GFP fusion protein in CHO

[Beat Ludin]

Beatrice Marqueze wrote:

>I have constructed a fusion protein with GPF using Clontech vector
>pEGFP-N1. My goal is to study in CHO cells the distribution of the fusion
>protein in the different sub-cellular compartments.
>
>Does anybody know where GFP alone is distibuted when transfected in CHO, in
>the cytoplasm, in the nucleus ?

Yes, i.e. both in the cytoplasm and in the nucleus with a slight tendency 
to accumulate into the nucleus.

> Could the  fusion protein be wrongly
>localized in the GFP compartments?

We have made a number of GFP fusion proteins so far, and in none of the 
cases we have observed a mislocalisation. I haven't heard of something 
like that happening either. But it's certainly conceivable. For example, 
a protein which normally enters the nucleus could become to big to do so 
because of the increased size of the fusion protein. Or a cytoskeletal 
protein may show even cytoplasmic distribution because the fusion with 
GFP has abolished its binding ability. But all the current evidence I 
know of points against an active GFP-mediated relocalisation. So my 
advice is: go ahead.

Beat



-----------------------------------------------------
Dr. Beat Ludin, FMI, PO Box 2543, 4002 Basel, Switzerland
Tel. +41 61 697 6697 / FAX +41 61 697 3976
Internet:ludin at fmi.ch / Compuserve:100102,1527