Green fluorescent protein as a convenient and versatile marker for studies on functional genomics in Drosophila.
Gene function can be deduced from lack or gain of activity. For the manipulation of gene doses or activity in Drosophila, a set of P-based vectors was constructed containing green fluorescent protein as marker. pBLUEi, pGREENi and pYELLi were designed for large insert transformation. Mosaicism was generated in vivo with pFlipG which is also ideal for targeted gene disruption. Tissue-specific gene silencing in vivo was performed with the vector set pHIBS and pUdsGFP. pHIBS allows easy cloning and shuttling of double-headed constructs. With pUdsGFP, double stranded RNA can be produced in defined patterns and the area of interference simultaneously visualized by green fluorescence. We demonstrate nearly complete silencing of a ubiquitously expressed gene in a tissue-specific manner.
Nagel
Anja C
AC
Universität Hohenheim, Institut für Genetik (240), Garbenstrasse 30, 70599 Stuttgart, Germany.
Maier
Dieter
D
Preiss
Anette
A
D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology; D017930:Genes, Reporter; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D009928:Organ Specificity; D012330:RNA, Double-Stranded
D005819:Genetic Markers; D008164:Luminescent Proteins; D012330:RNA, Double-Stranded; D049452:Green Fluorescent Proteins