Abstract
β-Glucuronidase (GUS) is a very versatile reporter of gene expression that is frequently used in plant molecular biology. The diverse applications of the GUS gene fusion systems (Gallagher, 1992) are based on the detection of the enzymatic activity of GUS in protein extracts or in tissues using fluorometric and histochemical assays respectively (Jefferson, 1987). The histochemical assay has also been used for subcellular localization of GUS fusion proteins, e.g. for the nuclear targeting of important regulatory proteins (for review see Raikehl, 1994). A novel application of the GUS reporter was demonstrated for protein fusions with the A. thaliana ATHSF1 heat shock transcription factor (Lee et al., 1995) using a fluorescence activity staining protocol following gel electrophoresis.
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Abbreviations
- ATHSF1:
-
A. thaliana HSF1
- GUS:
-
glucuronidase
- HS:
-
heat stress (shock)
- HSF:
-
heat shock transcription factor
- MU:
-
methyl umbelliferone
- MUG:
-
4-methyl umbelliferyl glucuronide
- PAGE:
-
Polyacrylamide gel electrophoresis
- RT:
-
room temperature (25 °C)
- SDS:
-
sodium dodecyl sulphate
- uidA:
-
gene encoding GUS
- WT:
-
wild type (untransformed)
- X-Gluc:
-
5-bromo-4-chloro-3-indolyl glucoronide
References
Gallagher SR (1992) GUS protocols: using the GUS gene as a reporter of gene expression. Academic Press, New York.
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Hübel A and Schöffl F (1994) Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein. Plant Mol. Biol. 26: 353–362.
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.
Jefferson RA, Kavanagh TA and Bevan MW (1987) GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.
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Schöffl F, Schröder G, Kliem M and Rieping M (1993) An SAR sequence containing 395 bp DNA fragment mediates enhanced, gene-dosage-correlated expression of a chimaeric heat shock gene in transgenic tobacco plants. Transgenic Res. 2: 93–100.
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© 1997 Kluwer Academic Publishers
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Lee, J.H., Schöffl, F. (1997). GUS activity staining — a powerful tool in plant molecular biology. In: Gelvin, S.B., Schilperoort, R.A. (eds) Plant Molecular Biology Manual. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-5400-0_2
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DOI: https://doi.org/10.1007/978-94-011-5400-0_2
Publisher Name: Springer, Dordrecht
Print ISBN: 978-0-7923-4205-2
Online ISBN: 978-94-011-5400-0
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