Overexpression of a protien (fwd)
tomasd at bdg10.niddk.nih.gov
Mon Oct 30 14:01:57 EST 1995
In article <46u37a$q4r at sun0.urz.uni-heidelberg.de>, Lars Hesse
<larsh at sun0.urz.uni-heidelberg.de> wrote:
> First of all, try TB medium (Current Protocolls in Molecular Biology)
> instead of LB. LB is not rich enough.
I was using Terrific Broth or Superbroth for all my overexpressions. Then
I by mistake used simple LB, and found no big difference. My feeling is,
that if your construct works, choice of media might slightly improve the
yield. But if the construct does not work, different medium won't make it.
BTW, there was a benchmark article in Biotechniques by Clemens
Suter-Crazzolara and Klaus Unsicker (vol.19, No.2(1995))(sorry, no pages
on my copy) where they used cells scraped off from the plate and put
directly to expression medium with IPTG (instead of growing the cells in
liquid culture prior to induction). They claim they get 1000 fold increase
in yield. I tryed it, but I have got no significant increase in production
of my protein. It might however work better for others.
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