Skip to main content
[Preprint]. 2023 Jul 17:2023.07.17.549367. [Version 1] doi: 10.1101/2023.07.17.549367

Figure 1. Strategy for generating SynLight, a single transgene that expresses both membrane-tagged GFP and mStrawberry-tagged Bruchpilot-Short.

Figure 1.

A, Diagram of an example plasmid containing a UAS vector and codon-optimized 2A peptide coding sequence (Daniels et al., 2014). Flanking either side of 2A are multiple cloning sites and restriction sites that facilitate insertion of two or more genes of interest. B, Diagram of the SynLight plasmid. Using restriction enzymes, the mCD8-GFP coding sequence was inserted preceding the P2A coding sequence and then the Bruchpilot-Short-mStrawberry coding sequence is inserted following the P2A sequence, keeping all sequences in frame. C, Diagram of SynLight mRNA, showing two separate proteins being produced from a single mRNA sequence. D-D”, Representative maximum projection confocal image stacks of multiglomerular LNs of the adult brain expressing SynLight and stained with antibodies against mStraw (red), GFP (green), and N-Cadherin (blue). These images show overlapping, yet distinctly different subcellular localization of Brp-Short-mStraw and mCD8-GFP. E-E”, Representative maximum projection confocal image of the third instar ventral nerve cord expressing SynLight, showing separate endogenous expression of Brp-Short-mStraw and mCD8-GFP via the native fluorescence from the mStrawberry and GFP fluorophores. Scale bars = 40 μm (D); 80 μm (E).