BACTERIAL GROWTH
A way to count how many bacteria are in a solution
What is plating bacteria, and counting CFUs?
A way to determine where a protein is located in the host cell. (2)
What is creating GFP-fusions (or mNEON fusions), then visualizing host cells?
What is performing subcellular fractionation, then blotting for your protein?
A way to determine if a gene's exression changes under different conditions. (4+)
What is doing a Western blot?
What is creating a reporter construct with the gene's promoter?
What is an ELISA assay?
What is RT-PCR?
A way to determine the effect of a bacterial protein on a bacterial cell.
What is knocking out a bacterial gene?
A way to determine if one protein binds another protein.
What is immunoprecipitating for one protein, then blotting for the other?
A way to determine how well bacteria grow in host cells or tissues. (3)
What is measuring the LD-50 of bacterial needed to kill host cells?
What is mixed infections: (growing bacteria in the host in competition with another bacterial strain)?
What is killing extracellular bacteria with gentomycin, then lysing the eukaryotic cells, plating bacteria, and counting CFUs?
A way to determine the effect of a host protein on a host cell.
What is RNAi?
A way to determine if a protein's expression level affects a phenotype. (2)
What is adding a second copy of the gene on a plasmid?
What is adding a copy of the gene controlled by a strong or inducible promoter?
A way to determine the effect of a bacterial protein on a host.
What is knocking out or mutating a bacterial gene, then infecting host cells with the mutant bacteria?
A way to determine whether different proteins are covalently modified.
What is LCMS?
A way to determine if the host plasma membrane is permeable. (3)
What is adding gentomycin, then lysing eukaryotic cells and growing intracellular bacteria on plates and counting CFUs?
What is adding propidium iodide and determining the level of nuclei staining using flow cytometry?
What is measuring the concentration of LDH that has been released from host cells?
A way to determine if the presence of one protein in a bacteria affects the concentration of another protein.
What is proteomic analysis with LCMS? Compare WT and knockout strains
A way to determine what parts of a protein are necessary for an activity. (2)
What is creating chimeras with a related proteins that don't share that characteristic?
What is creating deletion constructs missing various domains?
A way to determine if a protein exhibits a specific enzymatic activity.
What is mutating the active site, followed by an assay that detects the enzymatic activity?
A way to determine whether endosome acidification is necessary for a process? (2)
What is adding bafilomycin to prevent acidification and determining if the process still happens?
What is measuring the pH of the endosome at the time the process happens?
A way to determine if a protein is degraded by the proteosome. (2)
What is inhibiting new protein synthesis, then measuring whether the protein's concentration decreases over time?
What is IPing the protein and blotting for ubiquitin?
What is a way to determine if one protein's activity in vivo affect another protein's activity?
What is comparing the phenotype of single vs. double knockouts
A way to determine whether a protein binds a DNA sequence?
What is EMSA (electrophoretic mobility shift assay)?
A way to determine if host cells are proliferating?
What is staining tissues w/ Ki67, normalize stain to general DNA stain (DAPI)
A way to determine at which step in a pathway an inducer acts.
What is creating a reporter construct with the promoter from each gene in the pathway?
A way to determine whether a protein binds to chemical X.
What is a variation on an Elisa assay? (attach X to a plastic plate, incubate w/ protein, add fluorescently labeled antibody.)