Abstract
The use of reporter genes in transgenic plants provides an excellent opportunity to investigate the ways in which promoters and other regulatory elements regulate gene expression. Neomycin phosphotransferase II (1), chloramphenicol acetyltransferase (2), luciferase (3), and β-glucuronidase (GUS; 4) genes may each be used to provide some indications of the extent and sites of gene expression. Each reporter gene system has particular requirements for assaying gene expression and distinctive features. The neomycin phosphotransferase and chloramphenicol acetyltransferase systems require the use of radioisotopes or HPLC, whereas the luciferase system requires a luminometer or darkroom facilities. The GUS reporter gene system, in contrast, is quick, easy to use, sensitive, does not require radioisotopes, and is relatively inexpensive. Plant biotechnologists use the Escherichia coli GUSA gene in their assessments of reporter gene activity. The E. coli GUS has a monomeric mol wt of 68 kDa, and exists as a tetramer in vivo (5).
This is a preview of subscription content, log in via an institution.
References
Chen, W.-H., Gartland, K. M. A., Davey, M. R., Sotak, R., Gartland, J. S., Mulligan, B. J., et al. (1987) Transformation of sugarcane protoplasts by direct uptake of a selectable chimaeric gene. Plant Cell Rep. 6, 297–301.
Ward, A., Etessami, P., and Stanley J. (1988) Expression of a bacterial gene in plants mediated by infectious geminivirus DNA. EMBO J. 7, 1583–1587.
de Wet, J. R., Wood, K. V., DeLuca, M., Helinski, D. R., and Subramani S. (1987) Firefly luciferase gene. structure and expression in mammalian cells. Mol. Cell Biol. 7, 725–737.
Jefferson, R. A. (1987) Assaying chimeric gene expression in plants the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405.
Wilson, K. J., Giller, K. E., and Jefferson, R. A. (1991) Beta-Glucuronidase (GUS) operon fusions as a tool for studying plant-microbe interactions, in Advances in Molecular Genetics of Plant-Microbe Interactions (Hennecke, H. and Verma, D. P. S., eds.), Kluwer, The Netherlands, pp. 226–229.
Gallagher, S. R. (ed.) (1992) GUS Protocols, Academic, London.
Jefferson, R. A. (1989) The GUS reporter gene system. Nature 342, 837,838.
Jefferson, R. A., Kavanagh, T. A., and Bevan, M. W. (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901–3907.
Vitha, S., Beneš, K., Michalová, M., and Ondrej, M. (1993) Quantitative β-glucuronidase assay in transgenic plants. Biol. Plant 35, 151–155.
Phillips, J. P., Xing, T., Gartland, J. S., Elliott, M. C., and Gartland, K. M. A. (1992) Variation in β-glucuronidase activity in transgenic sugar beet roots. Plant Growth Reg. 11, 319–325.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Gartland, K.M.A., Phillips, J.P., Vitha, S., Beneš, K. (1995). Fluorometric GUS Analysis for Transformed Plant Material. In: Gartland, K.M.A., Davey, M.R. (eds) Agrobacterium Protocols. Methods in Molecular Biology™, vol 44. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-302-3:195
Download citation
DOI: https://doi.org/10.1385/0-89603-302-3:195
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-302-3
Online ISBN: 978-1-59259-531-0
eBook Packages: Springer Protocols