Drosophila: a practical approach
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Oxford [u.a.]
IRL Press at Oxford Univ. Press
1998
|
| Edition: | 2. ed. |
| Series: | The practical approach series
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XXIV, 389 S. Ill. |
| ISBN: | 0199636613 0199636605 |
Staff View
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Record in the Search Index
| _version_ | 1804126823841267713 |
|---|---|
| adam_text | Drosophila
A Practical Approach
Second Edition
Edited by
D B ROBERTS
Genetics Unit, Biochemistry Department, South Parks Road,
Oxford OXl 3QU, and Magdalen College Oxford OXl 4AU
-at-OXFORD UNIVERSITY PRESS
Oxford New York Tokyo
Contents
List of Con trib u tors xxi
Abbreviations xxin
1 The elements of Drosophila biology and
genetics 1
David B Roberts and Graeme N Standen
1 Introduction 1
2 Drosophila melanogaster 1
Classification 1
Distribution 2
Sibling species 2
3 Books 2
4 FlyBase and the Encyclopaedia of Drosophila 3
5 Drosophila stocks 5
Wild-type stocks 5
Mutant stocks 5
6 Chromosome aberrations 6
Polytene chromosome band nomenclature 7
Inversions 7
Deletions (deficiencies) 13
Duplications 17
Translocations 17
Compound chromosomes 18
7 Environmental conditions 19
Temperature 19
Humidity 20
8 Maintaining stocks 20
Temperature-sensitive (ts) stocks 20
9 Food 20
Flour-treacle-agar medium 20
Yeast-glucose-agar medium 23
Instant medium 24
Instant potato medium 24
Defined medium 25
Apple juice medium 27
Axenic medium 28
Contents
10 Pests 28
Fungal control 28
Bacterial control 29
Mite control 29
11 Handling flies 30
Anaesthetizers 31
Transferring flies 32
Distinguishing the sexes 33
Collecting virgins 34
Setting-up a cross 34
12 Simple genetic mapping 35
Assigning a gene to a chromosome 35
Mapping a gene on a chromosome 37
13 Drosophila life cycle 39
Larval growth 39
Metamorphosis 39
Development times 39
14 Mass culture 40
Small cages 40
Large cages 41
15 Eggs 43
Egg collecting from small cages 43
Egg collecting from large cages 43
Dechorionating eggs 44
Collecting timed eggs 45
16 Larvae 45
Collecting aged larvae 45
Harvesting larvae 46
Staging larvae 46
Recognizing chromosomes in larvae 48
Sexing larvae 48
17 Pupae 49
Definition of pupation 49
Staging pupae 49
Harvesting pupae 50
Collecting aged pupae 50
Recognizing chromosomes in pupae 50
18 Collecting flies in the wild 50
Identification of Drosophila melanogaster 51
19 Preparation of tissues 51
Fat body purification 52
Imaginal disc purification 53
Purification of other tissues 53
Acknowledgements 53
References 53
xii
Contents
2 Mutagenesis 55
T A Grigliatti
1 Introduction 55
2 Mutagens and methods of application 56
Ethyl methanesulfonate 56
N-ethyl-N-nitrosourea (ENU) 60
Triethylenemelamine (TEM) 61
Formaldehyde 62
Ionizing radiation 64
3 Protocols for identifying different types of mutations 64
Sex-linked recessive lethal mutations 65
Autosomal recessive lethal mutations 66
Non-lethal mutations 70
Conditional mutations 71
4 Systems to generate virgin females 74
5 Protocol for isolating deletions of specific regions 77
Pseudodominance of recessive allele, or reversion of a neomorph or
antimorph 78
6 Duplications 81
References 83
3 Transposons—gene tagging and mutagenesis 85
T A Grigliatti
1 Introduction 85
2 Historical background—hybrid dysgenesis 86
P-M hybrid dysgenesis 87
3 Assays for hybrid dysgenesis 89
Gonadal dysgenesis—a morphological assay 89
snw—a genetic assay 89
4 Using P elements to produce novel mutations 89
P[ry+ (b2-3)] 92
Screening for genes by selecting for specific phenotypes—tagging
genes that are not characterized 92
Tagging a gene for which mutants are described and genetically
and/or cytogenetically mapped 94
Creating new alleles in a P tagged gene (including reversions) 98
Inducing deletions with denned end-points 99
5 Selection for P element movement to non-homologous
chromosome 101
