Tetrahymena thermophila:
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
San Diego [u.a.]
Acad. Press
2000
|
| Schriftenreihe: | Methods in cell biology
62 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVIII, 580 S. Ill., graph. Darst. |
| ISBN: | 0125441649 0120645254 |
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Datensatz im Suchindex
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|---|---|
| adam_text | CONTENTS
Contributors xiii
Preface xvii
PART I Overview
1. Laboratory and Evolutionary History of Tetrahytnena thermophila
David L. Nanney and Ellen M. Simon
Summary 4
I. Characters in Search of a Script 4
II. Paramecium Precursors 8
III. Tetrahymena Breeding Studies 11
IV. Perils of Passage 16
V. Doing Genetics by Evolution 18
References 22
2. Cell Biology of Tetrahymena thermophila
Joseph Frankel
I. Introduction 28
II. Membranes and Membrane Bounded Compartments 35
III. The Cytoskeleton and Membrane Skeleton 52
IV. Dynamics of the Cell Cycle 68
V. Spacial Patterning 79
VI. Starvation Mediated Development 88
VII. Concluding Remarks 99
References 103
3. Tetrahymena Genetics: Two Nuclei Are Better Than One
Kathleen M. Karrer
I. Asexual and Sexual Reproduction 128
II. Nuclear Dualism 134
III. Special Features of Tetrahymena Genetics 150
IV. DNA Mediated Transformation 163
V. Summary and Prospects 173
References 176
v
PART II Methods
4. Tetrahymena as a Laboratory Organism: Useful Strains, Cell Culture, and
Cell Line Maintenance
Eduardo Orias, Eileen P. Hamilton, and Judith D. Chios
I. Introduction 190
II. Useful Tetrahymena Strains 190
III. Culture Media 194
IV. Culturing Tetrahymena Cells 199
V. Cell Line Storage 202
References 208
5. Long Term Storage
Peter]. Bruns, Helen R. Smith, and Donna Cassidy Hanley
I. Introduction 213
II. Supplies 214
III. Preparation of Cells before Freezing 215
IV. Freezing 216
V. Thawing and Recovery 216
VI. Conclusion 217
References 217
6. Genetic Crosses: Setting Up Crosses, Testing Progeny, and Isolating
Phenotypic Assonants
Eileen P. Hamilton and Eduardo Orias
I. Introduction 219
II. Methods for Mating Cells 220
III. How to Isolate and/or Select Progeny from a Cross 222
IV. Mating Type Tests and Sexual Immaturity Tests 225
V. Isolating Assonants 227
References 228
7. Methods for Genetic Analysis
Peter J. Bruns and Donna Cassidy Hanley
I. Introduction 229
II. Reagents 230
III. Equipment 231
IV. Strains 234
V. Strategies 235
References 240
8. Isolation of Micronuclear and Macronuclear DNA
Sally Lyman Allen
I. Introduction 242
II. Cell Culture 243
III. Percoll Method for Isolation of Nuclei 244
IV. Isolation of High Molecular Weight DNA from Nuclei 247
V. Solutions 249
References 252
9. Genetically Sorting a Collection of Tetrahymena Mutants
Eduardo Orias and Eileen P. Hamilton
I. Introduction 253
II. Mapping the Mutations to Chromosome Arms 255
III. Genetic Complementation Tests 255
References 263
10. Genetically Mapping New Mutants and Cloned Genes
Eileen P. Hamilton and Eduardo Orias
I. Introduction 265
II. Genetically Mapping a Mutation 266
III. Genetically Mapping a Cloned Gene or Sequence 278
References 279
11. Selection of Motility Mutants
David G. Pennock
I. Introduction 281
II. Materials 282
III. Special Procedures and Equipment 283
IV. Strains 283
V. Inducing and Screening for Temperature Sensitive Mutations Affecting
Cilia Regeneration 283
VI. Mutagenesis by Targeted Gene Knockout 286
References 289
12. Nuclear and Cytoskeletal Fluorescence Microscopy Techniques
K. R. Stuart and E. S. Cole
I. Introduction 291
II. DAPI: A Fluorescent Nuclear Stain 292
HI. Immunofluorescence Labeling for Conventional Fluorescence Microscopy 299
IV. Immunofluorescence Labeling for Confocal Fluorescence Microscopy 305
References 310
13. Nuclear and Cortical Histology for Brightfield Microscopy
E. S. Cole and K. R. Stuart
I. Introduction 313
II. Silver Staining 314
HI. Protargol Staining 317
IV. Giemsa Staining 319
References 321
14. Fixation of Tetrahymena Cells for Electron Microscopy
William Dentler
I. Introduction 323
II. CeU Culture 324
III. Fixation 324
IV. Critical Point Drying for SEM 327
V. Embedding for TEM 328
VI. Staining Sections (TEM) 330
References 331
15. Immunoelectron Microscopy of Tetrahymena
R. H. Gavin, John G. Hoey, and Jorge A. Garces
I. Introduction 334
II. General Considerations 334
III. Postembedding Labeling 336
IV. Double Labeling with the Postembedding Technique 338
V. Pre embedding Labeling 340
VI. Controls for Immunogold Labeling 340
VII. Quantitative Analysis of Colloidal Gold Labeling 341
VIII. Troubleshooting: Problems and Recommended Remedies 342
References 342
PART III CeU Biology
16. Regulated Protein Secretion in Tetrahymena thermophila
Aaron P. Turkewitz, N. Doane Chilcoat, Alex Haddad, and John W. Verbsky
I. Introduction 347
II. Isolation of the Released Contents of Dense Core Secretory Vesicles
(Mucocysts) 350
III. Stimulation of Cells to Test Exocytic Capability or to Study Postexocytic
Events 354
IV. Isolation of Intact Secretory Vesicles 357
References 360
