Please explain to me - CMVp

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
User avatar
Ammie
Garter
Garter
Posts: 44
Joined: Wed Aug 16, 2006 6:48 am

Please explain to me - CMVp

Post by 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?

Jelanen
Inland Taipan
Inland Taipan
Posts: 218
Joined: Fri Mar 04, 2005 1:00 am
Location: The Lab
Contact:

Post by Jelanen » Tue Mar 20, 2007 1:22 am

So you can have an inducible promoter for expression in eukaryotic cells.
'It is futile to pretend to the public that we understand how an amoeba evolved into a man, when we cannot tell our students how a human egg produces a skin cell or a brain cell!'

Dr Jérôme J. Lejeune

User avatar
Ammie
Garter
Garter
Posts: 44
Joined: Wed Aug 16, 2006 6:48 am

Post by Ammie » Thu Mar 22, 2007 12:22 pm

Thanks!

But I already have T7, so what is the difference?

User avatar
Ammie
Garter
Garter
Posts: 44
Joined: Wed Aug 16, 2006 6:48 am

Post by 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.

Duke
Garter
Garter
Posts: 4
Joined: Wed Sep 26, 2007 1:41 pm

Post by 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.

Post Reply

Who is online

Users browsing this forum: No registered users and 3 guests