Help on Prokayrotic Protein Expression

[Per Mygind]

AKi Hoji wrote:

> Hi All,
>
> I cloned the cDNA encoding 40kd protein in pET23a, and have tried to
> produce this protein in BL21(DE3) upon IPTG induction, which should
> accumulate in inclusion bodies (for refolding purpose later).  However, I
> have been getting very little expression (compared to my positive control)
> even for 3-4 hrs induction at 0.4mM IPTG.  This protein is  reported to be
> non-toxic to bacteria, and in fact they appear to look fine after
> induction.  I appreciate any suggestions at this point.  Thanks in
> advance.
> --
> Aki Hoji (akhst7+ at pitt.edu)
> Dept. Infectious Diseases & Microbiology
> University of Pittsburgh

Try another strain, e.g. Bl21-DE3-pLysS

Regards

Per Mygind

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If you are are not part of the solution, you are part of the precipitate
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Per Mygind, Cand.scient

Department of Medical Microbiology and Immunology
The Bartholin Building, University of Aarhus, Denmark
phone : 89 42 17 47, fax   : 86 19 61 28

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It's hard work and great art to make life not so serious
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