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replicating a mammalian expression vector in bacteria

mammalian expression transformation

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3 replies to this topic

#1 ReplicQuestion

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Posted 17 April 2015 - 08:11 AM

Hello,

 

I've received a small amount of plasmid DNA that is a mammalian expression vector. There isn't any bacterial antibiotic selection, only eukaryotic.  Before I conduct the transfection, I want to generate sufficient DNA for repeats. If I were to transform the plasmid into bacteria, would the cells be able to replicate the plasmid DNA for mini/maxiprep?

 

There is an SV40 promotoer, but my understanding is that only eukaryotes can read this.



#2 phage434

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Posted 17 April 2015 - 02:15 PM

I believe it would be highly unusual to have a plasmid which ONLY expresses in a mammalian system. Somehow the plasmid must be made, and almost always that is in E. coli. Do you have sequence for your plasmid? I suspect it has features that allow it to replicate and be selected in coli.



#3 pito

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Posted 20 April 2015 - 10:47 AM

if you have the sequence, post it?

It does indeed seem weird that there is no selection marker for E.coli. almost impossible.


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#4 pavoni.ernesto

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Posted 21 April 2015 - 01:56 AM

Which antibiotic selection (eukaryotic) is present?







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