In article <l03130302b38ccacd7a84@[129.94.136.176]>,
k.sunn at garvan.unsw.edu.au (Kathryn Sunn) wrote:
> Has anyone ever heard of a cell line which stabley expresses a GFP fusion
> protein, silencing the fluorescence of the GFP?? I have recently made a
> stable cell line with the EGFP vector under the CMV promoter and it was
> working really well, now after approximately 2 months in culture all the 36
> lines of the fusion protein cell expressing cell line no longer fluoresce.
> As you can imagine this is quite a lot of wasted time and it would be
> interesting to know if anyopne else has had the same problem, or knows if
> this promoter is a bit unreliable?
>> Thanks
>> Kate
>> ___________________________________________________________________
> Kathryn Sunn BSc (Hons)
> Bone and Mineral Research Program
> Garvan Institute of Medical Research
> St Vincent's Hospital Tel: (02) 295 8260
> Sydney NSW 2010 Fax: (02) 295 8241
> Australia email: k.sunn at garvan.unsw.edu.au> ___________________________________________________________________
Hi,
I have been able to develop quite a number of stable EGFP fusion protien
expressing cell lines. I was able to maintain expression.. However, once
in a while I sort the cells to increase the average level of expression.
Maybe that could help you. The cell lnes I used were RBL-2H3 and Mc-3T3-E1
cells.
Martin Houle M.Sc.
Centre de Recherche du CHUL
Laval University
Canada
email: martin.houle at crchul.ulaval.ca
--
Martin Houle M.Sc.
Centre de Recherche du CHUL
Laval University, Quebec, Canada
email: martin.houle@(REMOVE)crchul.ulaval.ca
