophase
4.2 Crossing over and chiasmata
4.3 The classical analysis of recombination
4.4 Measuring the frequency of recombination
The curious relationship between the frequency of recombination and chiasma frequency
Map lengths and recombination frequency
– The mapping function
Tetrad analysis
Statistical estimation of recombination frequencies
– Two-point linkage analysis
– What constitutes statistically significant evidence for linkage?
– An example of LOD score analysis
– Multipoint linkage analysis
– Local mapping via haplotype analysis
– The endgame
The actual distribution of exchange events
The centromere effect
The effects of heterozygosity for aberration breakpoints on recombination
Practicalities of mapping
4.5 The mechanism of recombination
Gene conversion
Early models of recombination
– The Holliday model
– The Meselson-Radding model
The currently accepted mechanism of recombination: the double-strand break repair model
Class I versus class II recombination events
Summary
Boxes
Box 4.1 The molecular biology of synapsis
Box 4.2 Do specific chromosomal sites mediate pairing?
– The role of telomeres in early pairing
– The role of centric heterochromatin in chromosome pairing
– Specific pairing sites in C. elegans
– Specific euchromatic pairing sites in Drosophila
Box 4.3 Crossing over in compound-X chromosomes
Box 4.4 Does any sister-chromatid exchange occur during meiosis?
– Genetic studies in yeast
– Genetic studies in Drosophila
– A direct molecular assessment in yeast
References
Chapter 5: Finding Homologous Genes
This chapter describes methods for determining whether a gene of interest identified in one organism has been described in another organism.
5.1 Homology
Orthologs
Paralogs
Xenologs
5.2 Identifying sequence homology
Nucleotide–nucleotide BLAST (blastn)
– An example using blastn
Translated nucleotide–protein BLAST (blastx)
– An example using blastx
Protein–protein BLAST (blastp)
– An example using blastp
Translated BLASTx (tblastx) and translated BLASTn (tblastn)
5.3 How similar is similar?
Summary
References
Chapter 6: Suppression
This chapter discusses how a mutant might suppress the phenotype of another mutant and how to screen for such suppressor mutants.
6.1 Intragenic suppression
Intragenic suppression of loss-of-function mutations
– Intragenic suppression of a frameshift mutation by the addition of a second, compensatory frameshift mutation
– Intragenic suppression of missense mutations by the addition of a second and compensatory missense mutation
– Intragenic suppression of antimorphic mutations that produce a poisonous protein
6.2 Extragenic suppression
6.3 Transcriptional suppression
Suppression at the level of gene expression
A CRISPR screen for suppression of inhibitor resistance in melanoma
Suppression of transposon-insertion mutants by altering the control of mRNA processing
Suppression of nonsense mutants by messenger stabilization
6.4 Translational suppression
tRNA-mediated nonsense suppression
– The numerical and functional redundancy of tRNA genes allows suppressor mutations to be viable
tRNA-mediated frameshift suppression
6.5 Suppression by post-translational modification
6.6 Conformational suppression: suppression as a result of protein–protein interaction
Searching for suppressors that act by protein-protein interaction in eukaryotes
– Actin and fimbrin in yeast
– Mediator proteins and RNA polymerase II in yeast
“Lock-and-key” conformational suppression
– Suppression of a flagellar motor mutant in E. coli
– Suppression of a mutant transporter gene in C. elegans
– Suppression of a telomerase mutant in humans
6.7 Bypass suppression: suppression without physical interaction
“Push me, pull you” bypass suppression
Multicopy bypass suppression
6.8 Suppression of dominant mutations
6.9 Designing your own screen for suppressor mutations
Summary
Boxes
Box 6.1 Bypass suppression of a telomere defect in the yeast S. pombe
References
Chapter 7: Epistasis Analysis
This chapter describes methods for determining whether different genes function in the same biological pathway and, if so, the order in which they function.
7.1 Ordering gene function in pathways
Biosynthetic pathways
Nonbiosynthetic pathways
7.2 Dissection of regulatory hierarchies
Epistasis analysis using mutants with opposite effects on the phenotype
– Hierarchies for sex determination in Drosophila
Epistasis analysis using mutants with the same or similar effects on the final phenotype
– Using opposite-acting conditional mutants to order gene function by reciprocal shift experiments
– Using a drug or agent that stops the pathway at a given point
– Exploiting subtle phenotypic differences exhibited by mutants that affect the same signal state
7.3 How might an epistasis experiment mislead you?
Summary
References
Chapter 8: Mosaic Analysis
This chapter describes methods for determining in which tissue(s) or at what stage(s) of development a given gene functions.
