Scott_Handout2-key - 2/8/2010 MCB 104; Handout #2 1....

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2/8/2010 MCB 104; Handout #2 1. Anterograde transport: there’s not a ton of memorization in this class, but you should know the order in which a secreted protein passes through the ER, and various golgi cisternae in anterograde transport. Ribosome/Cytoplasm --> ER –[ vesicle ]-> cis golgi –[ cisternal matutation ]-> medial golgi –[ cisternal maturation ]-> trans golgi –[ vesicle ]-> target Anterograde transport is conducted via vesicles; recycles proteins from later compartments back to earlier compartments. 1A. In which steps is the protein transported by vesicle? ER cis golgi; trans golgi target anterograde transport of ER/golgi proteins is also conducted by vesicle 1B. In which steps is the protein transported by cisternal maturation? Cis golgi medial golgi; medial golgi trans gogli 1C. How are proteins that function specifically in the Golgi and ER recycled? anterograde transport 1D. Describe two experimental techniques that you can use to monitor the progression of a protein through anterograde transport in the secretory pathway. 1. Pulse chase with radioactive molecules; arrest the cells at various timepoints, and see which compartments have radioactivity. 2. Express a pulse of GFP-tagged protein. Monitor the progression of the GFP-tagged protein over time. 2A. In the high-throughput screen for secretory pathway mutants in yeast, which cell would have the highest density: wild-type cell at 25C (permissive temp), wild-type cell at 37C (restrictive temp), sec mutant cell at 25C, or sec mutant cell at 37C? Sec mutants at 37C would have the highest density b/c secreted proteins are accumulating
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This note was uploaded on 04/05/2010 for the course MCB 104 taught by Professor Urnov during the Spring '09 term at University of California, Berkeley.

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Scott_Handout2-key - 2/8/2010 MCB 104; Handout #2 1....

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