Re: [ccp4bb] Semet in non-auxotrophic strains

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

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Subject: Re: Semet in non-auxotrophic strains
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-11-03

Jacob,

We routinely express SeMet proteins in native E. coli without any trouble.
The method is a derivative of Doublie's. If you want, I would be more than
happy to send you a protocol (we've modified the process to gain biomass
yield).

Artem

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Jacob
Keller
Sent: Monday, November 03, 2008 12:56 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Semet in non-auxotrophic strains

Dear Crystallographers,

I have been using an E coli strain called Tuner (DE3) pLacI, which is not
auxotrophic for methionine, nor is there an auxotrophic analogue available.
It may be that my protein can be expressed in other strains, but this one
works well right now, and it is usually better to stay with what works well.

Has anybody tried growing semet protein in non-auxotrophic strains, or are
there tricks to using non-auxotrophs for semet protein?

Thanks,

Jacob

*******************************************
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
*******************************************

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

Previous message:
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