nonEGFP fluorescent markers

Mitch Dushay dushay at UMBI.UMD.EDU
Fri Dec 4 10:00:22 EST 1998


Several fluorescent proteins besides EGFP are now offered by Clontech.  We
wanted a second marker, so we ordered the first one available:  EBFP.  We
put the EBFP gene behind the Actin 5C promoter and transfected mosquito cells.
Unfortunately, we did not see marked fluorescence above background with
Actin5C-EBFP, while matched Actin5C-EGFP controls showed strong fluorescence -
we used the appropriate filter sets for each, so it was not a technical
problem.
(EBFP-N1; the plasmid with a CMV promoter from Clontech, didn't show strong
fluorescence either, and Clontech could not tell us whether the CMV promoter
works in insect cells.)
   We bit the bullet and ordered ECFP and EYFP.  I have not yet fused
either to the Actin promoter, but coli carrying plasmids with the genes
behind the lac promoter are noticeably blue and yellow under room light.
We won't know how they work in flies or mosquitoes for a little while, but
the coli results are promising.  So with that disclaimer, if you want a
fluorescent tag other than EGFP, ECFP or EYFP are probably better bets
than EBFP.

Mitch Dushay          dushay at umbi.umd.edu      (301)405-7681 or 405-5111
UMBI, Center for Agricultural Biotechnology
5115 Plant Sciences Building
College Park, MD  20742-4450





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