Green fluorescent protein/β-galactosidase double reporters for visualizingDrosophila gene expression patterns
✍ Scribed by Timmons, Lisa ;Becker, Jodi ;Barthmaier, Peter ;Fyrberg, Christine ;Shearn, Allen ;Fyrberg, Eric
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 345 KB
- Volume
- 20
- Category
- Article
- ISSN
- 0192-253X
No coin nor oath required. For personal study only.
✦ Synopsis
We characterized 120 novel yeast Ga14-targeted enhancer trap lines in Drosophila using upstream activating sequence (UAS) reporter plasmids incorporating newly constructed fusions of Aequorea victoria green fluorescent protein (GFP) and Escherichia coli b-galactosidase genes. Direct comparisons of GFP epifluorescence and b-galactosidase staining revealed that both proteins function comparably to their unconjugated counterparts within a wide variety of Drosophila tissues. Generally, both reporters accumulated in similar patterns within individual lines, but in some tissues, e.g., brain, GFP staining was more reliable than that of b-galactosidase, whereas in other tissues, most notably testes and ovaries, the converse was true. In cases of weak enhancers, we occasionally could detect b-galactosidase staining in the absence of discernible GFP fluorescence. This shortcoming of GFP can, in most cases, be alleviated by using the more efficient S65T GFP derivative. The GFP/b-gal reporter fusion protein facilitated monitoring several aspects of protein accumulation. In particular, the ability to visualize GFP fluorescence enhances recognition of global static and dynamic patterns in live animals, whereas b-galactosidase histochemistry affords sensitive high resolution protein localization. We present a catalog of Ga14-expressing strains that will be useful for investigating several aspects of Drosophila melanogaster cell and developmental biology.