Analysis of Proteins in Living Cells - Zaidi

Question 1

How is the GFP gene used?

Answer 1

Acts as a reporter - joins promotor that is active only in special neurons and is used to sense the environment

Question 2

In regards to fluorescence resonance energy transfer (FRET), what does only blue light detection mean?

Answer 2

no protein interaction

Question 3

In regards to fluorescence resonance energy transfer (FRET), what does only green light detection mean?

Answer 3

protein interaction

Question 4

FRAP is a technique where selected regions of cells are bleached and there is visible fluorescence recovery in that region. What is this technique used to study?

Answer 4

monitor the movement of unbleached molecules into the bleached area (example: movement of molecules between golgi and ER)

Question 5

What is photoactivation used to study?

Answer 5

trafficking, turnover, and degradation pathways

Question 6

This is a luminescent protein and is fluorescent only upon binding calcium

Answer 6

Aequorin

note: waves of calcium are formed at the site of sperm entry into the egg. Aequorin emits a flash of light in response to the sudden release of calcium

Question 7

What is total internal reflection fluorescence microscopy (TIRF) used for?

Answer 7

to detect single molecules of proteins

Question 8

How does TIRF work?

Answer 8

laser light shines on cover slip at precise angle; because of total internal reflection, light does not enter the sample and majority of fluorescent molecules not excited; the cell surface that is excited emits light that can be measured