robison at ribo.harvard.edu
Fri Nov 15 13:33:34 EST 1991
ming at ZEUS.VETSCI.WISC.EDU ("Ding Ming") writes:
>........... I was thinking that maybe you can do a 2-dimensional gel
>>electrophoresis of the protein extracted from the cells, and then
>>try to add an RNA oligo which corresponds to the portion of the
>>human RNA component of telomerase that we know ( TTGGGG ?). Since
>>the RNA is associated with the protein in vivo it might bind to it
>>on a gel. Thus if you label the oligo, you might be able to
>>identify the protein blot ??
>> Wael Haddara
> The second dimentional run of 2D PAGE is usually done as SDS-PAGE.
>The gel column which is taken from IEF tube has to be treated under
>denaturation condition before it is loaded. It is hard to beleive
>that the association between the protein and the RNA component still
>exists except the RNA is covalently bond to the protein or the
>association site in the protein is only one or a very few of amino
>acid residues which connected to each other in sequence.
> Ding Ming ~ Telephone: (608)265-2449
> Sweet Protein Section ~ Email: ming at zeus.vetsci.wisc.edu
> Taste Research Lab ~ ming at calshp.cals.wisc.edu
> Department of Vet. Sci. ~ ba209 at cleveland.freenet.edu
> University of Wisconsin-Madison ~ ming at tastelab.vetsci.wisc.edu
It might be possible to attempt this experiment if you first
use UV-induced cross-linking to covalently tether the telomerase to the RNA.
Program in Biochemistry, Molecular, Cellular, and Developmental Biology
robison at nucleus.harvard.edu
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