drosophila closures genesee scientific  (Genesee Scientific)

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

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: Drosophila media

Article Snippet: Flugs - plastic fly bottles, Drosophila closures , Genesee Scientific , Cat# 49-100.

Techniques: Concentration Assay

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:

Article Snippet: Flugs - plastic fly bottles, Drosophila closures , Genesee Scientific , Cat# 49-100.

Techniques: Recombinant, Protease Inhibitor, Magnetic Beads, Bicinchoninic Acid Protein Assay, Software, Mass Spectrometry

Journal: STAR Protocols

Article Title: A protocol to use Drosophila melanogaster larvae to model human glioblastoma

doi: 10.1016/j.xpro.2022.101609

Figure Lengend Snippet: Genetic crossing scheme to create a Drosophila EGFR-PI3K GBM model Genetic crossing scheme used to derive the stable experimental stock described in the protocol. TM3 Sb and TM6B Tb Gal 80 are balancer chromosomes and markers used to select progeny of the desired genotype. repo-Gal4 is expressed in a glial specific manner throughout larval life. UAS-dEGFR λ UAS-dp110 CAAX are transgene constructs are expressed by repo-Gal4 to drive GBM-like glial neoplasia. UAS-CD8-GFP provides a glial specific GFP label for neoplastic glia.

Article Snippet: Flugs® - Wide Plastic Vials, Drosophila Closures [AS275] , Genesee Scientific , Cat #49-101.

Techniques: Construct

Journal: STAR Protocols

Article Title: A protocol to use Drosophila melanogaster larvae to model human glioblastoma

doi: 10.1016/j.xpro.2022.101609

Figure Lengend Snippet:

Article Snippet: Flugs® - Wide Plastic Vials, Drosophila Closures [AS275] , Genesee Scientific , Cat #49-101.

Techniques: Recombinant, Saline, Electron Microscopy, Plasmid Preparation, Software, Spot Test, Microscopy

Journal: STAR Protocols

Article Title: A protocol to use Drosophila melanogaster larvae to model human glioblastoma

doi: 10.1016/j.xpro.2022.101609

Figure Lengend Snippet: Drosophila Cornmeal Medium

Article Snippet: Flugs® - Wide Plastic Vials, Drosophila Closures [AS275] , Genesee Scientific , Cat #49-101.

Techniques: Concentration Assay

Journal: Applied and Environmental Microbiology

Article Title: A Putative Acetylation System in Vibrio cholerae Modulates Virulence in Arthropod Hosts

doi: 10.1128/AEM.01113-18

Figure Lengend Snippet: Introduction of the complete deletion of yfiQ restores both virulence in Drosophila and growth on acetate minimal medium to the V. cholerae ΔcobB strain. (A) Deletion of yfiQ does not impact growth on minimal medium supplemented with acetate (10 mM), with growth being similar to that of the wild-type strain. Introduction of the ΔyfiQ mutation restores growth to the strain carrying the deletion in ΔcobB, as both the ΔyfiQ→ΔcobB and the ΔcobB→ΔyfiQ strains grow similarly to the wild-type strain in acetate minimal medium. (B) In LB broth, all strains grow similarly to the wild type. (C) YfiQ does not significantly alter virulence of Drosophila (P > 0.05). In seven independent trials, deletion of yfiQ significantly increased virulence in one assay (P = 0.0003), reduced virulence in a second assay (P = 0.0143), and did not alter virulence in the remaining five assays, including the representative assay presented here. When infected with the ΔyfiQ→ΔcobB strain, fly mortality is increased significantly relative to the strain carrying the ΔcobB deletion alone (P < 0.0001 by a log rank test). This result was reproducible in five independent trials. (D) When flies were infected with the ΔcobB→ΔyfiQ strain, virulence was similarly restored relative to the strain carrying the single cobB deletion (P < 0.0001 by a log rank test).

Article Snippet: V. cholerae strains were streaked for single colonies from glycerol stocks, grown overnight in LB broth, and then diluted 1:10 and added to a cellulose acetate Drosophila vial closure (Genesee Scientific) at the bottom of a standard fly vial.

