Small GTPases of the Ras superfamily are molecular switches which cycle between an active guanosine triphosphate (GTP)-bound and an inactive guanosine diphosphate (GDP)-bound state. They integrate signals from the cell surface to the nucleus, regulating important cellular activities. For example, Ras itself is activated when extracellular growth factors such as platelet derived growth factor (PDGF) or epidermal growth facor (EGF) bind to their receptors at the cell surface. This activation of Ras ultimately leads to changes in the transcriptional activity of the cell, e.g., via the canonical mitogen activated protein kinase (MAPK) cascade. Constitutively activated, mutant forms of Ras such as RasV12 are found frequently in human tumors, and it is widely assumed that this oncogene acts via transcriptional activation of growth and proliferation pathways.
- Polymerase Chain Reaction
- Suppression Subtractive Hybridization
- Polymerase Chain Reaction Buffer
- Polymerase Chain Reaction Cycle
- Polymerase Chain Reaction Tube
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, access via your institution.
Tax calculation will be finalised at checkout
Purchases are for personal use onlyLearn about institutional subscriptions
Van Aelst, L. and D’Souza-Schorey, C. (1997) Rho GTPases and signaling networks. Genes Dev. 11, 2295–2322.
Velculescu, V. E., Zhang, L., Vogelstein, B., and Kinzler, K. W. (1995) Serial analysis of gene expression. Science 270, 484–487.
Liang, P. and Pardee, A. B. (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257, 967–971.
Liang, P. and Pardee, A. B., eds. (1997) Differential Display Methods and Protocols. Methods Mol. Biol., Vol. 85, Humana Press, Totowa, NJ, pp. 1–320.
Hubank, M. and Schatz, D. G. (1994) Identifying differences in mRNA expression by representational difference analysis of cDNA. Nucleic Acids Res. 22, 5640–5648.
Hubank, M. and Schatz, D. G. (1999) cDNA representational difference analysis: a sensitive and flexible method for identification of differentially expressed genes. Methods Enzymol. 303, 325–349.
Diatchenko, L., Lau, Y. F., Campbell, A. P., Chenchik, A., Moqadam, F., Huang, B., et al. (1996) Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc. Natl. Acad. Sci. USA 93, 6025–6030.
Zuber, J., Tchernitsa, O. I., Hinzmann, B., Schmitz, A. C., Grips, M., Hellriegel, M., et al. (2000) A genome-wide survey of RAS transformation targets. Nat. Genet. 24, 144–152.
Harris, A. J., Shaddock, J. G., Manjanatha, M. G., Lisenbey, J. A., and Casciano, D. A. (1998) Identification of differentially expressed genes in aflatoxin B1-treated cultured primary rat hepatocytes and Fischer 344 rats. Carcinogenesis 19, 1451–1458.
Phimister, B., ed. (1999) The Chipping Forecast. Nat. Genet. 21(Suppl.), 1–60.
Lisitsyn, N., Lisitsyn, N., and Wigler, M. (1993) Cloning the differences between two complex genomes. Science 25, 946–951.
Lisitsyn, N. and Wigler, M. (1995) Representational difference analysis in detection of genetic lesions in cancer. Methods Enzymol. 254, 291–304.
Welford, S. M., Gregg, J., Chen, E., Garrison, D., Sorensen, P. H., Denny, C. T., and Nelson, S. E (1998) Detection of differentially expressed genes in primary tumor tissues using representational differences analysis coupled to microarray hybridization. Nucleic Acids Res. 26, 3059–3065.
Clackson, T. (1997) Controlling mammalian gene expression with small molecules. Curr. Opin. Chem. Biol. 1, 210–218.
No, D., Yao, T. P., and Evans, R. M. (1996) Ecdysone-inducible gene expression in mammalian cells and transgenic mice. Proc. Natl. Acad. Sci. USA 93, 3346–3351.
Gossen, M. and Bujard, H. (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA 89, 5547–5551.
Jou, T. S. and Nelson, W. J. (1998) Effects of regulated expression of mutant RhoA and Rac1 small GTPases on the development of epithelial (MDCK) cell polarity. J. Cell Biol. 142, 85–100.
Lee, J. C., Laydon, J. T., McDonnell, P. C., Gallagher, T. F., Kumar, S., Green, D., et al. (1994) A protein kinase involved in the regulation of inflammatory cytokine biosynthesis. Nature 372, 739–746.
Lockhardt, D. J. and Winzeler, E. A. (2000) Genomics, gene expression, and DNA arrays. Nature 405, 827–836.
Editors and Affiliations
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Schmitz, A.A.P., Lucito, R., Aelst, L.V. (2002). Using cDNA-Representational Difference Analysis (cDNA-RDA) in Combination with Microarrays to Identify Rac Regulated Genes. In: Manser, E., Leung, T. (eds) GTPase Protocols. Methods in Molecular Biology™, vol 189. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-281-3:025
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-934-6
Online ISBN: 978-1-59259-281-4
eBook Packages: Springer Protocols