mRNA formation and function:
Gespeichert in:
Format: | Buch |
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Sprache: | English |
Veröffentlicht: |
San Diego [u.a.]
Acad. Press
1997
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Schlagworte: | |
Online-Zugang: | Inhaltsverzeichnis |
Beschreibung: | XVII, 395 S. Ill., graph. Darst. |
ISBN: | 0125875452 |
Internformat
MARC
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245 | 1 | 0 | |a mRNA formation and function |c ed. by Joel D. Richter |
264 | 1 | |a San Diego [u.a.] |b Acad. Press |c 1997 | |
300 | |a XVII, 395 S. |b Ill., graph. Darst. | ||
336 | |b txt |2 rdacontent | ||
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Datensatz im Suchindex
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adam_text | CONTENTS
Contributors xiii
Preface xvii
CHAPTER 1
The Tat TAR RNP, a Master Switch That Regulates
HIV 1 Gene Expression
Carlos Sunk, Paul R. Bohjanen, Yi Liu, and Mariano A. Garcia Blanco
I. Introduction 1
II. Methods 4
References 21
CHAPTER 2
HeLa Nuclear Extract: A Modified Protocol
Cameron R. Miller, Sharon F. Jamison, and Mariano A. Garcia Blanco
I. Growth of the HeLa Cells 25
II. Extract Preparation 26
Reference 30
CHAPTER 3
Functional Analysis of Splicing Factors
and Regulators
Juan Valcarcel, Conception Martinez, and Michael R. Green
I. Introduction 31
II. Functional Analysis of General Splicing Factors 32
III. Analysis of Regulatory Mechanisms in
Pre mRNA Splicing 43
References 49
V
vi Contents
CHAPTER 4
Identification of Pre mRNA Splicing Factors and
Analysis of RNA—Protein Interaction
James G. Pattern, Billy T. Dye, Daron C. Barnard, and James G. McAfee
I. Introduction 55
II. Pre mRNA Splicing and the Role of the
Polypyrimidine Tract 56
III. Analysis of RNA Binding Proteins by
UV Cross Linking 58
¦ IV. Analysis of Protein RNA Interaction by
Fluorescence Spectroscopy 62
V. Analysis of Pre mRNA Splicing Complexes and
Identification of Associated Factors by
UV Cross Linking 68
VI. Identification of RNA Binding Proteins by a cDNA
Expression Library Screen 72
References 75
CHAPTER 5
In Vitro Analysis of Mammalian Cell mRNA
3 Processing
Gregory M. Gilmartin
I. Introduction 79
II. General Considerations for the Analysis of mRNA 3
mRNA Processing in Vitro 80
III. Pre mRNA Cleavage and Polyadenylation 82
IV. 3 Processing Complex Formation 85
V. 3 Processing Complex Stability 88
VI. Probing 3 Processing Factor Pre mRNA Interactions
by UV Cross Linking 90
VII. Application of in Vitro Selection to the Analysis of
Sequences That Direct 3 Processing 93
VIII. Conclusions 97
References 97
Contents vii
CHAPTER 6
Rapid Identification and Cloning of
Sequence Specific RNA Binding Proteins
Jeffrey Wilmz
I. Introduction 99
II. Preparation of Extracts 100
III. Preparation of RNAs 101
IV. Identification of Protein RNA Complexes 102
V. Determination of RNA Sequence Requirements for
Protein Binding 103
VI. Cloning cDNAs for RNA Binding Proteins by
Northwestern Screening 105
References 108
CHAPTER. 7
Analysis of Polyadenylation Phenotypes in
Saccharomyces cerevisiae
J. Scott Butler, Michael W. Briggs, and Aaron Proweller
I. Introduction 111
II. Poly(A) Tail Analysis 112
III. Polyadenylation Site Mapping by RT PCR 117
IV. Preparation of Yeast mRNA 3 End
Processing Extracts 121
References 125
CHAPTER 8
Poly(A) Polymerase/Cap Specific
2 O Methyltransferase from Vaccinia Virus:
Expression, Purification, Uses, and Protein—Ligand
Interaction Assays
Paul D. Gershon
I. Background 127
II. Expression and Purification of the Poly(A) Polymerase/
2 O Methyltransferase 132
III. Uses of the Poly(A) Polymerase/2 O Methyltransferase 135
IV. Assays Developed to Examine Protein Ligand
Interactions 139
References 145
Vlii Contents
CHAPTER 9
A Genetic Approach to Mapping Coding Region
Determinants ofmRNA Instability in Yeast
Aidan N. Hennigan and Allan Jacobson
I. Introduction 149 j
II. Experimental Methodology 150 i
HI. Experimental Results 153 ¦
IV. Discussion and Conclusions 157 !
References 160
CHAPTER 10
Identification of the Protein That Interacts with the
3 End of Histone mRNA
William F. Marzluff, Michael L. Whitfield, Zbigniew Dotninski, and Zeng Feng Wang
I. Introduction 163
II. Detecting Specific Protein RNA Interactions 165
III. In Vitro Processing of Histone Pre mRNA 174
IV. Isolation of Polyribosomes 181
V. Isolation and Characterization of RNA Binding Proteins
Using the Yeast Three Hybrid System 183
Appendix: Cell Culture 188
References 192
CHAPTER 11
Analysis of mRNP Complexes Assembled in Vitro
Martin Holcik and Stephen A. Uebhaber
I. Complex Formation 196
II. Detection of RNP Complexes 200
HI. Analysis of RNP Complexes Identified by
Electrophoretic Mobility Shift Analysis 202
References 208
Contents ix
CHAPTER 12
Analysis of RNA Binding Proteins Using
in Vitro Genetics
he A. Laird Offringa
I. Introduction 212
II. Successfully Making Display Phage 215
HI. Binding of Displayed RNA Binding Proteins to RNA 221
IV. Application of in Vitro Genetics to the Study of RNA
Binding Proteins 224
V. Protocols 227
References 234
CHAPTER 13
Interactions of Proteins with Specific Sequences
