RSV expression vector needed
horton at molbio.cbs.umn.edu
Thu Apr 22 11:35:33 EST 1993
I need to find a mammalian transient expression vector for
use in mouse muscle cells. I am looking for the following
- RSV promoter
- high copy number plasmid
- transcription termination site
- intron that gets spliced out of the vector-encoded sequence
- NO other genes expressed in mammalian cells
I have looked through all of the catalogs I could get my
hands on, and haven't found a vector that completely fits the
bill. Many of the "modern" vectors have fantastic additions
such as EBNA replication proteins etc. so that they will be
propagated extrachromasomally, but I would rather not have
these parts as I don't want to express these other proteins.
Some of the older plasmids are nice and simple, but they are
pBR322 based, and I need to grow a lot of plasmid (yeah I
know, its just peebeearophobia, but I really like pUC
plasmids better!). Many of the simple expression plasmids
also contain a CMV promoter. I know this will work in muscle
cells, but it is my understanding that RSV works better (any
input on this would be appreciated; I can't really document
that RSV > CMV in muscle cells, it's just what I've heard).
A lot of expression vectors also contain introns, because
getting spliced supposedly has a beneficial effect on gene
expression (I guess). As far as I'm concerned, this seems
like a great idea because it will allow me to monitor
expression by PCR to look for spliced recombinant messages.
I would greatly appreciate it if anyone can help me out.
If you have my dream vector, or know where I can get or buy
it, please drop me a note. Also, any info regarding transient
expression of genes in mouse muscle cells would be great.
Bob Horton /\ "Crash programs fail because of the theory that
U. of Minnesota, CBS || with nine women pregnant you get a baby a month"
1479 Gortner Ave. /||\ -Werner von Braun. Disclaimer:"Bob who?"
St. Paul, MN 55108 ^^ horton at molbio.cbs.umn.edu/(612) 624-3790
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