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_SeMet_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_SeMet_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
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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]
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