Fly pushing : the theory and practice of Drosophila genetics / Ralph J. Greenspan.

Format:

Book

Author/Creator:

Greenspan, Ralph J.

Language:

English

Subjects (All):

Drosophila--Genetics.

Drosophila.

Drosophila--genetics.

Medical Subjects:

Drosophila--genetics.

Physical Description:

xiv, 191 pages : illustrations ; 23 cm

Edition:

Second edition.

Place of Publication:

Cold Spring Harbor, N.Y. : Cold Spring Harbor Laboratory Press, [2004]

Contents:

Frequently Used Markers xiv

Chapter 1 The Basics of Doing a Cross 3

Fly Chromosomes 3

Recognizing Markers 6

Evolution of Gene Names 13

What Makes Flies So Great? Balancer Chromosomes 13

Deciphering Mating Schemes 15

Basic Fly Husbandry 18

Coddling Difficult Strains 20

Collecting Flies for Crosses 21

Chapter 2 Isolating New Variants 25

Principles 25

Mutagens 27

Ethylmethanesulfonate 27

Ethylnitrosourea 28

Radiation 29

Insertional Mutagenesis 29

Genetic Schemes for Detecting Mutants 31

New Alleles at an Autosomal Locus 31

New Alleles on the X 33

Identifying New Genes by Mutagenesis 36

Isogenizing the Starter Stock 36

The Mutagenesis Screen 37

Screening the X for New Genes 40

Insertional Mutagenesis and Enhancer Trapping 41

Enhancer-trap Screens 42

New Insertional Alleles of Known Loci 45

Mutating Genes for Which There Are No Allelic Differences 45

Screening Over a Deletion of the Locus 45

Targeted Insertional Mutagenesis 46

Homologous Recombination 46

RNA Interference 50

Searching for Gain of Function 50

Generating Mutants: One Size Does Not Fit All 51

Chapter 3 Mapping 53

Segregation Analysis 54

Problems with Balancers 55

Meiotic Mapping 57

Deletion Mapping 61

Duplication Mapping 62

Synthesizing Deletions and Duplications 63

Identifying New Embryonic Genes by Deletion Mapping 66

Mapping P-Element Insertion Sites 69

Fine-Grain Traditional Meiotic Mapping with P-Element Markers 69

Mapping by P-Element-Induced Male Recombination 71

Fine-Grain Mapping by Transposon-Induced Deletions 73

Mapping by DNA Polymorphisms 74

Chapter 4 Synthesizing Specific Genotypes 77

Principles 77

Simple Manipulations of a Single Chromosome 78

Linking Mutations 78

Manipulating Two Chromosomes 81

Pitfalls 85

How Many Flies? 85

Which Sex? 85

Manipulating Three Chromosomes 87

Making Rearrangements 88

Inducing Deletions 89

Synthesizing Stable Deletions from T(Y;A)s 92

Synthesizing Deletions from P-element Insertions 93

Inducing Duplications 95

Synthesizing Stable Duplications from T(Y;A)s 96

Synthesizing Attached-Xs 98

Autosynaptic Chromosomes and the Joys of Gibberish 99

Chapter 5 Analysis of Mutations: I. Characteristics of the Allele 101

Formal Classification of Mutations 101

Principles 101

Null Alleles (Amorphs) 102

Hypomorphs 103

Hypermorphs 106

Neomorphs 106

Antimorphs 107

Gain Versus Loss of Function 108

Intragenic Interactions 108

Interacting Loci 109

Ordering Genes in a Pathway 110

Dosage-dependent Interactions 112

Conditional Alleles 113

Inducible Promoters 115

Maintenance of Mutant Phenotypes 117

Genetic Background 118

Chapter 6 Analysis of Mutations: II. Mosaics 121

Principles 121

Chromosome Loss 122

Uses of Chromosome Loss Mosaics 123

Fate Mapping 123

Focus of Gene Action 124

Methods for Inducing Chromosome Loss 125

Ring Loss 125

Chromosome Loss Mosaics for Autosomal Genes 127

paternal-loss (pal) 128

claret-non-disjunctional (cand) 129

Care of Mosaic-producing Stocks 130

Mitotic Recombination 130

Synchronizing Age of Cultures 133

Uses of Mitotic Recombination Mosaics 134

Lineage Analysis 134

Focus of Gene Action 135

Time of Gene Action 136

Methods for Producing Mitotic Recombinant Clones 138

Markers for Internal Tissues 140

GAL4 and Expression Mosaicism 140

Cell Ablations 142

GAL4 Targeted FLP/FRT Recombination 142

Isolating Tissue-targeted Mutations Using Mitotic Recombination 144

P-element Replacement for Exploitation of Existing Enhancer Traps 145

Chapter 7 Commencement 147.

Notes:

Includes bibliographical references (pages 171-182) and index.

ISBN:

0879697113

OCLC:

54400228