[ccp4bb] Bacterial induction at 18C

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From: Kay Diederichs Kay {- dot -} Diederichs {- at -} UNI-KONSTANZ {- dot -} DE
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Subject: Re: Bacterial induction at 18C
From: Christopher Law chrisjlaw {- at -} GMAIL {- dot -} COM
Date: 2008-04-30

Subject: Bacterial induction at 18C
From: Raji Edayathumangalam redayath {- at -} MAIL {- dot -} ROCKEFELLER {- dot -} EDU
Date: 2008-04-30

Hi Folks,

I am working with E. coli cells co-transformed with two plasmids and I find that my cells lyse
following overnight inductions at 18C. I suspect (among many things) that Ampicillin+
Chloramphenicol+ Kanamycin in the medium may be the source of my woes.

My colleagues have suggested growing cultures at 18C, say for 4-6h instead. Has anyone had
reasonable protein expression levels by inducing cultures at 18C for 6h? From what I understand, the
E. coli doubling time is manyfold longer than at 37C. But I thought I'd ask.

I am already playing with lowering and/or doing away with the antibiotics.

Any suggestions wrt 18C? The protein is insoluble at 30C.

Thanks.
Raji

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CCP4bb <-- 2008 <-- April 2008 <-- 30 April 2008

Previous message:
Subject: Re: xds installation
From: Kay Diederichs Kay {- dot -} Diederichs {- at -} UNI-KONSTANZ {- dot -} DE
Date: 2008-04-30

Next message:
Subject: Re: Bacterial induction at 18C
From: Christopher Law chrisjlaw {- at -} GMAIL {- dot -} COM
Date: 2008-04-30