Rosetta strains carry a plasmid that supplies several tRNAs that match codons that are rare in E. coli. This is quite different than being explicitly optimized to express mammalian proteins. We (the Center for Eukaryotic Structural Genomics, a PSI-1 and PSI-2 center) used strains with the same tRNA plasmid (either pRARE or pRARE2) to express proteins from yeast, Arabidopsis, some thermophilic eukaryotes, mouse, frog, human and zebrafish proteins. It seemed to work just fine for all of them. On Sep 30, 2011, at 9:55 AM, Ed Pozharski wrote:
> On Fri, 2011-09-30 at 10:49 -0400, Patrick Loll wrote: >> Has anyone encountered a case in which a construct with the native >> sequence expressed poorly (or not at all?) in Rosetta(DE3), but the >> corresponding construct with a codon-optimized sequence expressed >> well? (The gene in question is from cerevesiae) >> > > Wait, isn't Rosetta optimized for mammalian genes, not yeast (but maybe > codon bias in yeast matches that)? > > A sideways suggestion would be to express in yeast. Extra bonus - it > smells like beer :) > > -- > "Hurry up before we all come back to our senses!" > Julian, King of Lemurs
