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Please explain to me - CMVp

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Please explain to me - CMVp

Postby Ammie » Mon Mar 19, 2007 8:30 am

why is it good to have a cytomegalovirus promoter in the cloning plasmid? I do not understand that, please explain to me?
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Postby Jelanen » Tue Mar 20, 2007 1:22 am

So you can have an inducible promoter for expression in eukaryotic cells.
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Postby Ammie » Thu Mar 22, 2007 12:22 pm

Thanks!

But I already have T7, so what is the difference?
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Postby Ammie » Tue Oct 02, 2007 10:56 am

Hi. Thinking about this T7 and CMV promoter again...

I have both in my plasmid.

I have understand that T7 polymeras can be used for in vitro/cell-free trancription. Is that what it is used for?

And CMV promoter as I understood it is good because than you dont need to make a RNA transcript of your gene (in a plasmid) instead you can transfect the whole plasmid when you want to express it in cells.

I still do not really get it. Could anyone explain the differences of those promoters for me or at least give me a hint.
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Postby Duke » Tue Oct 02, 2007 11:37 am

Ammie wrote:Hi. Thinking about this T7 and CMV promoter again...

I have both in my plasmid.

I have understand that T7 polymeras can be used for in vitro/cell-free trancription. Is that what it is used for?

And CMV promoter as I understood it is good because than you dont need to make a RNA transcript of your gene (in a plasmid) instead you can transfect the whole plasmid when you want to express it in cells.

I still do not really get it. Could anyone explain the differences of those promoters for me or at least give me a hint.


T7: Inducible promoter for bacterial expression (e.g. pET system), needs T7 polymerase which is not present in native E.coli but needs to be expressed under the control of a DE3 gene (can be or usually is controlled by the lac-operon)

CMV: constitutive promoter for mammalian expression

Do you have a shuttle vector for both bacterial and mammalian expression?

The CMV promoter on a cloning plasmid is not of any use unless you would like to use the palsmid directly for expression of the protein in mammalian cells.
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