cotransfect or bicistronic

Mikel Zaratiegui mazarbiu at unav.es
Tue Mar 14 10:46:24 EST 2000


You can think of transfection reagents such as calcium phosphate or
lipid compounds as stuff that coaggregates with DNA. These aggregates
contain up to thousands of copies of the plasmid transfected, and
therefore in a 1:1 mix of two plasmids it is highly unlikely that an
aggregate will contain only one kind of plasmid. So you could say that
if a particular cell has swallowed GFP, it has swallowed the other gene
along with it.
However, if you want to use transient transfection for
electrophysiology, be aware that some transfection reagents modify
membrane structure. Consider a stable transfectant if this is an issue.

Mikel

Soenke Behrends ha escrito:

> 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.
>
> 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





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