Islating Telomerase

Keith Robison 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
>>
>>
>Dear Wael,

>  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=-

>    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.



Keith Robison
Harvard University
Program in Biochemistry, Molecular, Cellular, and Developmental Biology

robison at nucleus.harvard.edu



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