References 106
xiii
Contents
4 Chromosome mechanics; the genetic
manipulation of aneuploid stocks 109
David Gubb
1 Introduction 109
2 The generation of aneuploids (either deletions or
duplications) from translocations by segregation 110
The T(Y;A) translocatioin set of Lindsley and Sandier et al 112
3 The generation of aneuploids by recombination 114
Transpositions 114
Inversions 115
4 Craymer s autosynaptic method 117
The recovery of aneuploid autosynaptic stocks 118
Construction of marked autosynaptic elements 120
The construction of synthetic aneuploid using h;A autosynpatic stocks 121
Conversion of T(Y;A) translocations to autosynaptic form 122
Resolution of autosynaptic chromosomes 124
5 Selective genetic screens 127
Second generation selective screens 127
6 The use of aneuploid stocks to identify enhancers and
suppressors 128
The CAM duplication set 129
Acknowledgements 130
References 130
5 Enhancer traps 131
C J O Kane
1 Enhancer trapping—principles and uses 131
What are enhancer traps? 131
Enhancer traps as cell markers 132
Targeted expression with GAM 132
Gene discovery using enhancer traps 134
2 Generating new enhancer trap insertions 135
Target specificity 135
Choices of P-lacZ vector 136
Generating new insertions 137
3 LacZ enhancer traps as cell markers 142
4 Manipulating cells with P-GAL4 enhancer traps 150
Components of the system 150
Practical considerations 151
Altering and disabling cells 154
xiv
Contents
5 Alternative reporter genes
LacZ nuclear and cytoplasmic localization
Visualizing cellular structure
In vivo reporting—green fluorescent protein
Other in vivo reporter genes
Epitope tags
6 You ve trapped your enhancer—can you trap your gene?
Initial steps
Enhancer trap insertions and Drosophila genome maps
Plasmid rescue
Detecting flanking transcripts
7 Generating a mutant
Has the insertion caused a mutation?
Which gene is affected? Is the mutation a null?
Generating (more) mutant alleles by imprecise excision
Which gene is responsible for the mutant phenotype?
8 An enhancer trap miscellany
Modular misexpression
Enhancer piracy
Stripe traps
Enhancer trap targeting
Acknowledgements
References
6 Looking at embryos 179
Eric Wieschaus and Christiane Nusslein-Volhard
1 Introduction 179
2 Collecting embryos 179
Care of adults 180
Collecting large numbers of embryos from a single stock 180
Collecting eggs from multiple stocks 181
Collecting eggs from single females 182
Preparation of agar plates 182
Collection intervals and maintaining plates 183
3 Looking at l iving embryos 183
Scoring late stages directly on plates under a stereomicroscope 183
Scoring early stages on plates under stereomicroscope; determining
earliest phenotypic manifestations 184
Photographing single embryos using the compound microscope 184
Videotaping living embryos 185
Klarsicht 186
4 Embryonic stages and morphological criteria for identifying
approximate ages of living embryos 186
xv
Contents
Stage 1: freshly laid egg 186
Stage 2: early cleavage 186
Stage 3: pole cell formation 187
Stage 4: syncytial blastoderm 188
Stage 5: cell formation 188
Stage 6: early gastrulation, ventral furrow formation 188
Stage 7: midgut invaginations 188
Stage 8: germ band extension 190
Stage 9: stomodeal plate formation 192
Stage 10: stomodeal invagination 194
Stage 11: three-layered germ band 196
Stage 12: shortening of germ band 196
Stage 13: shortened embryo 196
Stage 14: head involution and dorsal closure 196
Stage 15: dorsal closure complete 197
Stage 16: condensation of CNS 197
5 Cuticle preparations 197
Embryo preparations 197
Other tricks for especially flat preparations of high contrast 200
Analysis of cuticle patterns 201
Photographing cuticle preparations 201
6 Whole mount procedure for looking at embryos at early stages 201
Fixation of embryos 201
Heat methanol fixation 205
Staining and analysis of whole mounts 206
Protocols for scanning electron microscopy 211
