Details
| Original language | English |
|---|---|
| Pages (from-to) | 261-264 |
| Number of pages | 4 |
| Journal | Current Genetics |
| Volume | 17 |
| Issue number | 3 |
| Publication status | Published - Mar 1990 |
| Externally published | Yes |
Abstract
Bacterial β-glucuronidase (GUS) has been described as a useful reporter enzyme for gene fusion studies in bacteria and plants. Here we report the expression of GUS in yeast to illustrate further applications of this enzyme as a quantitative tool for measuring gene activity, as a colour selection marker and as a versatile system for protein targeting studies. There is no intrinsic GUS activity in any yeast strain tested. GUS was expressed in transgenic yeast on a multiple-copy vector under the control of the alcohol dehydrogenase 1 (ADH1) promoter. The enzyme is stable in yeast and its activity may be monitored by very sensitive colorimetric or fluorometric methods in extracts, or by the histochemical reagent 5-bromo-4-chloro-3-indolylglucuronide (X-Gluc) on plates. To test the efficacy of GUS as a reporter for targeting proteins into different subcellular compartments in vivo, we fused the presequence of the mitochondrial tryptophanyl-tRNA-synthetase gene (MSW) to the amino terminus of GUS. The activity of the fusion protein is not substantially impaired and it is imported efficiently into yeast mitochondria.
Keywords
- Chimeric genes, Protein targeting, Reporter gene, Transformation
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
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In: Current Genetics, Vol. 17, No. 3, 03.1990, p. 261-264.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Application of the β-glucuronidase gene fusion system to Saccharomyces cerevisiae
AU - Schmitz, Udo
AU - Lonsdale, David M.
AU - Jefferson, Richard A.
PY - 1990/3
Y1 - 1990/3
N2 - Bacterial β-glucuronidase (GUS) has been described as a useful reporter enzyme for gene fusion studies in bacteria and plants. Here we report the expression of GUS in yeast to illustrate further applications of this enzyme as a quantitative tool for measuring gene activity, as a colour selection marker and as a versatile system for protein targeting studies. There is no intrinsic GUS activity in any yeast strain tested. GUS was expressed in transgenic yeast on a multiple-copy vector under the control of the alcohol dehydrogenase 1 (ADH1) promoter. The enzyme is stable in yeast and its activity may be monitored by very sensitive colorimetric or fluorometric methods in extracts, or by the histochemical reagent 5-bromo-4-chloro-3-indolylglucuronide (X-Gluc) on plates. To test the efficacy of GUS as a reporter for targeting proteins into different subcellular compartments in vivo, we fused the presequence of the mitochondrial tryptophanyl-tRNA-synthetase gene (MSW) to the amino terminus of GUS. The activity of the fusion protein is not substantially impaired and it is imported efficiently into yeast mitochondria.
AB - Bacterial β-glucuronidase (GUS) has been described as a useful reporter enzyme for gene fusion studies in bacteria and plants. Here we report the expression of GUS in yeast to illustrate further applications of this enzyme as a quantitative tool for measuring gene activity, as a colour selection marker and as a versatile system for protein targeting studies. There is no intrinsic GUS activity in any yeast strain tested. GUS was expressed in transgenic yeast on a multiple-copy vector under the control of the alcohol dehydrogenase 1 (ADH1) promoter. The enzyme is stable in yeast and its activity may be monitored by very sensitive colorimetric or fluorometric methods in extracts, or by the histochemical reagent 5-bromo-4-chloro-3-indolylglucuronide (X-Gluc) on plates. To test the efficacy of GUS as a reporter for targeting proteins into different subcellular compartments in vivo, we fused the presequence of the mitochondrial tryptophanyl-tRNA-synthetase gene (MSW) to the amino terminus of GUS. The activity of the fusion protein is not substantially impaired and it is imported efficiently into yeast mitochondria.
KW - Chimeric genes
KW - Protein targeting
KW - Reporter gene
KW - Transformation
UR - http://www.scopus.com/inward/record.url?scp=0025254446&partnerID=8YFLogxK
U2 - 10.1007/BF00312618
DO - 10.1007/BF00312618
M3 - Article
C2 - 2187624
AN - SCOPUS:0025254446
VL - 17
SP - 261
EP - 264
JO - Current Genetics
JF - Current Genetics
SN - 0172-8083
IS - 3
ER -