Re: [ccp4bb] Semet in non-auxotrophic strains (summary)

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

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

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From: nurizzo nurizzo {- at -} ESRF {- dot -} FR
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Subject: Re: Semet in non-auxotrophic strains (summary)
From: Vangelis Christodoulou e {- dot -} christodoulou {- at -} NKI {- dot -} NL
Date: 2008-11-04

While staying on the subject, I found the following paper very useful
when trying to produce Se-Met labeled protein in HEK293 Flp-In.

Barton et al. Highly efficient selenomethionine labeling of
recombinant proteins produced in mammalian cells. Protein Sci (2006)
vol. 15 (8) pp. 2008-13

Vangelis

--
Vangelis Christodoulou
Analist C
Anastassis (Tassos) Perrakis lab
The Netherlands Cancer Institute (NKI) B8.038
Plesmanlaan 121, 1066 CX, Amsterdam
The Netherlands

tel: +31 (0)20 512 1963
fax: +31 (0)20 512 1954
Website: xtal.nki.nl/Tassos_group/

On 4 Nov 2008, at 03:27, Artem Evdokimov wrote:

> http://www.xtals.org/pdfs/selenium.pdf
>
> (sorry, for some reason first email went out w/o the hyperlink)
>
> Artem
>
> -----Original Message-----
> From: Artem Evdokimov [mailto:artem@xtals.org]
> Sent: Monday, November 03, 2008 9:26 PM
> To: 'Jacob Keller'; 'CCP4BB@JISCMAIL.AC.UK'
> Subject: RE: [ccp4bb] Semet in non-auxotrophic strains (summary)
>
> By popular request I've placed our modified Se-Met method online:
>
>
> -----Original Message-----
> From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf
> Of Jacob
> Keller
> Sent: Monday, November 03, 2008 8:13 PM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: Re: [ccp4bb] Semet in non-auxotrophic strains (summary)
>
> Hello all,
>
> Prompted by Kay Diederichs, herein follows a summary of the responses
> gleaned from the CCP4BB. The wiki has also been edited to include the
> summary:
>
> http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_Se
> Met_labeled_proteins
>
> Summary:
>
> 1. Protocols for both auxotrophs and non-auxotrophs:
> Doublié, S. (1997). Methods Enzymol. 276, 523-532
>
> 2. Two protocols for semet labelling:
> http://www.doe-mbi.ucla.edu/local/protocols
>
> 3. Another non-auxotroph protocol. G. Fritz says this recipe could
> probably
> be made in one vessel for immediate use, except for the aromatic
> aa's which
> might dissolve better at basic pH:
> http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_Se
> Met_labeled_proteins
>
> 4. Original literature source for met synthesis silencing (for use
> with
> non-auxotrophs):
> Van Duyne et al. J. Mol. Biol. 229, 105-124 (1993).
>
> 5. Autoinduction semet protocol (allegedly complicated recipe):
> Studier FW (2005) Protein production by auto-induction in high density
> shaking cultures. Protein Expr. Purif. 41, 207-234
>
> 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: Semet in non-auxotrophic strains (summary)
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-11-03

Next message:
Subject: ESRF School - 2nd - 5th February 2009
From: nurizzo nurizzo {- at -} ESRF {- dot -} FR
Date: 2008-11-04