Searching for CMV mammalian expression vector
btaylor at muskwa.ucs.ualberta.ca
Fri Jul 16 00:34:49 EST 1993
Hi. I am a PhD student in the field of malaria, and my new
focus has become using "naked DNA" immunizations to elicit
specific antibodies to certain malaria proteins.
Already I've attempted a few experiments using a plasmid with
the SV40 SRalpha promoter (rat animal model) with little success
even after three boosts. I've injected plasmids with genes encoding
small secreted proteins, and also plasmids with larger, glycosylated
cell-surface proteins. The small secreted proteins seem to give
the best results, but the immune response is still poor.
My next goal is to try using a plasmid with a different promoter--
namely the CMV promoter. After doing a bit of reading and talking
with friends I've come up with an idea of some of the different
features this plasmid should have:
hCMV IE1 promoter + intron A high level of expression
polylinker Xba, Not, BamH1, etc.
SV40 ori, polyoma ori
PolyA adenylation signal
Amp resistance marker
I have all of these components in one form or another, I'm just going
to have to do some PCRing, cutting and pasting to get them all together.
Does anyone know if this kind of vector has already been constructed,
and if so, how I might be able to get some? I don't mind taking a
little time out to make a vector, but perhaps its already been done.
Also, if anyone has any comments about "naked DNA" immunizations and
how they actually work, I'd like to hear from you.
Would adjuvants somehow help in eliciting a response?
Why do secreted proteins elicit a stronger response than membrane proteins?
And I have many, many more questions....
Thanks for your time,
Dept. of Medical Microbiology and Infectious Diseases
University of Alberta CANADA
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