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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:[log in to unmask]] On Behalf Of Jacob Keller Sent: Monday, November 03, 2008 12:56 PM To: [log in to unmask] 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: [log in to unmask] *******************************************