Abstract
The detection of gene expression can be accomplished in several ways. “Classical” methods (still important, of course) relied on the observation of biochemical or phenotypic changes in cells or organisms in order to determine the expression of a specific gene. Later, advances in macromolecular separation technology made feasible the identification and isolation of a particular gene product or protein molecule. With the advent of recombinant DNA techniques, it is now possible to detect and analyze the transcriptional product of any gene. There are several methods now in widespread use for studying specific RNA molecules. These methods include in situ hybridization,1 Northern gels,2 dot-or slot-blots,2,3 S-1 nuclease assays,4 and RNase protection studies.5 In this chapter, we describe a new and powerful method to detect gene expression at the RNA level. Protocols will be outlined and discussed in detail and some applications will be briefly described. No attempt has been made to exhaustively review the literature in this fast moving area, so we wish to apologize beforehand to the numerous authors whose work may have not been cited in this chapter.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Angerer, R.C., Cox, K.H., and Angerer, L.M. (1985) Genetic Engineering 7:43–65.
Thomas, P.S. (1980) Proc. Natl. Acad. Sci. USA 77:5201–5205.
White, B.A., and Bancroft, F.C. (1982) J. Biol. Chem. 257:8569–8572.
Berk, A.J., and Sharp, P.A. (1977) Cell 12:721–732.
Melton, D.A., Krieg, P.A., Rabagliati, M.R., Maniatis, T., Zinn, K., and Green, M.R. (1984) Nucl. Acids Res. 7:1175–1193.
Davis, L.G., Dibner, M.D., and Battey, J.F. (1986) Basic Methods in Molecular Biology. Elsevier, N.Y.
Ausubel, F.M., Brent, R., Kingston, R.F., Moore, D.D., Seidman, J.G., Smith, J.A., and Struhl, K. (1987) Current Protocols in Molecular Biology. John Wiley & Sons, N.Y.
Berger, S.L., and Kimmel, A.R. (1987) Guide to Molecular Cloning Techniques. Methods in Ezymology, Vol. 152, Academic Press, Inc., Orlando.
Kawasaki, E.S., Clark, S.S., Coyne, M.Y., Smith, S.D., Champlin, R., Witte, O.N., and McCormick, F.P. (1988) Proc. Natl. Acad. Sci. USA 85:5698–5702.
Noonan, K.E., and Roninson, I.B. (1988) Nucl. Acids Res. 16:10366.
Veres, G., Gibbs, RA., Scherer, S.E., and Caskey, C.T. (1987) Science 237: 415–417.
Hawkins, J.D. (1988) Nucl. Acids Res. 16:9893–9908.
Rappolee, D.A., Mark, D., Banda, MJ., and Werb, Z. (1988) Science 241:708–712.
Chelly, J., Kaplan, J.-C., Maire, P., Gautron, S., and Kahn, A. (1988) Nature 333:858–860.
Harbarth, P., and Vosberg, H.-P. (1988) DNA 7:297–306.
Lee, M-S., Chang, K.S., Freireich, E.J., Kantarjian, H.M., Talpaz, M., Trujillo, J.M., and Stass, SA. (1988) Blood 72:893–897.
Price, C.M., Rassool, S., Shivji, M.K.K., Gow, J., Tew, CJ., Haworth, C., Goldman, J.M., and Wiedemann, L.M. (1988) Blood 72:1829–1832.
Dobrovic, A., Trainor, J., and Morley, A.A. (1988) Blood 72:2063–2065.
Hermans, A., Selleri, L., Gow, J., and Grosveld, G.C. (1988) Blood 72:2066–2069.
Kashani-Sabet, M., Rossi, J.J., Lu, U., Ma, J.X., Chen, J., Miyachi, H., and Scanlon, K.J. (1988) Cancer Res. 48:5775–5778.
Murakawa, GJ., Zaia, J.A., Spallone, P.A., Stephens, D.A., Kaplan, B.E., Wallace, R.B., and Rossi, J.J. (1988) DNA 7:287–295.
Byrne, B.C., Li, J.J., Sninsky, J., and Poiesz, B.J. (1988) Nucl. Acids Res. 16:4165.
Hart, C., Schochetman, G., Spira, T., Lifson, A., Moore, J., Galphiry J., Sninsky, J., and Ou, C.-Y. (1988) The Lancet II:596–599.
Gama, R.E., Hughes, P.J., Bruce, C.B., and Stanway, G. (1988) Nucl. Acids Res. 16:9346.
Lacal, J.C. and Tronick, S.R. (1988) in The Oncogene Handbook. Reddy, E.P., Skalka, A.M., and Curran, T., eds. Elsevier Science Publishers B.V., Amsterdam, pp. 257–304.
Powell, L.M., Wallis, S.C., Pease, RJ., Edwards, Y.H., Knott, TJ., and Scott, J. (1987) Cell 50:831-840.
Todd, JA., Bell, J.I., and McDevitt, H.O. (1987) Nature 329:599–604.
Sinha, A.A., Brautbar, C., Szafer, F., Friedmann, A., Tzfoni, E., Todd, J.A., Steinman, L., and McDevitt, H.O. (1988) Science 239:1026–1029.
Simpson, D., Crosby, R.M., and Skopek, T.R. (1988) Biochem. Biophys. Res. Comm. 151:487–492.
Acha-Orbea, H., Mitchell, D.J., Timmermann, L., Wraith, D.C., Tausch, G.S., Waldor, M.K., Zamvil, S.S., McDevitt, H.O., and Steinman, L. (1988) Cell 54:263–273.
Weiss, M.J., Cole, D.E., Ray, K., Whyte, M.P., Lafferty, M.A., Mulivor, R.A., and Harris, H. (1988) Proc. Natl. Acad. Sci. USA 85:7666–7669.
Hsu, T.-Y., Moroy, T., Etiemble, J., Louise, A., Trepo, C., Tiollais, P., and Buendia, M:-A. (1988) Cell 55:627–635.
Conboy, J.G., Chan, J., Mohandas, N., and Kan, Y.W. (1988) Proc. Natl. Acad. Sci. USA 85:9062–9065.
Lee, C.C., Wu, X., Gibbs, R.A., Cook, R.G., Muzny, D.M., and Caskey, C.T. (1988) Science 239:1288–1291.
Knoth, K., Roberds, S., Poteet, C., and Tamkun, M. (1988) Nucl. Acids. Res. 16:10932.
Frohman, M.A., Dush, M.K., and Martin, G.R. (1988) Proc. Natl. Acad. Sci. USA 85:8998–9002.
Sarkar, G., and Sommer, S.S. (1988) Nucl. Acids Res. 16:5197.
Editor information
Editors and Affiliations
Copyright information
© 1989 Stockton Press
About this chapter
Cite this chapter
Kawasaki, E.S., Wang, A.M. (1989). Detection of Gene Expression. In: Erlich, H.A. (eds) PCR Technology. Palgrave Macmillan, London. https://doi.org/10.1007/978-1-349-20235-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-349-20235-5_8
Publisher Name: Palgrave Macmillan, London
Print ISBN: 978-0-333-48948-2
Online ISBN: 978-1-349-20235-5
eBook Packages: Chemistry and Materials ScienceChemistry and Material Science (R0)