You have purified protein you call MAP15. MAP 15 binds totubulin that is GTP-bound tubulin and does not bind to tubulin thatis GDP-bound. You genetically engineered cells to express a greenfluorescent protein (GFP) âtagged version of MAP15. This GRP-MAP15fusion protein does not appear to disrupt the function of MAP15.You use a red fluorescent dye to label the microtubules so that youcan visualize GFP-MAP15 and the microtubules at the same time usingfluorescence microscopy (Note that the red dye does not appear toaffect microtubule structure and function).
A. If you were to polymerize some red-labeled microtubules in adish, would you expect GFP-MAP15 to be associated with one or bothends of a microtubule, with the middle of the microtubule, or beevenly distributed throughout the microtubule? Explain.
B. You examine the red microtubules and GFP-MAP15 in a cell ininterphase. Do you think that you will see GFP-MAP15 associatedwith all microtubules? Explain.
C. If you were to take time-lapse images in which you visualizeonly GFP-MAP15 (and not microtubules), what would you expect tosee? Describe what you think would happen to GFP-MAP over time withrespect to its localization inside of this cell.
You have purified protein you call MAP15. MAP 15 binds totubulin that is GTP-bound tubulin and does not bind to tubulin thatis GDP-bound. You genetically engineered cells to express a greenfluorescent protein (GFP) âtagged version of MAP15. This GRP-MAP15fusion protein does not appear to disrupt the function of MAP15.You use a red fluorescent dye to label the microtubules so that youcan visualize GFP-MAP15 and the microtubules at the same time usingfluorescence microscopy (Note that the red dye does not appear toaffect microtubule structure and function).
A. If you were to polymerize some red-labeled microtubules in adish, would you expect GFP-MAP15 to be associated with one or bothends of a microtubule, with the middle of the microtubule, or beevenly distributed throughout the microtubule? Explain.
B. You examine the red microtubules and GFP-MAP15 in a cell ininterphase. Do you think that you will see GFP-MAP15 associatedwith all microtubules? Explain.
C. If you were to take time-lapse images in which you visualizeonly GFP-MAP15 (and not microtubules), what would you expect tosee? Describe what you think would happen to GFP-MAP over time withrespect to its localization inside of this cell.
