RSV expression vector needed

Robert Horton horton at molbio.cbs.umn.edu
Thu Apr 22 11:35:33 EST 1993


Dear Netlanders;

I need to find a mammalian transient expression vector for 
use in mouse muscle cells. I am looking for the following 
features:

- 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. 
Thanks.


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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