SpringerProtocols: Abstract: Use of the GUS Reporter Gene

Use of the GUS Reporter Gene

By: Joy Wilkinson², Keith Lindsey³

Abstract

Full Text

Download PDF (1206K)

One of the most important considerations in the expression of heterologous proteins in plants is the choice of promoter. The study of promoter activity is simplified in the majority of cases by the use of a readily detectable reporter gene. Indeed, reporter genes can also be used to aid the isolation of promoters, through the transformation of plants with a promoterless reporter gene construct. Activation of the reporter gene will then only occur if the gene is inserted adjacent to a native promoter (1).

Affiliation(s): (2) Plant Science, Unilever Research Colworth Laboratory, Bedfordshire, UK
(3) Department of Biological Sciences, University of Durham, UK

Book Title: Recombinant Proteins from Plants: Production and Isolation of Clinically Useful Compounds

Series: Methods in Biotechnology | Volume: 3 | Pub. Date: Dec-24-1997 | Page Range: 39-47 | DOI: 10.1007/978-1-60327-260-5_4

Subject: Plant Sciences

References

Download References

1.	Lindsey, K., Wei, W., Clarke, M. C., McArdle, H. F., Rooke, L. M., and Topping, J. F. (1993) Tagging genomic sequences that direct transgene expression by activation of a promoter trap in plants. Transgenic Res. 2, 33–47.

2.	Herrera-Estrella, L., Depicker, A., Van Montagu, M., and Schell, J. (1983) Expression of chimeric genes transferred into plant cells using a Ti-plasmid-derived vector. Nature 303, 209–213.

3.	An, G. (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol. 81, 86–91.

4.	Jefferson, R. A., Kavanagh, T. A., and Bevan, M. W. (1987) GUS fusions∶ β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901–3907.

5.	Ow, D. W., Wood, K. V., Deluca, M., De Wet, J. R., Helsinki, D. R., and Howell, S. H. (1986) Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234, 856–859.

6.	Niedz, R. P., Sussman, M. R., and Satterlee, J. S. (1995) Green fluorescent protein∶ an in vivo reporter of plant gene expression. Plant Cell Rep. 14, 403–406.

7.	Novel, G. and Novel, M. (1973) Mutants of E. coli Kl2 unable to grow on methyl-β-D-glucuronide∶ map of uid A locus of the structural gene of β-D-glucuronidase. Mol. Gen. Genet. 120, 319–335.

8.	Jefferson, R. A., Burgess, S. M., and Hirsh, D. (1986) β-glucuronidase from E. coli as a gene fusion marker. Proc. Natl. Acad. Sci. USA 83, 8447–8451.

9.	Yang, N-S. and Christou, P. (1990) Cell type specific expression of a CaMV 35S-GUS gene in transgenic soybean plants. Dev. Genet. 11, 289–293.

10.	Terada, R. and Shimamoto, K. (1990) Expression of CaMV 35S-GUS gene in transgenic rice plants. Mol. Gen. Genet. 220, 389–392.

11.	Battraw, M. J. and Hall, T. C. (1990) Histochemical analysis of CaMV 35S pro-moter-β-glucuronidase gene expression in transgenic rice plants. Plant Mol. Biol. 15, 527–538.

12.	Dong, J-Z., Yang, M-Z., Jia, S-R., and Chua, N-H. (1991) Transformation of melon (Cucumis melo L.) and expression from the Cauliflower Mosaic Virus 35S promoter in transgenic melon plants. Bio/Technology 9, 858–863.

13.	Ohta, S., Mita, S., Hattori, T., and Nakamura, K. (1990) Construction and expression in tobacco of a β-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol. 31, 805–813.

14.	Rao, A. G. and Flynn, P. (1990) A quantitative assay for λ-D-glucuronidase (GUS) using microtiter plates. BioTechniques 8, 38–40.

15.	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.

16.	Hu, C-Y., Chee, P. P., Chesney, R. H., Zhou, J. H., Miller, P. D., and O’Brien, W. T. (1990) Intrinsic GUS-like activities in seed plants. Plant Cell Rep. 9, 1–5.

17.	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 Set 70, 133–140.

18.	Wilkinson, J. E., Twell, D., and Lindsey, K. (1994) Methanol does not specifically inhibit endogenous β-glucuronidase (GUS) activity. Plant Sci. 97, 61–67.

Comments (Loading...)

Post comment/View all comments