9 what caveats arises from creating fusions of gfp

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9. What caveat(s) arises from creating fusions of GFP with the carboxy terminus of cellular proteins? GFP is an approximately 27 kDa protein and was fused to the carboxy terminus of all proteins in the yeast genome. Not all proteins behave as wild type when a large bulky fusion added on. This may explain why some 2000 protein fusions did not have fluorescence signal above background.
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Application: Go the web site that provides images of every GFP fusion from the above study: Feel free to explore the site as you answer the following questions: 10. POL1 is the gene that encodes for DNA polymerase. Examine the localization of the GFP fusion to this gene product. Does its localization agree with your idea of the function of POL1? Why? yes, because its in the nucleus where you would expect it to be 11. MIP1 is also a DNA polymerase. Where does it localize? Mitochondria 12. Propose a hypothesis as to why MIP1 is localized to where it is. MIP1 localizes to the mitochondria. This suggests that the polymerase is involved in replicating DNA in the mitochondria. Since mitochondria contain their own genome, it is reasonable to hypothesize that it is required to maintain the mitochondrial genome. 13. Where does Sec61 localize? ER 14. Where does Sec33 localize? Golgi
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