mRNA stability in E. coli.

[Rafael Maldonado]

Hello netters:

I am trying to test the stability of some mRNA from plasmids in E. coli. 

Since they code for protein fussions with LacZ, I was trying the old method 
based on lacZ activity of Kepes (BBA 76, 293). Mainly, it consits of a 
short induction with IPTG (I have a tac promoter), wash, and mesure the 
falling of lacZ activity during the time. 

Instead, my activity increases. I am also trying to 
inhibit transcription after the induction using rifampicin, 
translation using chloranphenicol. It does not seem to work.

Do you have some kind of protocol for this experiments? Papers don't help 
too much. Any other suggestion about how to test that (I know the northern 
and primer extension possibilities) would be appreciated.



Rafa 


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                                             |
Rafael Maldonado                             |  La cita ha sido
room 6160 Eccles Institute of Human Genetics |  
Department of Human Genetics		     |  retirada por respeto
University of Utah			     |
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Rafael.Maldonado at genetics.utah.edu           |
Rafael at howard.genetics.utah.edu	             |  intelectual.
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