References 213
7 Immunolabelling of Drosophila 215
Robert A H White
1 Introduction 215
2 Antibodies 215
Types of antibody 215
Storage 216
Antibody specificity 217
Antibody binding 217
Fixation 218
Detection 218
3 Embryo immunolabelling 219
Embryo fixation 219
Labelling of intact embryos 220
Labelling of flat preparations or filets 224
4 Immunolabelling of imaginal discs 228
xvi
Contents
5 Immunolabelling of ovaries 229
6 Double and multiple labelling 231
7 Immunolabelling of salivary glands 234
8 Immunolabelling of sections 237
9 Vexations and variations 237
Background labelling problems 237
Weak labelling 238
Acknowledgements 239
References 239
8 Population and ecological genetics 241
J F YBrookfield
1 Introduction
2 Experimental methods
Maintenance of the genetic variability of wild populations of
D melanogaster in the laboratory
Quantitative genetics
Enzyme electrophoresis
Measurement of fitness
Measurement of DNA sequence variation
3 Mathematical and statistical analyses
Hardy-Weinberg equilibrium and estimation of allele frequencies
Population subdivision
Linkage disequilibrium
Estimation of evolutionary distances and rates
Phylogeny estimation
Estimates of DNA sequence diversity
Tests for natural selection
4 Discussion
References
9 Behaviour, learning, and memory 265
John B Connolly and Tim Tully
1 Introduction 265
Behaviour and behavioural plasticity 265
Habituation, sensitization, and adaptation 266
Associative learning 266
2 Maintenance and preparation of stocks 267
Genetic background 267
xvii
Contents
Food, medication, and age 268
Preparation of flies 271
Preparation of larvae 272
3 Locomotion 272
Larval locomotor activity 272
Fly locomotor activity 272
4 Mechanosensation 275
Larval mechanosensation 275
Adult grooming 276
Habituation of the cleaning reflex 277
5 Chemosensation 278
Larval chemosensation 278
The proboscis extension reflex 278
6 Olfaction 281
Larval olfaction 281
The olfactory trap assay 281
The olfactory jump response 282
Habituation of the olfactory jump response 283
Behaviour in the T -maze 284
7 Associative olfactory learning and memory 287
Olfactory learning and memory in larvae 287
The teaching machine 288
Teaching machine preparation and maintenance 289
Preparation of collection and cover tubes 290
Preparation of odour cups 291
Preparation of grid tubes 293
Vacuum source and flowmeters 294
Calculating the performance index for olfactory learning 296
Spaced and massed training 298
Peripheral behaviours 298
The appearance of memory 300
Dissecting the phases of memory 300
Mutations impairing olfactory learning and memory 301
Behavioural pharmacology 302
Transgenes affecting learning and memory 305
8 Vision 306
Larval phototaxis 307
Slow phototaxis in adults 308
Fast phototaxis in adults 308
The optomotor response 309
Plasticity of the visual response 310
The landing response and its habituation 310
The flight simulator 311
Visual operant conditioning 311
xviii
Contents
9 Courtship 312
Courtship 312
Courtship conditioning 312
Inhibition of courtship to immature males 315
Acoustic priming 315
10 Statistics 315
Acknowledgements 315
References 315
10 Cell culture 319
Lucy Cherbas and Peter Cherbas
1 Introduction 319
2 Cell lines 319
Embryonic lines 319
Lines from defined cell types 321
3 General guidelines 321
Sterility 321
Cleanliness 322
Water quality 323
4 Maintenance of cell lines 323
Media 323
Serum 324
Other additions to media, 325
Saline solutions 326
Culture procedures 326
5 Transfection 331
Procedures 331
Strategies 335
Acknowledgements 336
References 336
Appendix 1 339
11 Preparation of nucleic acids 347
T Jowett
1 Introduction 347
General precautions 347
Safety considerations 349
Assaying yields and quality 349
xix
Contents
2 DNA extraction procedures 350
Preparation of high molecular weight genomic DNA 350
» Medium scale preparation of genomic DNA 353
Single fly extraction of DNA 355
