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'
