SV-40 Promoter Bashing

Joe M. SandyFaulkner, Jr. sfaulkne at compdna.win.net
Wed Aug 17 11:07:07 EST 1994


We are looking for a minimal promoter to use in eucaryotic
expression (transfection) experiments.  Because of viral packaging
constraints, and a very large gene, we need a functional promoter
smaller than the usual SV40 or other currently available
promoters.  We are looking for something smaller than 400 bp, but
preferably in the range of 200 bp.  Has anyone done (and
published) promter bashing on SV40 to shave off previously
identified promoter sequence to identify more minimal yet functional
promoter elements?  For that matter, is such a smaller eucaryotic
promoter commercially available in current-generation expression
constructs? (its been a LONG time since we've done this; ~10
years!) Also, are other entirely different promoters in vogue for
eucaryotic transfection?  We are interested again in something
small (<400 bp) necessary and sufficient for expression in
any mammalian (murine and/or human) cell lines (primary culture,
or established/immortal) of the following types: liver,
epithelial, endothelial, kidney, tracheal, fibroblast.  We know that
the definition of expression depends somewhat on the sensitivity
for detection of the reporter gene, and of course we would be
interested in the strongest possible expression, as we don't have
the most sensitive assay. If anyone has a lead on this, we would
appreciate it. 

David Malehorn
17 August 1994 09:44 CDT





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