• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Dec 20, 1987; 6(13): 3901–3907.
PMCID: PMC553867

GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.

Abstract

We have used the Escherichia coli beta-glucuronidase gene (GUS) as a gene fusion marker for analysis of gene expression in transformed plants. Higher plants tested lack intrinsic beta-glucuronidase activity, thus enhancing the sensitivity with which measurements can be made. We have constructed gene fusions using the cauliflower mosaic virus (CaMV) 35S promoter or the promoter from a gene encoding the small subunit of ribulose bisphosphate carboxylase (rbcS) to direct the expression of beta-glucuronidase in transformed plants. Expression of GUS can be measured accurately using fluorometric assays of very small amounts of transformed plant tissue. Plants expressing GUS are normal, healthy and fertile. GUS is very stable, and tissue extracts continue to show high levels of GUS activity after prolonged storage. Histochemical analysis has been used to demonstrate the localization of gene activity in cells and tissues of transformed plants.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Pfeiffer P, Hohn T. Involvement of reverse transcription in the replication of cauliflower mosaic virus: a detailed model and test of some aspects. Cell. 1983 Jul;33(3):781–789. [PubMed]
  • Reiss B, Sprengel R, Will H, Schaller H. A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984 Oct;30(1-3):211–217. [PubMed]
  • Simpson J, VAN Montagu M, Herrera-Estrella L. Photosynthesis-associated gene families: differences in response to tissue-specific and environmental factors. Science. 1986 Jul 4;233(4759):34–38. [PubMed]
  • Van den Broeck G, Timko MP, Kausch AP, Cashmore AR, Van Montagu M, Herrera-Estrella L. Targeting of a foreign protein to chloroplasts by fusion to the transit peptide from the small subunit of ribulose 1,5-bisphosphate carboxylase. Nature. 313(6001):358–363. [PubMed]
  • Bevan M, Barnes WM, Chilton MD. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucleic Acids Res. 1983 Jan 25;11(2):369–385. [PMC free article] [PubMed]
  • Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • De Greve H, Dhaese P, Seurinck J, Lemmers M, Van Montagu M, Schell J. Nucleotide sequence and transcript map of the Agrobacterium tumefaciens Ti plasmid-encoded octopine synthase gene. J Mol Appl Genet. 1982;1(6):499–511. [PubMed]
  • Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM. Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet. 1982;1(6):561–573. [PubMed]
  • Fraley RT, Rogers SG, Horsch RB, Sanders PR, Flick JS, Adams SP, Bittner ML, Brand LA, Fink CL, Fry JS, et al. Expression of bacterial genes in plant cells. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4803–4807. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. [PubMed]
  • Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. [PMC free article] [PubMed]
  • Guilley H, Dudley RK, Jonard G, Balàzs E, Richards KE. Transcription of Cauliflower mosaic virus DNA: detection of promoter sequences, and characterization of transcripts. Cell. 1982 Oct;30(3):763–773. [PubMed]
  • Herrera-Estrella L, Block MD, Messens E, Hernalsteens JP, Montagu MV, Schell J. Chimeric genes as dominant selectable markers in plant cells. EMBO J. 1983;2(6):987–995. [PMC free article] [PubMed]
  • Holmes DS, Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. [PubMed]
  • Horsch RB, Fraley RT, Rogers SG, Sanders PR, Lloyd A, Hoffmann N. Inheritance of functional foreign genes in plants. Science. 1984 Feb 3;223(4635):496–498. [PubMed]
  • Jefferson RA, Burgess SM, Hirsh D. beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8447–8451. [PMC free article] [PubMed]
  • Jefferson RA, Klass M, Wolf N, Hirsh D. Expression of chimeric genes in Caenorhabditis elegans. J Mol Biol. 1987 Jan 5;193(1):41–46. [PubMed]
  • Jones JD, Dunsmuir P, Bedbrook J. High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J. 1985 Oct;4(10):2411–2418. [PMC free article] [PubMed]
  • Snyder JG, Virta RL, Segreti JM. Evaluation of the phase contrast microscopy method for the detection of fibrous and other elongated mineral particulates by comparison with a STEM technique. Am Ind Hyg Assoc J. 1987 May;48(5):471–477. [PubMed]
  • Kim KS, Shock TL, Goodman RM. Infection of Phaseolus vulgaris by bean golden mosaic virus: ultrastructural aspects. Virology. 1978 Aug;89(1):22–33. [PubMed]
  • Labarca C, Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. [PubMed]
  • Mazur BJ, Chui CF. Sequence of a genomic DNA clone for the small subunit of ribulose bis-phosphate carboxylase-oxygenase from tobacco. Nucleic Acids Res. 1985 Apr 11;13(7):2373–2386. [PMC free article] [PubMed]
  • Novel G, Novel M. Mutants d'Escherichia coli K 12 affectés pour leur croissance sur méthyl-beta-D-glucuronide: localisation of gène de structure de la beta-D-glucuronidase (uid A. Mol Gen Genet. 1973;120(4):319–335. [PubMed]
  • Odell JT, Nagy F, Chua NH. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature. 313(6005):810–812. [PubMed]
  • Otten LA, Schilperoort RA. A rapid micro scale method for the detection of lysopine and nopaline dehydrogenase activities. Biochim Biophys Acta. 1978 Dec 8;527(2):497–500. [PubMed]
  • Ow DW, DE Wet JR, Helinski DR, Howell SH, Wood KV, Deluca M. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science. 1986 Nov 14;234(4778):856–859. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...