Exam 9: Tools for Analyzing Gene Expression

A coimmunoprecipitation assay is a process that is used to identify

Which of the following is a key advantage of NMR spectroscopy over X-ray crystallography?

During a transient transfection assay

Describe the use of a vector that produces fusion proteins with glutathione-S-transferase (GST) at one end. Show the protein purification scheme to illustrate the advantage of the GST tag.

The distance moved by a protein into a gel is inversely proportional to the

Compare and contrast the electrophoretic mobility shift assay (EMSA) and DNase I footprinting methods for assaying specific DNA-protein interactions. What information does DNase footprinting provide that EMSA does not?

You have used a polyclonal primary antibody made in rabbit to detect your protein of interest on Western blot. Give an example of an appropriate secondary antibody to use for detection. Explain your choice.

Describe fluorescence in situ hybridization (FISH). When would you use this method, rather than Northern blotting?

You want to generate "optical sections" of a specimen. To this end, would you use conventional fluorescence microscopy or confocal microscopy? Explain your answer.

Compare and contrast transient and stable transfection assays.

You want to confirm the interaction of a particular protein with another protein at a precise location within a cell. What technique would you use and why?

You are testing protein X and protein Y in a yeast two-hybrid assay that uses lacZ as a reporter gene. Production of a blue product signifies that:

Northern blotting is the process that is used to identify

You are interested in an uncharacterized gene that contains an interesting combination of predicted protein domains. To determine in which mouse tissues it is expressed, the best method is

Diagram the key steps in a "sandwich" ELISA. What information can be obtained from this type of assay?

You want to study regulatory regions that control expression of a gene whose protein product is difficult to measure quantitatively. What method should you use to analyze gene expression?

A chromatin immunoprecipitation assay is a process that is used to identify

To map the start of transcription for a given mRNA, the most useful method would be

You want to analyze the structure of a large protein complex at extremely high magnification. What method would you use?

To analyze whether a putative transcriptional enhancer activates transcription, one could use which of the following approaches?