You have purified protein you call MAP15. MAP 15 binds to tubulin that is GTP-bo

Question

You have purified protein you call MAP15. MAP 15 binds to tubulin that is GTP-bound tubulin and does not bind to tubulin that is GDP-bound. You genetically engineered cells to express a green fluorescent protein (GFP) –tagged version of MAP15. This GRP-MAP15 fusion protein does not appear to disrupt the function of MAP15. You use a red fluorescent dye to label the microtubules so that you can visualize GFP-MAP15 and the microtubules at the same time using fluorescence microscopy (Note that the red dye does not appear to affect microtubule structure and function).

A. If you were to polymerize some red-labeled microtubules in a dish, would you expect GFP-MAP15 to be associated with one or both ends of a microtubule, with the middle of the microtubule, or be evenly distributed throughout the microtubule? Explain.

B. You examine the red microtubules and GFP-MAP15 in a cell in interphase. Do you think that you will see GFP-MAP15 associated with all microtubules? Explain.

C. If you were to take time-lapse images in which you visualize only GFP-MAP15 (and not microtubules), what would you expect to see? Describe what you think would happen to GFP-MAP over time with respect to its localization inside of this cell.

Explanation / Answer

A.    If you were to polymerize some red-labeled microtubules in a dish, would you expect GFP-MAP15 to be associated with one or both ends of a microtubule, with the middle of the microtubule, or be evenly distributed throughout the microtubule? Explain.

Answer: Here GFP is a tagging protein with MAP15 and it binds with tubulin only, as tubulin are present at the ends, so GFP_MAP15 will be associated at the ends only.

B.    You examine the red microtubules and GFP-MAP15 in a cell in interphase. Do you think that you will see GFP-MAP15 associated with all microtubules? Explain.

Answer: Interphase is preparatory phase for cell division which is divided into three main phases G1-S-G2. G1 is the phase where maximum protein expresses and as cell is ready for division, GFP-MAP15 will also be associated with all microtubules.

C.   If you were to take time-lapse images in which you visualize only GFP-MAP15 (and not microtubules), what would you expect to see? Describe what you think would happen to GFP-MAP over time with respect to its localization inside of this cell.

Answer: GFP-MAP15 on visualization will give green colour due to Green fluorescent protein and its bindin with MAP15. Overtime as this complex will bind with microtubules and insite the cell it will be gradually appear/move at the ends near centriole region inside the cell. And with labeling of red colur we can see the localization easily at the ends.

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