toxic protein

Rich Dudley rdudley+ at pitt.edu
Fri Jun 11 07:39:57 EST 1999


junz at sfu.ca wrote:

> Hi there,
>
> I have a viral capsid protein gene cloned in pET vector in BL21(DE3)pLysS cells. After 3 hours induction, I compared the lysates before and after induction on PAGE. No difference at all. What's wrong? Is the protein toxic to the host? If it is, how to eliminate?
> Anyway, the final question is how to make the protein express?
>
> Any suggestion are highly appreciated.
>
> Mark

What was the concentration of IPTG you used?  At what OD did you induce?  Did you try purifying from the lysate anyway?  I've had a few 6-His proteins in different vectors that just didn't express highly, but couldn't tell until I tried a purification.  Using large
volumes, I could get a working amount.

rich

--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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