UID:
almafu_9958128399702883
Umfang:
1 online resource (420 p.)
ISBN:
9786611766221
,
9781281766229
,
1281766224
,
9780080889665
,
0080889662
Serie:
Methods in enzymology, v. 429
Inhalt:
For over fifty years the Methods in Enzymology series has been the critically aclaimed laboratory standard and one of the most respected publications in the field of biochemistry. The highly relevant material makes it an essential publication for researchers in all fields of life and related sciences. This volume, the first of three on the topic of Translation Initiation includes articles written by leaders in the field.
Anmerkung:
Description based upon print version of record.
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Cover; Copyright Page; TOCContents; Contributors; Preface; Volumes in Series; CHChapter 1: Use of Reticulocyte Lysates for Mechanistic Studies of Eukaryotic Translation Initiation; 1. Introduction; 2. Materials; 2.1. Nuclease-treated reticulocyte lysates; 2.2. tRNA; 2.3. mRNA; 3. Methods; 3.1. Preparation of tRNA; 3.2. Preparation of mRNA; 4. Translation of an mRNA to Yield a Radioactive Product; 5. Quantitation of Reaction Products; 6. Optimization of Translations; 7. Reporter Proteins for Translation; 8. Experimental Use of Nuclease-Treated Lysates; 8.1. Initiation mechanisms
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8.2. Competition between mRNAs8.3. Synthesis of proteins of high specific radioactivity; 8.4. Influence of variations of factor activity; 8.5. Sucrose gradients; 8.6. Toe printing; References; CHChapter 2: Studying Translational Control in Drosophila Cell-Free Systems; 1. Introduction; 2. Preparation of Ovary Extracts; 3. Preparation of Embryo Extracts; 4. The Translation Assay; Acknowledgments; References; CHChapter 3: Use of In Vitro Translation Extract Depleted in Specific Initiation Factors for the Investigation of Translational Regulation; 1. Introduction
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2. Factors Involved in Translation Initiation3. Experimental Methods to Generate and Use Fractionated Translation Extracts; 3.1. In vitro RNA synthesis; 3.2. Preparation of fractionated lysates; 3.3. SDS-PAGE analysis; 3.4. Western analysis; 3.5. In vitro translation in fractionated lysates; References; CHChapter 4: A Highly Efficient and Robust In Vitro Translation System for Expression of Picornavirus and Hepatitis C Virus RNA Genomes; 1. Introduction; 2. Cell-Free Model for EMCV Replication; 2.1. Picornavirus replication: An overview; 2.2. Experimental results
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3. Materials for Cell-Free Synthesis of EMCV3.1. Animals, cells, virus, and viral RNA; 3.2. Media and solutions; 3.3. Other materials; 4. Methods for Cell-Free Synthesis of EMCV; 4.1. Propagation and storage of Krebs-2 cells; 4.2. Preparation of Krebs-2 S10 extract; 4.3. Nuclease treatment of the extract; 4.4. Translation protocol; 4.5. Analyzing translation; 4.6. EMCV RNA replication protocol; 4.7. Analyzing RNA replication; 4.8. Virus synthesis protocol; 4.9. Plaque assay for infectivity; 5. In Vitro Translation of HCV RNA; 5.1. HCV replication: An overview
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5.2. General characteristics of HCV RNA-directed translation and polyprotein processing in vitro6. Materials for In Vitro Translation of HCV RNA; 7. Methods and Applications of In Vitro Translation of HCV RNA; 7.1. HCV RNA in vitro translation protocol; 7.2. Characterizing NS3 protease inhibitors; 7.3. Probing glycosylation of HCV envelope proteins; 7.4. Protease protection assay for translocation of HCV envelope proteins; 8. Perspectives and Future Applications; Acknowledgments; References; CHChapter 5: A Practical Approach to Isolate 48S Complexes: Affinity Purification and Analyses
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1. Introduction
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English
Weitere Ausg.:
ISBN 9780123741912
Weitere Ausg.:
ISBN 0123741912
Sprache:
Englisch
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