No background plasmid

Dom Spinella dspinella at chugaibio.com
Fri Oct 31 19:32:08 EST 1997


> I've been looking for a plasmid that has a multiple cloning site in the
> middle of a suicide gene, so that an insert is nessesary for the
> transformants to survive. So far the only one that I've seen is one made
> by Invitrogen, which I am told you can't plasmid prep yourself, but have to
> by it from Invitrogen every time you want to use it. 
> 
> Has anyone heard of any other plasmids like this?
> 
> Adam Griffith

I assume you're referring to pZero plasmids from Invitrogen in which the
MCS is in the middle of the lethal ccdB gene.  Maybe I'm missing
something here but can't you purchase some plasmid DNA from them and
then roll your own vector by just cloning any old fragment into the
center of the MCS, preparing a large scale plamid prep from the
recombinant, and then digesting away the insert as needed with the same
enzyme or whatever flanking enzymes you'd like use for your real cloning
work?  Of course, there may be patent and licensing issues, but if you
purchased the plasmid from them initially, who's to know that you made
some more on your own?  Just a thought... -- Dom Spinella



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