Abstract
In recent years, interest in RNA-binding proteins (RNA-BPs) and RNA-protein interactions has grown considerably. An important reason for this interest is the increased recognition that RNA-BPs are involved in a wide variety of critical viral and cellular processes including transcription, translation, RNA processing, RNA transport, gene regulation, and splicing. A second reason lies in the technical advances and the generation of new methods that have simplified the analysis of RNA-protein interactions. Nevertheless, a thorough analysis of an RNA-BP is not a trivial task, and, at present, there are only a few RNA-BPs that have been characterized in detail.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Jain, C. and Belasco, J. G. (1996) A structural model for the HIV-1 Rev-RRE complex deduced from the altered specificity of Rev variants isolated by a rapid genetic screening strategy. Cell 87, 115–125.
Bouvet, P., Jain, C., Belasco, J. G., Almaric, F., and Erard, M. (1997) RNA recognition by the joint action of two nucleolin RNA-binding domains: genetic analysis and structural modeling. EMBO J. 16, 5235–5246.
Dreyfus, M. (1988) What constitutes the signal for the initiation of protein synthesis on E. coli mRNAs? J. Mol. Biol. 204, 79–94.
Gold, L. (1988) Posttranscriptional regulatory mechanisms in Escherichia coli. Annu. Rev. Biochem. 57, 199–233.
de Smit, M. H. and van Duin, J. (1990) Secondary structure of the ribosome binding site determines translational efficiency: a quantitative analysis. Proc. Natl. Acad. Sci. USA 87, 7668–7672.
Rose, R. E. (1988) The nucleotide sequence of pACYC184. Nucleic Acids. Res. 16, 355.
Miller, J. H. (1972) in Experiments in Molecular Genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Peabody, D. (1993) The RNA binding site of bacteriophage MS2 coat protein. EMBO J. 12, 595–600.
Kunkel, T. A., Bebenek, K., and McClary, J. (1991) Efficient site-directed mutagenesis using uracil-containing DNA. Methods Enzymol. 204, 125–139.
Olsen, D. B., Sayers, J. R., and Eckstein, F. (1993) Site-directed mutagenesis of single-stranded and double-stranded DNA by phosphorothionate approach. Methods Enzymol. 217, 189–217.
Cadwell, R. C. and Joyce, G. F. (1992) Randomization of genes by PCR mutagenesis. PCR Methods Appl. 2, 28–33.
Jain, C. and Belasco, J. G. (1997) A genetic method for the study of RNA-binding proteins, in mRNA Formation and Function (Richter, J., ed.), Academic Press, New York, NY, pp. 263–284.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
Jain, C. (1999). An Escherichia coli-Based Genetic Strategy for Characterizing RNA Binding Proteins. In: Haynes, S.R. (eds) RNA-Protein Interaction Protocols. Methods in Molecular Biology™, vol 118. Humana Press. https://doi.org/10.1385/1-59259-676-2:161
Download citation
DOI: https://doi.org/10.1385/1-59259-676-2:161
Publisher Name: Humana Press
Print ISBN: 978-0-89603-568-3
Online ISBN: 978-1-59259-676-8
eBook Packages: Springer Protocols