17. Electrophysiology of Tetrahymena
Todd M. Hennessey and Heather G. Kuruvilla
I. Introduction 363
II. Materials and Methods 367
III. Commentary and Summary 374
References 376
18. Isolation and Characterization of in Vivo Modified Histones and an
Activity Gel Assay for Identification of Histone Acetyltransferases
Emily A. Wiley, Craig A. Mizzen, and C. David AHis
I. Introduction 379
II. Methods 382
References 392
19. Tetrahymena Telomerase Activity, Purification, and Reconstitution
Chantal Autexier
I. Introduction 396
II. Materials and Methods 400
III. Commentary and Summary 409
References 412
20. Studying the Telomerase RNA in Tetrahymena
E. Blackburn, D. Gilley, T. Ware, A. Bhattacharyya, K. Kirk, and H. Wang
I. Introduction 417
II. General Strategy and Overview 419
III. Basic Methods 420
References 431
21. Isolation and Characterization of 22S Outer Arm Dynein from
Tetrahymena Cilia
Trade M. Gibson and David J. Asai
I. Introduction 433
II. Methods 435
III. Summary 438
References 439
22. Preparation of Cytoskeletal Fractions from Tetrahymena thermophila
Norman E. Williams
I. Introduction 441
II. Methods 442
References 446
23. Immunoprecipitation Procedures
Norman E. Williams
I. Introduction 449
II. Method 450
References 453
24. Tetmhymena Calcium Binding Proteins, TCBP 25 and TCBP 23
Osamu Numata, Kazuko Hanyu, Tetsuya Takeda, and Yoshio Watanabe
I. Introduction 455
II. Methods 456
References 465
PART IV Manipulating Genes
25. Microinjection of Tetmhytnena thermophila
Douglas L. Chalker, John G. Ward, Caterina Randolph, and Meng Chao Yao
I. Introduction 469
II. The Technique of Microinjection 472
III. Concluding Remarks 482
References 483
26. Transient and Stable DNA Transformation of Tetrahymena thermophila
by Electroporation
Jacek Gaertig and Geoffrey Kapler
I. Introduction 486
II. Considerations 487
III. Preparing Plasmid DNA for Electrotransformation 488
IV. General Protocol for Conjugant Electrotransformation 489
V. Transformation with Integrating Vectors: Gene Replacements and Gene
Knockouts 492
VI. Transformation with Autonomously Replicating Vectors 495
References 498
27. Biolistic Transformation of Macro and Micronuclei
Peter J. Bruns and Donna Cassidy Hanley
I. Introduction 501
II. Cells 502
III. Transformation 504
References 511
28. Knockout Heterokaryons Enable Facile Mutagenic Analysis of Essential
Genes in Tetrahymena
Bing Hai, Jacek Gaertig, and Martin A. Gorovsky
I. Introduction 514
II. Strategy 515
HI. Methods for Creating Knockout Heterokaryons 524
References 530
29. Creation and Use of Antisense Ribosomes in Tetrahymena thermophila
Qichang Fan, Rosemary Sweeney, and Meng Chao Yao
I. Introduction 533
II. Using Antisense Ribosomes in Tetrahymena thermophila 534
III. Considerations Particular to Antisense Ribosomes 538
IV. Evaluation and Applications of the Antisense Ribosome System 544
References 545
30. Protein Tagging in Tetrahymena
Lanlan Yu and Martin A. Gorovsky
I. Introduction 549
II. Strategies for Gene Tagging in Tetrahymena 551
III. Methods 557
References 557
Appendix I. Genetic Nomenclature Rules for Tetrahymena thermophila 561
Appendix II. Codon Usage in Tetrahymena thermophila 565
Index 569
Volumes in Series 575
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| series | Methods in cell biology |
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| spelling | Tetrahymena thermophila ed. by David J. Asai ... San Diego [u.a.] Acad. Press 2000 XVIII, 580 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in cell biology 62 Protozoa gtt Tetrahymena Cell Culture Techniques Tetrahymena thermophila Tetrahymena thermophila (DE-588)4274357-6 gnd rswk-swf Cytologie (DE-588)4070177-3 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Tetrahymena thermophila (DE-588)4274357-6 s Cytologie (DE-588)4070177-3 s DE-604 Asai, David J. Sonstige oth Methods in cell biology 62 (DE-604)BV002534878 62 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008728913&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Tetrahymena thermophila Methods in cell biology Protozoa gtt Tetrahymena Cell Culture Techniques Tetrahymena thermophila Tetrahymena thermophila (DE-588)4274357-6 gnd Cytologie (DE-588)4070177-3 gnd |
| subject_GND | (DE-588)4274357-6 (DE-588)4070177-3 (DE-588)4143413-4 |
| title | Tetrahymena thermophila |
| title_auth | Tetrahymena thermophila |
| title_exact_search | Tetrahymena thermophila |
| title_full | Tetrahymena thermophila ed. by David J. Asai ... |
| title_fullStr | Tetrahymena thermophila ed. by David J. Asai ... |
| title_full_unstemmed | Tetrahymena thermophila ed. by David J. Asai ... |
| title_short | Tetrahymena thermophila |
| title_sort | tetrahymena thermophila |
| topic | Protozoa gtt Tetrahymena Cell Culture Techniques Tetrahymena thermophila Tetrahymena thermophila (DE-588)4274357-6 gnd Cytologie (DE-588)4070177-3 gnd |
| topic_facet | Protozoa Tetrahymena Cell Culture Techniques Tetrahymena thermophila Cytologie Aufsatzsammlung |
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