Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1993 Jun;59(6):1767–1773. doi: 10.1128/aem.59.6.1767-1773.1993

The GUS gene fusion system (Escherichia coli beta-D-glucuronidase gene), a useful tool in studies of root colonization by Fusarium oxysporum.

Y Couteaudier 1, M J Daboussi 1, A Eparvier 1, T Langin 1, J Orcival 1
PMCID: PMC182159  PMID: 8328800

Abstract

The plant-pathogenic fungus Fusarium oxysporum was successfully transformed with the beta-D-glucuronidase gene from Escherichia coli (gusA) (GUS system) in combination with the gene for nitrate reductase (niaD) as the selectable marker. The frequency of cotransformation, as determined by GUS expression on plates containing medium supplemented with 5-bromo-4-chloro-3-indolyl glucuronide (GUS+), was very high (up to 75%). Southern hybridization analyses of GUS+ transformants revealed that single or multiple copies of the gusA gene were integrated into the genomes. High levels of GUS activity are expressed in some transformants, but activity in F. oxysporum does not appear to be correlated with the copy number of the gusA gene. Since the highest activity was found in a transformant with a single copy, it can be assumed that sequence elements of F. oxysporum integrated upstream of the gene can act as a promoter or enhancer. Expression of the gusA gene was also detected during growth of the fungus in plants, indicating that the GUS system can be used as a sensitive and easy reporter gene assay in F. oxysporum.

Full text

PDF
1767

Images in this article

1770Image
on p.1770
1772Image
on p.1772

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. 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. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Bunkers G. J. Expression of the Escherichia coli beta-glucuronidase gene in Pseudocercosporella herpotrichoides. Appl Environ Microbiol. 1991 Oct;57(10):2896–2900. doi: 10.1128/aem.57.10.2896-2900.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cove D. J. The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochim Biophys Acta. 1966 Jan 11;113(1):51–56. doi: 10.1016/s0926-6593(66)80120-0. [DOI] [PubMed] [Google Scholar]
  4. Daboussi M. J., Djeballi A., Gerlinger C., Blaiseau P. L., Bouvier I., Cassan M., Lebrun M. H., Parisot D., Brygoo Y. Transformation of seven species of filamentous fungi using the nitrate reductase gene of Aspergillus nidulans. Curr Genet. 1989 Jun;15(6):453–456. doi: 10.1007/BF00376803. [DOI] [PubMed] [Google Scholar]
  5. Daboussi M. J., Langin T., Brygoo Y. Fot1, a new family of fungal transposable elements. Mol Gen Genet. 1992 Mar;232(1):12–16. doi: 10.1007/BF00299131. [DOI] [PubMed] [Google Scholar]
  6. Jefferson R. A. The GUS reporter gene system. Nature. 1989 Dec 14;342(6251):837–838. doi: 10.1038/342837a0. [DOI] [PubMed] [Google Scholar]
  7. Malardier L., Daboussi M. J., Julien J., Roussel F., Scazzocchio C., Brygoo Y. Cloning of the nitrate reductase gene (niaD) of Aspergillus nidulans and its use for transformation of Fusarium oxysporum. Gene. 1989 May 15;78(1):147–156. doi: 10.1016/0378-1119(89)90322-3. [DOI] [PubMed] [Google Scholar]
  8. Pobjecky N., Rosenberg G. H., Dinter-Gottlieb G., Käufer N. F. Expression of the beta-glucuronidase gene under the control of the CaMV 35s promoter in Schizosaccharomyces pombe. Mol Gen Genet. 1990 Jan;220(2):314–316. doi: 10.1007/BF00260500. [DOI] [PubMed] [Google Scholar]
  9. Punt P. J., Oliver R. P., Dingemanse M. A., Pouwels P. H., van den Hondel C. A. Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene. 1987;56(1):117–124. doi: 10.1016/0378-1119(87)90164-8. [DOI] [PubMed] [Google Scholar]
  10. Roberts I. N., Oliver R. P., Punt P. J., van den Hondel C. A. Expression of the Escherichia coli beta-glucuronidase gene in industrial and phytopathogenic filamentous fungi. Curr Genet. 1989 Mar;15(3):177–180. doi: 10.1007/BF00435503. [DOI] [PubMed] [Google Scholar]
  11. Schmitz U. K., Lonsdale D. M., Jefferson R. A. Application of the beta-glucuronidase gene fusion system to Saccharomyces cerevisiae. Curr Genet. 1990 Mar;17(3):261–264. doi: 10.1007/BF00312618. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES