cotransfect or bicistronic

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Mon Mar 13 11:33:53 EST 2000


This message has been posted by:  612 right <i.hinners at REMOVE-THIS-TO-SENDicrf.icnet.uk>



Soenke Behrends wrote:

> Dear netters,
>
> Is it fair to say that when you transfect pEGFP with green
> fluorescence only and another vector with the same features
> with your gene of interest that the fluorescence leads you
> with some likelihood to the cells expressing you gene of
> interest (e.g. for electrophysiology)?
>
> i have heard (but do not have good scientific reference)
> for the notion that when you transiently transfect cells
> (say with 10% of cells being transfected) you mostly
> get the same 10% of cells cotransfected with the another
> similar vector.

Same for me, Soenke,
people in our lab have the same experience and make use  of this fact. I
dont know of any reference either,
but we find the all cells that are transfected carry both plasmids and
express both proteins in double transfections

Well, not much of help here.... sorry Ina




>
> Fusion protein would of course a good alternative but fluorescence
> is not strong enough in our case.
>
> Or is the use of a bicistronic vector (IRES marker control vector,
> Clontech) a good idea? If the ideas from the first paragraph
> are true I would be happy to get around the cloning involved.
>
> Thanks for your time and help
> Soenke

--
Ina Hinners
ICRF
Secretory Pathways Laboratory
44 Lincolns Inn Fields
WC2A 3PX London
UK
email: I.Hinners at icrf.icnet.uk






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