Yeast Protocol pp 257-264 | Cite as

Study of Transcriptional Regulation Using a Reporter Gene Assay

  • Yu Fu
  • Wei Xiao
Part of the Methods in Molecular Biology book series (MIMB, volume 313)


Study of gene expression can be facilitated by using a reporter gene assay. Instead of directly measuring the level of target gene mRNA, one can clone the promoter region of the gene of interest in front of a reporter gene and measure the reporter gene expression as a reflection of the expression of the gene of interest. We describe a simple lacZ-fusion system to measure the activity of the reporter gene product β-galactosidase. Different strategies of making the fusion construct and their applications are also discussed. This method is particularly useful to dissect the promoter region of the gene of interest and is also used in other experimental protocols such as the yeast two-hybrid analysis.


Reporter Gene Assay Translational Fusion Multicopy Plasmid Upstream Activate Sequence Transform Yeast Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Guarente, L. and Ptashne, M. (1981) Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 78, 2199–2203.PubMedCrossRefGoogle Scholar
  2. 2.
    Mount, R. C., Jordan, B. E., and Hadfield, C. (1996) Reporter gene systems for assaying gene expression in yeast, in Yeast Protocols: Methods in Cell and Molecular Biology, vol. 53, (Evans, I., ed.). Humana Press, Totowa, NJ, pp. 239–248.CrossRefGoogle Scholar
  3. 3.
    Guarente, L. (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 101, 181–191.PubMedCrossRefGoogle Scholar
  4. 4.
    Rose, M., Casadaban, M. J., and Botstein, D. (1981) Yeast genes fused to β-galactosidase in Escherichia coli can be expressed normally in yeast. Proc. Natl. Acad. Sci. USA 78, 2460–2464.PubMedCrossRefGoogle Scholar
  5. 5.
    Jia, X., Zhu, Y., and Xiao, W. (2002) A stable and sensitive genotoxic testing system based on DNA damage induced gene expression in Saccharomyces cerevisiae. Mutat. Res. 519, 83–92.PubMedCrossRefGoogle Scholar
  6. 6.
    Bartel, P. L. and Fields, S. (1995) Analyzing protein-protein interactions using two-hybrid system. Methods Enzymol. 254, 241–263.PubMedCrossRefGoogle Scholar
  7. 7.
    Xiao, W., Singh, K. K., Chen, B., and Samson, L. (1993) A common element involved in transcriptional regulation of two DNA alkylation repair genes (MAG and MGT1) of Saccharomyces cerevisiae. Mol. Cell. Biol. 13, 7213–7221.PubMedGoogle Scholar
  8. 8.
    Liu, Y. and Xiao, W. (1997) Bidirectional regulation of two DNA-damage-inducible genes, MAG1 and DDI1, from Saccharomyces cerevisiae. Mol. Microbiol. 23, 777–789.PubMedCrossRefGoogle Scholar
  9. 9.
    Myers, A. M., Tzagoloff, A., Kinney, D. M., and Lusty, C. J. (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene 45, 299–310.PubMedCrossRefGoogle Scholar
  10. 10.
    Sherman, F., Fink, G. R., and Hicks, J. (1983) Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  11. 11.
    Ito, H., Fukuda, Y., Murata, K., and Kimura, A. (1983) Transformation of intact yeast cells treated with alkali cations. J. Bacteriol. 153, 163–168.PubMedGoogle Scholar
  12. 12.
    Parent, S. A., Fenimore, C. M., and Bostian, K. A. (1985) Vector systems for the expression, analysis and cloning of DNA sequences in S. cerevisiae. Yeast 1, 83–138.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2006

Authors and Affiliations

  • Yu Fu
    • 1
  • Wei Xiao
    • 1
  1. 1.Department of Microbiology and ImmunologyUniversity of SaskatchewanSaskatoonCanada

Personalised recommendations