expression in E.coli

Warren Gallin wgallin at
Wed Mar 16 11:04:50 EST 1994

In Article <plyhmb.21.000D358E at>, plyhmb at
(Mark Bond) wrote:
>Does anyone have any suggestions as how to maximize eucayotic gene expression 
>in E.coli.  I have been trying to induce expression of a GST/talin fusion 
>protein and have only been able to produced small amounts of protein.  I know 
>that at 37oC many foreign protein in E.coli aggregate and this can be 
>partially overcome by incubating the cells at a lower temperature.  Is this 
>the only method that I can use to incease the protein yield.

   It ssounds from your post like you are viewing the aggregate forms as
lost.  Don't.  One of the standard ways of preparing the fusion proteins is
to isolate the inclusion bodies and extract them with a strong denaturant. 
Check Current Protocols in Molecular Biology for the details.
   If your problem is degradation, on the other hand, try using host
bacteria that have been engineered to remove endogenous proteases (if you
haven't already).

Warren Gallin,
Department of Zoology, University of Alberta
wgallin at

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