Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

conditionally active plasmids - (Jul/07/2013 )

I'm a research student and would like to ask a question regarding the use of conditoonally active plasmid construct. I had read a paper which used HA-FOXO3a.A3-ER construct. This constrcut contains the ER domain that allows the induction of FOXO3a(A3) activity by 4OHT. The authours used this constuct to overexpress FOXO3a in the cells. I was wondering what is the advantge of using this construct over a simple construct expressing only FOXO3a, as they only wanted to activate FOXO3a in the cells?

Paper: Interaction of FOXO with b-catenin inhibits b-atenin/T-cell factor activity. JBC, 2008.



The main advantage is that you are comparing only the effect of adding OHT (and solvent) to the cells so that it is easier to work out if your response is related to the protein of interest, compared to working it out with the transfection, and induction in there. That's assuming that they used the plasmid to make a stable cell line which has the plasmid in all cells.

Another answer may be that, that plasmid is the one they had, so they worked with it, rather than making a new one.