BMCB625:ncRNA: Difference between revisions
(→Jon) |
(→Jon) |
||
| Line 16: | Line 16: | ||
---- | ---- | ||
===Jon=== | ===Jon=== | ||
Q1: Why do you think (in regards to fig 4) that addition of antibody to lane 5 | Q1: Why do you think (in regards to fig 4) that addition of antibody to brain extracts (lane 5) removes three major complexes seen in lane 4? | ||
Revision as of 20:02, 12 June 2007
(Homework) Questions
Larry
Chayne
Jon
Q1: Why do you think (in regards to fig 4) that addition of antibody to brain extracts (lane 5) removes three major complexes seen in lane 4?
Chris
- Q1:
The authors claim that FMRP binding to BC1 RNA is “...specific and stoichiometric since the complex can be easily competed off by 100-fold excess unlabeled BC1 RNA...” (Fig 4, Ln3, versus Ln6&7)
I agree it demonstrates specificity, but how is this stoichiometric?
- Q2:
Figure 6: This is a good assay to demonstrate base-pairing interactions between BC1 and a variety of mRNA’s, but how could we demonstrate this is occurring in vivo?
Mahta
Q1. From the data can you envision a model in which several RNAs similar to BC1 act as "bridges" or "adaptors" between FMRP and mRNAs targetd for degradation?
Q2. High salt is needed to efficiently immunoprecipitate FMRP (the authors state that the C-terminal can be masked at physiological concentrations). Also, the recombinant FMRP is only stable in 1M salt. Is this troublesome?
