Journal: STAR Protocols
Article Title: Protocol for affinity purification-mass spectrometry interactome profiling in larvae of Drosophila melanogaster
doi: 10.1016/j.xpro.2024.103064
Figure Lengend Snippet: Strategy to determine the solubility of a tagged bait protein (A) Schematic illustration of the binary GAL4/UAS expression system. One fly line contains the GAL4 protein under the control of the Mef2 promoter. The other fly stock possesses a UAS-based transgene containing the FLAG-tagged Strip protein. Mating of these two lines will produce progeny that express the GAL4 protein and the UAS element within the same cells. In this study, FLAG::Strip will be made in all cells that normally express the Mef2 protein. (B) Representation of the Drosophila pUAST transformation vector that was used to generate transgenic flies expressing FLAG::Strip. Note the vector also contains an ampicillin resistance gene for growth in E. coli and a white+ (w+) gene for the selection of transformants by eye color. (C) Western blot validating the expression and solubility of the FLAG::Strip protein in larval muscle tissue ( Mef2>FLAG::Strip ). Mef2>lacZ was used as a negative control.
Article Snippet: Flugs - plastic fly bottles, Drosophila closures , Genesee Scientific , Cat# 49-100.
Techniques: Solubility, Expressing, Control, Stripping Membranes, Transformation Assay, Plasmid Preparation, Transgenic Assay, Selection, Western Blot, Negative Control