Hello all luciferase lovers!
I will be happy to share my experience as well, once I have something to
tell. For now, I use RSV-Luc reporter with cloned factor sites
upstream. Co- expression of my activator results in luc levels of 20
fold higher. addition of a putative dominant-negative inhhibitor of the
activator kills this activation, so I think Its a pretty good system.
My half-life question is specific for cell-cycle induction of fos-Luc
construct. That is trickier. If the half-live is really about 4.5
hours, then I may stimulate cells for 6 hours to get over the
"background" a bit.
On 5 Oct 1996, Ferland Louis H. wrote:
> Martin, John, and all the rest of you, Netters,
>> The half-life of the protein is only half of the story. I am in a
> position similar to John's i.e. I want to verify possible (expected)
> rapid inhibition of a transfected promoter driving the LUC reporter. But,
> to have an idea of whether LUC is useable for this type of experiment, we
> need not only the half-life of the LUC protein, but also of its mRNA.
> Does anyone have any data on this?
>> We're just going to give it a try...
>> - Louis
>>> On 2 Oct 1996, martin LEACH wrote:
>> > Date: 2 Oct 1996 20:19:26 GMT
> > From: martin LEACH <leach at bu.edu>
> > To: methods at net.bio.net> > Subject: Re: Luciferase half-life?
> > Hi john
> > I believe the half life of luciferase is around 4.5 hours....(protein)
> > I believe this was determined in NIH fibroblasts...not sure though
> > Martin