Re: [ccp4bb] Shredded E coli pellets

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Shredded E coli pellets
From: Paul Leonard leonard {- at -} CABM {- dot -} RUTGERS {- dot -} EDU
Date: 2009-07-02

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Subject: Re: Shredded E coli pellets
From: "Van Den Berg, Bert" Lambertus {- dot -} VandenBerg {- at -} UMASSMED {- dot -} EDU
Date: 2009-07-02

Subject: Re: Shredded E coli pellets
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2009-07-02

Yes,

This is 95% likely to be the dreaded T1 phage (well, OK - any lytic phage
that infects your culture, really). The almost-only other alternative is
that your protein lyses the cells.

Dealing with phage infection can be tough especially in the multi-user
environment and doubly so if you make your own competent cells...

Artem

"Nothing is built on stone; all is built on sand, but we must build as if
the sand were stone"
Jorge Luis Borges

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Jacob
Keller
Sent: Thursday, July 02, 2009 11:42 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Shredded E coli pellets

Dear crystallographers,

I recently expressed some new constructs, and found after my usual
expression protocol that the cell pellets were not compacted at the bottom
corner of the bottles us usual, but were instead smeared as a film on the
side, and further, were somewhat clumpy, like clots, and with a smaller
pellet in the usual location. The centrifugation was exactly as usual. I
noticed that there was also a bit more foam in the medium than usual, but I
am not convinced that this was the issue, although it might be a symptom. My

suspicion is that the constructs are lethal and cause cell lysis, but I am
not sure. Has anybody seen this phenomenon before, and gotten to the bottom
of it?

Jacob Keller

*******************************************
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: j-keller2@northwestern.edu
*******************************************

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: Shredded E coli pellets
From: Paul Leonard leonard {- at -} CABM {- dot -} RUTGERS {- dot -} EDU
Date: 2009-07-02

Next message:
Subject: Re: Shredded E coli pellets
From: "Van Den Berg, Bert" Lambertus {- dot -} VandenBerg {- at -} UMASSMED {- dot -} EDU
Date: 2009-07-02