mRNA stability in E. coli.
rafael at howard.genetics.utah.edu
Tue Jan 16 20:58:57 EST 1996
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.
Rafael Maldonado | La cita ha sido
room 6160 Eccles Institute of Human Genetics |
Department of Human Genetics | retirada por respeto
University of Utah |
Salt Lake City, Utah 84112. USA. | a la propiedad
Rafael.Maldonado at genetics.utah.edu |
Rafael at howard.genetics.utah.edu | intelectual.
Tel: 801-581-4429 |
Fax: 801-585-3910 |
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