choice for protein expression system
i.mcfarlane at icrf.icnet.uk
Thu Sep 2 06:21:15 EST 1999
In article <7qg6c0$ssn$1 at fu-berlin.de>, "Oliver Politz"
<politz at medizin.fu-berlin.de> wrote:
> I am about to go for the expression and purification of new protein. So my
> "simple" question is which expression systems are the best choice for this.
> The protein:
> approx. 300 kD incl. glycosylation (important for epitops)
> seems to be type I membrane protein
> can be find in crude cell extracts and to a very small extent in supernatant
> of cell culture
> My ideas
> 1. baculo system (I have a offer from novagen for their system)
> 2. mamalian cell culture transient and/or stable transfected with the
> protein ( it is not expressed in i.e. COS cells)
It really depends on what you want to do with the protein. My preference
would be to make stable transfectants with myc and/or his tags in HeLa or
COS cells. This strategy would give you easy methods to detect/purify the
protein after expression. Another way is make a secreteable construct by
deleting the tm domain, although this may not express well, which will
give higher yields of protein.
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