GFP Fusion Protein Expression in Drosophila

Full-length cDNAs for Fzy and Fzr were obtained from Research Genetics and from Christian Lehner (University of Bayreuth, Bayreuth, Germany), respectively. The coding sequences were modified by PCR so that mGFP6 (Schuldt et al., 1998 (link)) could be cloned, in frame, onto the NH₂ terminus of both proteins. The GFP fusion proteins were then subcloned into the pWRpUBq Drosophila transformation vector, putting their expression under the control of the polyubiquitin promoter that is expressed at relatively high levels throughout Drosophila development (Lee et al., 1988 (link)). Full details of these cloning procedures are available upon request. These plasmids were then used to generate stable fly lines using standard P-element–mediated transformation (Roberts, 1986 ).