pEt-Expression-Mystery

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Mon Aug 3 08:31:49 EST 1998


Lars Komorowski <larskomo at physik.mu-luebeck.de> wrote:
> I am trying to overexpress a 17kDa-Protein in E.coli BL21 with Novagens
> pET24a Vector. I inoculate my LB medium from an overnight culture, let the
> culture grow until the OD reaches 0.6 and induce with 1 mM IPTG. After one
> night there is a band at 25 kDa in SDS-PAGE made with total cell protein.

If your protein is unstable, it will not survive overnight induction.
Can you detect your protein by western blotting?

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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