8.1 Tissue transplantation
Early tissue transplantation in Drosophila
Tissue transplantation in zebrafish
8.2 Mitotic chromosome loss
Loss of the unstable ring X chromosome
Other mechanisms of mitotic chromosome loss
Mosaics derived from sex chromosome loss in humans and mice (Turner syndrome)
8.3 Mitotic recombination
Gene knockout using the FLP/FRT or Cre-Lox systems
8.4 Tissue-specific gene expression
Gene knockdown using RNAi
Tissue-specific gene editing using CRISPR/Cas9
Summary
Boxes
Box 8.1 The ethics of targeted gene editing in humans
References
Chapter 9: Meiotic Chromosome Segregation
This chapter describes the mechanisms that ensure meiotic chromosome segregation, which is the physical basis of Mendelian inheritance.
9.1 Types and consequences of failed segregation
9.2 The origin of spontaneous nondisjunction
MI exceptions
MII exceptions
9.3 The centromere
The isolation and analysis of the S. cerevisiae centromere
The isolation and analysis of the Drosophila centromere
The concept of the epigenetic centromere in Drosophila and humans
Holocentric chromosomes
9.4 Chromosome segregation mechanisms
Chiasmate chromosome segregation
Segregation without chiasmata (achiasmate chromosome segregation)
– Achiasmate segregation in Drosophila males
– Achiasmate segregation in Drosophila females
– Achiasmate segregation in S. cerevisiae
– Achiasmate segregation in S. pombe
– Achiasmate segregation in silkworm females
9.5 Meiotic drive
Meiotic drive via spore killing
– An example in Schizosaccharomyces pombe
– An example in Drosophila melanogaster
Meiotic drive via directed segregation
Summary
Boxes
Box 9.1 Identifying genes that encode centromere-binding proteins in yeast
Box 9.2 Achiasmate heterologous segregation in Drosophila females
Figures
References
Appendix A: Model Organisms
This appendix presents useful information for performing genetic analyses in the various model organisms mentioned throughout this book.
A.1 Budding yeast: Saccharomyces cerevisiae
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.2 Plants: Arabidopsis thaliana
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.3 Worms: Caenorhabditis elegans
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.4 Fruit flies: Drosophila melanogaster
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.5 Zebrafish: Danio rerio
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.6 Mice: Mus musculus
Basic culture techniques
Nomenclature
Chromosome biology
Useful guides and manuals
A.7 Phage lambda
Nomenclature
Useful guides and manuals
Appendix B: Genetic Fine-Structure Analysis
This appendix describes the classical approach to mapping the exact location of a mutation within a gene.
B.1 Intragenic mapping (then)
The first efforts toward finding structure within a gene
The unit of recombination and mutation is the base pair
B.2 Intragenic complementation meets intragenic recombination: the basis of fine-structure analysis
The formal analysis of intragenic complementation
B.3 Fine-structure analysis of a eukaryotic gene encoding a multifunctional protein
Genetic and functional dissection of the HIS4 gene in yeast
Genetic and functional dissection of the rudimentary gene in Drosophila
B.4 Fine-structure analysis of genes with complex regulatory elements in eukaryotes
Genetic and functional dissection of the cut gene in Drosophila
B.5 Pairing-dependent intragenic complementation
Genetic and functional dissection of the yellow gene in Drosophila
The influence of the zeste gene on pairing-dependent complementation at the white locus in Drosophila
Genetic and functional dissection of the bithorax complex in Drosophila
Summary
References
Appendix C: Tetrad Analysis
This appendix describes approaches for measuring map length and the frequency of recombination.
C.1 Tetrad analysis in linear asci
C.2 Unordered tetrad analysis
C.3 Half-tetrad analysis
C.4 Algebraic tetrad analysis
A simple example of algebraic tetrad analysis
A more complicated example of algebraic tetrad analysis
Boxes
Box C.1 Using tetrad analysis to determine linkage
Box C.2 Mapping centromeres in fungi with unordered tetrads
References
Glossary
Index
| Erscheinungsdatum | 19.06.2019 |
|---|---|
| Verlagsort | New York |
| Sprache | englisch |
| Maße | 150 x 250 mm |
| Gewicht | 612 g |
| Einbandart | kartoniert |
| Themenwelt | Medizin / Pharmazie ► Medizinische Fachgebiete ► Biomedizin |
| Medizin / Pharmazie ► Medizinische Fachgebiete ► Mikrobiologie / Infektologie / Reisemedizin | |
| Naturwissenschaften ► Biologie | |
| Technik ► Umwelttechnik / Biotechnologie | |
| ISBN-10 | 1-118-08692-9 / 1118086929 |
| ISBN-13 | 978-1-118-08692-6 / 9781118086926 |
| Zustand | Neuware |
| Haben Sie eine Frage zum Produkt? |
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