Is there a general consensus that this is true? I’ve heard exactly the opposite, i.e., that codon optimization rarely gives you dramatically improved yields of soluble protein. Are there any published studies on this topic? This seems like something that might come out of one of the SG centers.
Brent From: Anastassis Perrakis [mailto:[email protected]] On Behalf Of Anastassis Perrakis Sent: Friday, September 30, 2011 10:08 AM To: Segelke, Brent W. Cc: [email protected] Subject: Re: [ccp4bb] is codon optimization worth it? Well, codon optimization is not really trouble, it's money. The money are worth it usually anyway, since the optimized genes are easy to clone if you make many constructs out of one gene, as you better do anyway ... Compared with downstream expenses, optimized genes are these days almost always worth the trouble... A. Sent from my iPad On 30 Sep 2011, at 19:02, "Segelke, Brent W." <[email protected]<mailto:[email protected]>> wrote: To me, the key question would seem to be, if I can’t win them all, how many more do I win if I go to the trouble? Brent From: CCP4 bulletin board [mailto:[email protected]] On Behalf Of Tim Keys Sent: Friday, September 30, 2011 8:29 AM To: [email protected]<mailto:[email protected]> Subject: Re: [ccp4bb] is codon optimization worth it? We codon optimised a poorly expressed gene from neisseria meningitides based on a codon usage table derived from the Welch (etal, 2009) paper below. The optimisation is specifically for overexpression in BL21 (DE3). The optimised gene increased protein expression by at least a factor of 10, and changed (somewhat reduced) the degradation pattern we observed. Unfortunately it didn't do anything to improve the folding (ie. we ended up with lots of half-folded, semi-soluble protein). With other neisserial derived proteins we have had an almost undetectable effect. You can't win 'em all. Cheers, Tim Design Parameters to Control Synthetic Gene Expression in Escherichia coli Welch et al, PlosONE 2009 Medizinische Hochschule Hannover Zelluläre Chemie, OE 4330 Zentrum Biochemie Carl-Neubergstr. 1 30625 Hannover -----Original Message----- From: CCP4 bulletin board on behalf of Patrick Loll Sent: Fri 30.09.2011 16:49 To: [email protected]<mailto:[email protected]> Subject: [ccp4bb] is codon optimization worth it? 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) Thanks, Pat --------------------------------------------------------------------------------------- Patrick J. Loll, Ph. D. Professor of Biochemistry & Molecular Biology Director, Biochemistry Graduate Program Drexel University College of Medicine Room 10-102 New College Building 245 N. 15th St., Mailstop 497 Philadelphia, PA 19102-1192 USA (215) 762-7706 [email protected]<mailto:[email protected]>
