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problem with co-expression in e. coli


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#1 ad1980

ad1980

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Posted 26 January 2005 - 09:38 PM

Hi everybody.

I have a problem with co-expression in e. coli.

I co-transformed 2 vectors carrying 2 different proteins into e. coli
I used pRSET and pMON (p15A ori, kan resistant)

i tried 2 host strains: BL21(DE3) and BL21(DE3)pLysS
but...
protein in pRSET was expressed well in pLysS but one in pMON wasn't;
and protein in pMON was expressed well in BL21(DE3) but one in pRSET wasn't.

when co-expressed...
both proteins were not expressed well in both strains.
(couldn't see expression on SDS-PAGE)

how can i improve the expression (w/o changing vectors)?
would it help if i change hosts strains?
which strains can i use...?

I really appreciate if someone can help.

Edited by ad1980, 26 January 2005 - 09:44 PM.





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