in RNA
Lucy G. Andrews and Jack D. Keene
I. Introduction 237
II. Selection of Specific Sequences from Random RNA
Libraries by RNA Binding Proteins as a First Step in
Identification of in Vivo Targets 240
III. Binding of Protein to Sequences in Cellular RNAs 247
IV. Determination of Minimal Binding Sequences within
RNA Ligands 250
V. Selection of RNAs from 3 UTR Libraries 257
VI. Summary 259
References 259
CHAPTER 14
A Rapid Genetic Method for the Study of RNA
Binding Proteins
Chaitanya Jain and Joel G. Belasco
I. Introduction 263
II. Background 264
III. A Quantitative Model for Translation Repression by
Heterologous RNA BPs 265
IV. Methods 267
V. Strategies and Applications 274
X Contents
VI. Troubleshooting 280
VII. Application of the Genetic Methods to Rev 281
VIII. Conclusions 282
References 283
CHAPTER 15
Footprinting RNA Protein Complexes with
Hydroxyl Radicals
Patricia Ansel McKinney and Lee Gehrke
I. Introduction 286
II. Preparing an RNA Template for Hydroxyl
Radical Footprinting 288
III. Binding and Cleavage Reactions 292
IV. Footprinting Data 293
V. Concluding Remarks 299
References 300
CHAPTER 16
Analysis of Ribosome Loading onto mRNA Species:
Implications for Translational Control
Hangjun Ruan, Cheryl Y. Brown, and David R. Morris
I. Polysome Profiles and Translational Control 305
II. Extraction and Display of Polysomes 307
III. RNA Analysis by Northern Blots 312
IV. Analysis by RNase Protection 315
References 320
CHAPTER 17
Analysis of Picornavirus Internal Ribosome Entry
Site Function in Vivo
Graham J. Belsham
I. Introduction 323
II. Cap Dependent and Cap Independent Translation 324
III. Characteristics of Picornavirus IRES Elements 325
Contents xi
IV. Characteristics of the Vaccinia/T7 RNA Polymerase
Expression System 327
V. Analysis of Picornavirus IRES Function in Vivo 328
VI. Distinguishing between Cap Dependent and IRES
Directed Translation 334
VII. Complementation of Defective IRES Elements by Co
Expression of the wt IRES in Trans 335
References 337
CHAPTER 18
RNA Traffic and Localization Reported by
Fluorescent Molecular Cytochemistry in
Living Cells
Marty R. Jacobson and Tlioru Pederson
I. Introduction 341
II. Fluorescent Labeling of RNA 342
III. Cell Culture, Microinjection, and Digital
Imaging Processing 344
IV. Examples 350
V. What Is the Biological Authenticity of
Fluorescent RNA? 351
VI. Technical Problems and Their Resolution 353
VII. Integrating Fluorescent RNA Cytochemistry with
Other Methods 354
VIII. What Does Fluorescent RNA Cytochemistry
Actually Measure? 355
IX. New Frontier Methods 356
X. Closing Remarks 356
References 357
CHAPTER. 19
Detection of mRNA in Situ: Techniques
for Studying Gene Expression in
Drosophila melanogaster Tissues
Armen S. Manoukian
I. Introduction 361
II. Whole Mount in Situ Hybridization to mRNA with
DIG Labeled Probes 362
xii Contents
III. Uses of the Whole Mount Procedure 365
IV. Detection of Multiple Gene Products 367
V. Subcellular Localization of unngless niRNA 369
References 369
CHAPTER 20
Differential Display Protocol That Preferentially
Identifies mRNAs of Moderate to Low Abundance
in a Microscopic System
Ognian C. Ikonomov and Michele H. Jacob
I. Introduction 371
II. Experimental Procedures 373
III. Confirmation of Differential Expression 379
IV. Concluding Remarks 381
References 382
Index 385
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isbn | 0125875452 |
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spelling | mRNA formation and function ed. by Joel D. Richter San Diego [u.a.] Acad. Press 1997 XVII, 395 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier ARN messager ARN messagers ram Messenger RNA Research Methodology RNA, Messenger Research methods Messenger-RNS (DE-588)4169499-5 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Messenger-RNS (DE-588)4169499-5 s Methode (DE-588)4038971-6 s DE-604 Richter, Joel D. Sonstige oth HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007812462&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
spellingShingle | mRNA formation and function ARN messager ARN messagers ram Messenger RNA Research Methodology RNA, Messenger Research methods Messenger-RNS (DE-588)4169499-5 gnd Methode (DE-588)4038971-6 gnd |
subject_GND | (DE-588)4169499-5 (DE-588)4038971-6 (DE-588)4143413-4 |
title | mRNA formation and function |
title_auth | mRNA formation and function |
title_exact_search | mRNA formation and function |
title_full | mRNA formation and function ed. by Joel D. Richter |
title_fullStr | mRNA formation and function ed. by Joel D. Richter |
title_full_unstemmed | mRNA formation and function ed. by Joel D. Richter |
title_short | mRNA formation and function |
title_sort | mrna formation and function |
topic | ARN messager ARN messagers ram Messenger RNA Research Methodology RNA, Messenger Research methods Messenger-RNS (DE-588)4169499-5 gnd Methode (DE-588)4038971-6 gnd |
topic_facet | ARN messager ARN messagers Messenger RNA Research Methodology RNA, Messenger Research methods Messenger-RNS Methode Aufsatzsammlung |
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