BACTERIAL GROWTH
A way to determine how many bacteria are in a sample. (2)
What is plating bacteria, and counting CFUs?
What is using a luciferase reporter, measuring luminescence?
A way to determine where a protein or other molecule is located in the host cell. (5)
What is creating GFP-fusions, then visualizing host cells?
What is covalently attaching a fluorophore to a molecule, then visualizing host cells?
What is immunofluorescence (using an antibody and microscopy to detect a protein in a cell)?
What is in situ hybridization?
What is cell fractionation, then doing a Western?
A way to determine if a gene's expression changes under different conditions. (7)
What is a Western blot?
What is a dot blot?
What is creating a reporter construct with the gene's promoter?
What is an ELISA assay?
What is RT-PCR?
What is RNA seq?
What is doing a Northern blot?
A way to determine the effect of a bacterial protein on a host or on the bacteria. (2)
What is knocking out or mutating a bacterial gene?
What is increasing the protein's concentration by adding an extra copy of a gene on a plasmid?
A way to determine if one protein binds another protein. (3 1/2)
What is immunoprecipitating one protein, then blotting for the other?
What is immunoprecipitating one protein, then using mass spec to identify bands?
What is dot blotting?
What is Biacore / surface plasmon resonance (SPR)?
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 a mixed infection (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 if a host protein is necessary for a process. (1)
What is RNAi?
A way to determine if a protein's expression level affects a phenotype. (3)
What is adding a second copy of the gene on a plasmid?
What is adding a copy of the gene controlled by a strong promoter or inducible promoter?
What is RNAi?
A way to determine which parts of a protein are necessary for an activity. (3)
What is creating chimeras with a related proteins that don't share that characteristic?
What is creating deletion constructs missing various domains?
What is creating alanine mutations in the protein?
A way to determine whether a molecule is covalantly modified. (4)
What is LCMS (liquid chromatography / mass spec)?
What is immunoblotting for the modification?
What is a dot blot using an Ab for the modification?
What is IP'ing using an antibody to the modification, then identifying which molecules come down?
A way to determine if bacteria have been internalized into host cells. (3)
What is staining with an Ab, permeabilizing membranes, and staining with a 2nd Ab, then counting how many bacteria are double labeled?
What is adding fluorescent bacteria, then using FACs to count how many host cells are fluorescent?
What is immunofluorescence (fixing host cells, staining with an Ab, visualizing microscopcally)?
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 at which step in a pathway an inducer acts. (1)
What is creating a reporter construct with the promoter from each gene in the pathway?
A way to determine if a protein has a short half life or if its location in the cell changes with time. (1)
What is labeling a protein, then doing a pulse-chase experiment.
A way to determine if a protein exhibits a specific enzymatic activity. (1)
What is mutating the active site, followed by an assay that detects the enzymatic activity?
What is expressing the protein in vitro, then assaying for the enzymatic activity?
A way to determine if aspects of host cell morphology is changed by a bacteria / bacterial protein. (3)
What is staining with DAPI (after adding bacteria or protein)?
What is staining membranes (after adding bacteria or protein)?
What is using immunofluorescence to localizing host cell proteins (after adding bacteria or protein)?
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?
A way to identify which domain of a protein has what function. (3)
What is creating protein chimeras from related proteins with different activities?
What is creating deletion constructs from a protein?
What is mutating amino acids in different regions of a protein?
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?