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]> 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] > 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] > 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] >
