UID:
almahu_9947366700102882
Format:
1 online resource (621 p.)
ISBN:
1-282-28732-X
,
9786612287329
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0-08-092206-6
Series Statement:
Methods in enzymology ; v. 447
Content:
Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements, responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation.〈br
Note:
Description based upon print version of record.
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Front Cover; Methods in Enzymology; Copyright Page; Contents; Contributors; Preface; Volumes in Series; Section I: Bacteria; Chapter 1: Analysis of RNA Decay, Processing, and Polyadenylation in Escherichia Coli and Other Prokaryotes; 1. Introduction; 2. General Considerations When Working with RNA; 3. Isolation of Total RNA from Exponentially Growing Cells; 4. Isolation of Total RNA from Stationary Phase Cells; 5. Removal of DNA Contamination; 6. RNA Yield and Purity Assessment; 10. Analysis of RNA Polyadenylation in E. Coli; 12. Conclusion
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Chapter 2: Analyzing the Decay of Stable RNAs in E. Coli1. Introduction; 2. Preparation of Stable RNA Substrates for in Vitro Degradation Assays; 3. Detection of Degradation Products in Vitro; 6. Detection of Poly(A) Tails on Products Generated by Processing or Degradation of Stable RNA; Acknowledgments; References; Chapter 3: Genomic Analysis of mRNA Decay in E. Coli with DNA Microarrays; 1. Introduction; 4. Data Acquisition and Analysis; Chapter 4: Co-immunopurification of Multiprotein Complexes Containing RNA-Degrading Enzymes; 1. Introduction; 4. Protocols; 5. Perspective; References
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Chapter 5: PABLO Analysis of RNA: 5prime-Phosphorylation State and 5prime-End Mapping1. Introduction; 4. Efficiency of Ligation; 6. Ascertaining Whether the Decay of a Transcript Begins with Pyrophosphate Removal; 7. PABLO Protocols; 8. Alternative Method for Examining the Phosphorylation State of RNA; References; Chapter 6: A Proteomic Approach to the Analysis of RNA Degradosome Composition in Escherichia coli; 1. Introduction; 2. Experimental Procedures; Acknowledgments; References
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Chapter 7: New Approaches to Understanding Double-Stranded RNA Processing by Ribonuclease III: Purification and Assays of Homodimeric and Heterodimeric Forms of RNase III from Bacterial Extremophiles and Mesophiles1. Introduction; 2. Heterologous Expression, Affinity Purification, and Assays of Aquifex aeolicus and Thermotoga maritima RNases III; 3. Production and Purification of Escherichia Coli RNase III Heterodimers; References; Chapter 8: Characterizing Ribonucleases In Vitro: Examples of Synergies between Biochemical and Structural Analysis; 1. Introduction
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3. Construction of RNase II Derivatives5. Analysis of Ribonucleolytic Activity; 6. Studying the RNA-Binding Abilities; 7. Structural Studies of RNase II; References; Chapter 9: The Role of RNA Chaperone Hfq in Poly(A) Metabolism: Methods to Determine Positions, Abundance, and Lengths of Short Oligo(A) Tails; 1. Introduction; 3. RT-PCR-Based Analysis of 3prime-RNA Ends; 4. Effect of Hfq on Polyadenylation of the rpsO Transcript; 6. Hfq Stimulates Poly(A) Synthesis; Acknowledgments; References; Chapter 10: Assaying DEAD-box RNA Helicases and Their Role in mRNA Degradation in Escherichia coli
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1. Introduction
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English
Additional Edition:
ISBN 0-12-374377-X
Language:
English
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