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Re: [ccp4bb] Bacterial induction at 18C
 

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CCP4bb <-- 2008 <-- April 2008 <-- 30 April 2008
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Subject: Bacterial induction at 18C
From: Raji Edayathumangalam redayath {- at -} MAIL {- dot -} ROCKEFELLER {- dot -} EDU
Date: 2008-04-30 		Next message:
Subject: Re: Bacterial induction at 18C
From: Guenter Fritz guenter {- dot -} fritz {- at -} UNI-KONSTANZ {- dot -} DE
Date: 2008-04-30


Subject: Re: Bacterial induction at 18C
From: Christopher Law chrisjlaw {- at -} GMAIL {- dot -} COM
Date: 2008-04-30
Are you sure it is not a phage infection that is causing lysis of your cells?

Christopher J. Law, PhD
Skirball Institute of Biomolecular Medicine,
Structural Biology (3-5),
New York University Medical Center,
540 First Avenue,
New York, NY 10016, USA.

2008/4/30 Raji Edayathumangalam :
> 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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