age--
Date: 30-apr-2008 12:30:50 -0400
From: "Guenter Fritz" <[EMAIL PROTECTED]>
To: <[EMAIL PROTECTED]>
Cc:
Subject: Re: [ccp4bb] Bacterial induction at 18C
Raji,
I am working with E. coli cells co-transformed with two plasmids and I find
that my cells
Raji
aside from the possibilites of toxic protein as already mentioned..
we had great results with overnight induction at 20oC for a protein that was
somewhat insoluble at 37oC. One thing that might work for you is to grow the
cells to OD 0.5 then lower the temperature to say 18 or 20. After
ncluded Message--
>Date: 30-apr-2008 12:30:50 -0400
>From: "Guenter Fritz" <[EMAIL PROTECTED]>
>To: <[EMAIL PROTECTED]>
>Cc:
>Subject: Re: [ccp4bb] Bacterial induction at 18C
>
>Raji,
>
>>
>> I am working with E. coli cells co-tra
-8001
[EMAIL PROTECTED]
www.accelagen.com
-Original Message-
From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Raji
Edayathumangalam
Sent: Wednesday, April 30, 2008 8:40 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Bacterial induction at 18C
Hi Folks,
I am working with E.
Dear Raji,
I have expressed proteins in E. coli at 18ºC (and below that) with
Amp+Kan+Chl and I've never had any problem with cell lysis. I usually grow
cells at 37ºC and then cool the cultures down to the desired temperature
before induction. I don't recomend growing your cultures at low
temperat
Raji,
I am working with E. coli cells co-transformed with two plasmids and I find
that my cells lyse
following overnight inductions at 18C.
Sounds more like a phage contamination. The phage becomes active as soon
as the cells "energy level" decreases, e.g upon induction. We had once
the sam
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 <[EMAIL PROTE
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 grow
