GFP and DsRed?

Disterer, Petra (Medsch Hampstead/Medicine) p.disterer at medsch.ucl.ac.uk
Thu Aug 3 13:32:05 EDT 2006


Dear list members,

 

 

I am currently interested in expressing two fluorescent proteins from
one plasmid which will have an IRES sequence between the two proteins.
For reasons which would take too long to explain, these fluorescent
proteins need to be from a different species (sequence has to be
significantly different). 

 

I would like to evaluate the expression of both proteins on a FACS
machine. We have a FACSCalibur and a Vantage available. Both have the
488nm and a 635nm laser and the Vantage a 408nm UV source. The
percentage of cells expressing both proteins would be >1%.

 

I'm currently thinking about using GFP and DsRed2 and the 488nm
excitation for both; however I'm not sure this would be the best
solution. I've found this reference saying it can be done (Hawley 2004
Multiparameter flow cytometry of fluorescent protein reporters), but
messages from other websites/boards have repeatedly said that there are
difficulties in detecting the DsRed. Therefore I would like to get some
advice from people who have done co-expression of the two proteins.
Alternatively, if anyone can think of a better combination of
fluorescent proteins (from different species), I'd love to hear it.

 

Thanks a lot for any help

 

Petra Disterer

The UCL Inst. Of Hepatology

London,UK

 

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