Techniques: Mutagenesis, Infection

Journal: Applied and Environmental Microbiology

Article Title: A Putative Acetylation System in Vibrio cholerae Modulates Virulence in Arthropod Hosts

doi: 10.1128/AEM.01113-18

Figure Lengend Snippet: Deletion of V. cholerae SIO cobB significantly improves survival of Drosophila without appreciably affecting colonization. (A) Survival of flies fed the V. cholerae wild-type SIO, SIO ΔcobB, or SIO Δacs strain, or uninoculated LB broth as a control, was monitored over 143 h. This assay was performed with V. cholerae strains added to triplicate vials, with 8 to 10 flies per vial. This result is representative of data from five separate trials. Flies fed the SIO ΔcobB or SIO Δacs strain survived significantly longer than did flies fed the wild-type strain (P < 0.0001 by a log rank test). (B) Colonization of flies infected with the V. cholerae SIO wild-type, SIO ΔcobB, or SIO Δacs strain. The assay was performed in triplicate vials, and all surviving flies were collected after 24 and 48 h of infection, homogenized, and plated on selective medium. The bacterial loads of flies infected with the mutant strains were not significantly different from bacterial loads of those infected with the wild-type strain (P > 0.05 by a Mann-Whitney test).

Article Snippet: V. cholerae strains were streaked for single colonies from glycerol stocks, grown overnight in LB broth, and then diluted 1:10 and added to a cellulose acetate Drosophila vial closure (Genesee Scientific) at the bottom of a standard fly vial.

Techniques: Control, Infection, Mutagenesis, MANN-WHITNEY

Journal: Applied and Environmental Microbiology

Article Title: A Putative Acetylation System in Vibrio cholerae Modulates Virulence in Arthropod Hosts

doi: 10.1128/AEM.01113-18

Figure Lengend Snippet: Deletion of the putative deacetylase AcuC does not alter virulence in Drosophila. (A) Survival of flies fed the wild-type, ΔyfiQ, ΔacuC, ΔacuC ΔyfiQ, or ΔacuC ΔyfiQ ΔcobB strain. Deletion of acuC did not alter fly survival relative to the wild type in each of two separate assays, one of which is represented here (P > 0.05 by a log rank test). (B) Survival of flies fed the wild-type, ΔcobB, ΔacuC, or ΔacuC ΔcobB strain. Deletion of acuC in the ΔcobB strain did not alter survival relative to the ΔcobB deletion alone in each of two separate assays (P > 0.05 by a log rank test).

Article Snippet: V. cholerae strains were streaked for single colonies from glycerol stocks, grown overnight in LB broth, and then diluted 1:10 and added to a cellulose acetate Drosophila vial closure (Genesee Scientific) at the bottom of a standard fly vial.

Techniques: Histone Deacetylase Assay

Journal: Applied and Environmental Microbiology

Article Title: A Putative Acetylation System in Vibrio cholerae Modulates Virulence in Arthropod Hosts

doi: 10.1128/AEM.01113-18

Figure Lengend Snippet: CobB alters accumulation of triglycerides in the Drosophila gastrointestinal tract. (A to D) Representative images of Drosophila GI tracts infected with the V. cholerae wild-type (A), Δacs (B), or ΔcobB (D) strain or provided sterile LB broth (D); visualized by confocal microscopy; and stained with DAPI and BODIPY. Bar, 10 μm. (E) Quantification of triglyceride puncta stained with BODIPY 493/503 in midguts of Drosophila flies infected with the V. cholerae wild-type, Δacs, or ΔcobB strain or sterile LB broth, with the levels of significance indicated (****, P < 0.0001; **, P = 0.0060; *, P = 0.0127). These results were representative of data from three trials comparing the wild-type and ΔcobB strains, two of which included the Δacs strain.

Article Snippet: V. cholerae strains were streaked for single colonies from glycerol stocks, grown overnight in LB broth, and then diluted 1:10 and added to a cellulose acetate Drosophila vial closure (Genesee Scientific) at the bottom of a standard fly vial.

Techniques: Infection, Sterility, Confocal Microscopy, Staining