Histochemical and Fluorometric Assays for uidA (GUS) Gene Detection
- Magdalena Cervera
- … show all 1 hide
Purchase on Springer.com
$49.95 / €39.95 / £34.95*
* Final gross prices may vary according to local VAT.
Transgenic plant production has been intimately connected to the β-glucuronidase (uidA or GUS) gene used as a reporter marker gene. The enzyme stability and the high sensitivity and amenability of the GUS assay to qualitative (histochemical assay) and to quantitative (fluorometric or spectrophotometric assay) detection are some of the reasons that explain the extensive use of uidA gene in plant genetic transformation. Methods for uidA gene detection have been thoroughly described in the literature. The aim of this chapter is to describe the basic protocols needed for GUS detection in a plant genetic transformation laboratory.
- Novel, G. and Novel, M. (1973) Mutants d’Escherichia coli affectés pour leur croissance sur methyl β-glucuronide: localisation du gene de structure de la β-glucuronidase (uidA). Mol. Gen. Genet. 120, 319–335.
- 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.
- Jefferson, R. A., Bevan, M., and Kavanagh, T. (1987) The use of the Escherichia coli beta-glucuronidase as a gene fusion marker for studies of gene expression in higher plants. Biochem. Soc. Trans. 15, 17–18.
- Hu, C.-Y., Chee, P. P., Chesney, R. H., Zhou, J. H., and Miller, P. D. (1990) Intrinsic GUS-like activities in seed plants. Plant Cell Rep. 9, 1–5. CrossRef
- Kosugi, S., Ohashi, Y., Nakajima, K., and Arai, Y. (1990) An improved assay for β-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous β-glucuronidase activity. Plant Sci. 70, 133–140. CrossRef
- Mascarenhas, J. P. and Hamilton, D. A. (1992) Artifacts in the localization of GUS activity in anthers of petunia transformed with a CaMV 35S-GUS construct. Plant J. 2, 405–408. CrossRef
- Muhitch, M. J. (1998) Characterization of pedicel β-glucuronidase activity in developing maize (Zea mays) kernels. Physiol. Plant. 104, 423–430. CrossRef
- Jefferson, R. A., Burgess, S. M., and Hirsh, D. (1986) β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc. Natl. Acad. Sci. USA 83, 8447–8451 CrossRef
- Thomasset, B., Ménard, M., Boetti, H., Denmat, L. A., Inzé, D., and Thomas, D. (1996) β-Glucuronidase activity in transgenic and non-transgenic tobacco cells: specific elimination of plant inhibitors and minimization of endogenous GUS background. Plant Sci. 113, 209–219. CrossRef
- Serres, R., McCown, B., and Zeldin, E. (1997) Detectable β-glucuronidase activity in transgenic cranberry is affected by endogenous inhibitors and plant development. Plant Cell Rep. 16, 641–646.
- Tör, M., Mantell, S. H., and Ainsworth, C. (1992) Endophytic bacteria expressing β-glucuronidase cause false positives in transformation of Dioscorea species. Plant Cell Rep. 11, 452–456. CrossRef
- Vancanneyt, G., Schmidt, R., O’Connor-Sanchez, A., Willmitzer, L., and Rocha-Sosa, M. (1990) Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacteriummediated plant transformation. Mol. Gen. Genet. 220, 245–250. CrossRef
- Kirchner, G., Kinslow, C. J., Bloom, G. C., and Taylor, D. W. (1993) Non-lethal assay system of β-glucuronidase activity in transgenic tobacco roots. Plant Mol. Biol. Rep. 11, 320–325. CrossRef
- Martin, T., Wöner, R.-V., Hummel, S., Willmitzer, L., and Frommer, W. B. (1992) The GUS reporter system as a tool to study plant gene expression, in GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression (Gallagher, S. R., ed.), Academic Press, San Diego, CA, pp. 23–43.
- Stewart, C. N., Jr. (2001) The utility of green fluorescent protein in transgenic plants. Plant Cell Rep. 20, 376–382. CrossRef
- Gallagher, S. R., ed. (1992) GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression, Academic Press, San Diego, CA.
- 17. Stomp, A.-M. (1992) Histochemical localization of β-glucuronidase, in GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression (Gallagher, S. R., ed.), Academic Press, San Diego, CA, pp. 103–113.
- Wilkinson, J. E., Twell, D., and Lindsey, K. (1994) Methanol does not specifically inhibit endogenous β-glucuronidase (GUS) activity. Plant Sci. 97, 61–67. CrossRef
- Naleway, J. J. (1992) Histochemical, spectrophotometric, and fluorometric GUS substrates, in GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression (Gallagher, S. R., ed.), Academic Press, San Diego, CA, pp. 61–76.
- Guivarc’h, A., Caissard, J. C., Azmi, A., Elmayan, T., Chriqui, D., and Tepfer, M. (1996) In situ detection of expression of the gus reporter gene in transgenic plants: ten years of blue genes. Transgen. Res. 5, 281–288. CrossRef
- Histochemical and Fluorometric Assays for uidA (GUS) Gene Detection
- Book Title
- Transgenic Plants: Methods and Protocols
- Book Part
- pp 203-213
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology™
- Series Volume
- Series ISSN
- Humana Press
- Copyright Holder
- Humana Press
- Additional Links
- Fluorometric GUS detection
- histochemical GUS detection
- reporter marker genes
- uidA gene
- Industry Sectors
To view the rest of this content please follow the download PDF link above.