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:[EMAIL PROTECTED] 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:[EMAIL PROTECTED] 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: [EMAIL PROTECTED] *******************************************
