You can express Se-Met protein in non-auxotrophs using metabolic
inhibition (Doublie. Methods Enzymol. 276, 523-530, 1997 and Van Duyne
et al. J. Mol. Biol. 229, 105-124, 1993).
The Studier auto-induction paper (Studier, Prot. Exp. Purif. 41,
207-234, 2005) describes an auto-induction Se-Met medium that gives
sufficient incorporation of Se-Met for phasing in non-auxotrophs.
Good luck,
Cynthia
On Nov 3, 2008, at 12:56 PM, Jacob Keller wrote:
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: [EMAIL PROTECTED]
*******************************************
____________________
Cynthia Kinsland, Ph.D.
Cornell University
Protein Facility Director
607-255-8844