3 RNA extraction procedures 357
Extraction of total RNA 359
Simultaneous isolation of DNA and RNA 362
Direct methods of purification of mRNA 363
4 Purification of mitochondrial DNA 367
References 370
List of suppliers 373
Index o/Drosophila genes, mutations,
chromosal abnormalities and balancers 379
General index 385
XX
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| discipline | Biologie |
| edition | 2. ed. |
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| genre | 1\p (DE-588)4143413-4 Aufsatzsammlung gnd-content |
| genre_facet | Aufsatzsammlung |
| id | DE-604.BV012205816 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T18:23:35Z |
| institution | BVB |
| isbn | 0199636613 0199636605 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008271658 |
| oclc_num | 37820098 |
| open_access_boolean | |
| owner | DE-20 DE-703 DE-355 DE-BY-UBR |
| owner_facet | DE-20 DE-703 DE-355 DE-BY-UBR |
| physical | XXIV, 389 S. Ill. |
| publishDate | 1998 |
| publishDateSearch | 1998 |
| publishDateSort | 1998 |
| publisher | IRL Press at Oxford Univ. Press |
| record_format | marc |
| series2 | The practical approach series |
| spelling | Drosophila a practical approach ed. by D. B. Roberts 2. ed. Oxford [u.a.] IRL Press at Oxford Univ. Press 1998 XXIV, 389 S. Ill. txt rdacontent n rdamedia nc rdacarrier The practical approach series Drosophila Drosophila Genetics Drosophila genetics Insects Genetics Praktikum (DE-588)4127380-1 gnd rswk-swf Drosophila (DE-588)4150708-3 gnd rswk-swf Molekulargenetik (DE-588)4039987-4 gnd rswk-swf Fruchtfliegen (DE-588)4071480-9 gnd rswk-swf Taufliege (DE-588)4078168-9 gnd rswk-swf Genetik (DE-588)4071711-2 gnd rswk-swf 1\p (DE-588)4143413-4 Aufsatzsammlung gnd-content Taufliege (DE-588)4078168-9 s Molekulargenetik (DE-588)4039987-4 s DE-604 Drosophila (DE-588)4150708-3 s Praktikum (DE-588)4127380-1 s 2\p DE-604 Genetik (DE-588)4071711-2 s 3\p DE-604 4\p DE-604 Fruchtfliegen (DE-588)4071480-9 s 5\p DE-604 Roberts, David B. Sonstige oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008271658&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 2\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 3\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 4\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 5\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Drosophila a practical approach Drosophila Drosophila Genetics Drosophila genetics Insects Genetics Praktikum (DE-588)4127380-1 gnd Drosophila (DE-588)4150708-3 gnd Molekulargenetik (DE-588)4039987-4 gnd Fruchtfliegen (DE-588)4071480-9 gnd Taufliege (DE-588)4078168-9 gnd Genetik (DE-588)4071711-2 gnd |
| subject_GND | (DE-588)4127380-1 (DE-588)4150708-3 (DE-588)4039987-4 (DE-588)4071480-9 (DE-588)4078168-9 (DE-588)4071711-2 (DE-588)4143413-4 |
| title | Drosophila a practical approach |
| title_auth | Drosophila a practical approach |
| title_exact_search | Drosophila a practical approach |
| title_full | Drosophila a practical approach ed. by D. B. Roberts |
| title_fullStr | Drosophila a practical approach ed. by D. B. Roberts |
| title_full_unstemmed | Drosophila a practical approach ed. by D. B. Roberts |
| title_short | Drosophila |
| title_sort | drosophila a practical approach |
| title_sub | a practical approach |
| topic | Drosophila Drosophila Genetics Drosophila genetics Insects Genetics Praktikum (DE-588)4127380-1 gnd Drosophila (DE-588)4150708-3 gnd Molekulargenetik (DE-588)4039987-4 gnd Fruchtfliegen (DE-588)4071480-9 gnd Taufliege (DE-588)4078168-9 gnd Genetik (DE-588)4071711-2 gnd |
| topic_facet | Drosophila Drosophila Genetics Drosophila genetics Insects Genetics Praktikum Molekulargenetik Fruchtfliegen Taufliege Genetik Aufsatzsammlung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008271658&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT robertsdavidb drosophilaapracticalapproach |