Expression of a 300kDa protein possible?

Richard P. Grant rgrant at netscape.net
Mon Apr 3 02:43:30 EST 2000


In article <8c9djd$2jpi$1 at gwdu67.gwdg.de>, Christina Philipp 
<philipp at mpiz-koeln.mpg.de> wrote:

> Hello Ng,
> 
> until now, I tried two methods to express my protein:
> 1. TNT T7 Quick Coupled Transcription/Translation System (Promega).
> The luciferase control worked well, but I did not see my desired
> protein.
> 2. A "normal" expression (T7 promoter, induction with 1mM IPTG) in E.
> coli HMS174(DE3).
> 
> Has anyone ever had success with the expression of such a large
> protein?


I successfully expressed a 220 kDa protein once, it required 1 % glucose 
and 1 mM Mg2+ adding to the 2xTY (I kept getting premature termination 
of translation) . . .  it may help you.

-- 
Richard P. Grant MAD Phil          http://www.gerbil.org.uk/
Please reply to rpg 'at' mrc-lmb.cam.ac.uk

'There's a fine line between genius and just farting around'




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