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Class_6_highlights Weber et al

Class_6_highlights Weber et al - Weber et al The SNARE...

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Weber et al The SNARE hypthesis is well accepted for explaining how vesicles identify the proper targets. What hypothesis about SNAREs is being tested in this paper? Name the vSNARE and tSNAREs used in this paper. How was the v-SNARE purified? from bacteria with C-terminal his tag on Ni-agarose beads How were the t-SNAREs purified? from bacteria with N-terminal GST tag on only one of the proteins! Why do they only need a tag on one? How are the liposomes created? Understand that figure 1A shows us the purity of the protein preps and the purity of the liposomes. Why is this very important to the hypothesis? because the goal is to show that the SNARE proteins are SUFFICIENT for fusion. Thus, these proteins better be the ONLY ones present. UNDERSTAND how FRET works to measure fusion. which fluor is being quenched? which is doing the quenching? look at the spectra for NDB and rhodamine and understand why these two fluors are a good match for doing FRET. Where is the key ovelap?
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