blocking transcription in eukariotic cells : how to ?

Matt Thomas mthomas at hsc.usf.edu
Wed Oct 11 13:43:10 EST 2000


> 
> Dear all,        
> 
> We have this cell which is active during its differenciation probably
> through a protein present at its surface. I would like to block
> transcription in this cell to determine the exact time at which the
> protein is transcribed.
> 
> If the transcription is blocked and the cell is still active that
> means that the protein was already present. I am conscious that
> blocking transcription might stop the differenciation and thus
> eventhough the protein might be present and stocked somewhere it won't
> be exported to the cell surface.
> 
> What chemical would you recomand ?
>

I would go with alpha-amanatin.  I know it works really well in vitro and I
suspect it is fine with cells.  When I was in graduate school a person in
the worm lab down the hall wanted to do something similar and I suggested
alpha-amanatin or (damn, can't remember the name, something you typically
put into primer extenshion reactions, actino-myosinD ?).  She did this just
as I was leaving but when I came back to visit a few months later she said
it worked but didn't go into details.  sorry I can't remember more but might
give you a place to start.  I belive Sigma sells both drugs.


-- 
___________________________________________
Matthew Thomas, Ph. D
H. Lee Moffitt Cancer Center, University of South Florida
mthomas at hsc.usf.edu

"I refrain from publishing for fear that disputes and
controversies may be raised against me by ignoramuses."
   Sir Isaac Newton, correspondence to Liebniz






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