Have a look at this paper:
http://www.ncbi.nlm.nih.gov/pubmed?term=18289875
the bottom line appears to be that, in most cases, co-transforming with
plasmids expressing rare tRNAs is just as good as codon optimizing the gene for
your protein of interest. In addition to the financial aspects already touched
upon by Tassos, we even had cases where codon optimizing a gene actually
completely abolished its expression, presumably by causing havoc at the mRNA
level. This obviously won't happen if you co-express rare codon tRNAs...
HTH, Luca
----------------------------------------------------------------
Luca Jovine, Ph.D.
Assistant Professor & EMBO Young Investigator
Karolinska Institutet
Department of Biosciences and Nutrition & Center for Biosciences
Hälsovägen 7, SE-141 57 Huddinge, Sweden
Voice: +46.(0)8.524-81136 FAX: +46.(0)8.6081-501
E-mail: luca.jov...@ki.se
W3: http://jovinelab.org
----------------------------------------------------------------
On Sep 30, 2011, at 19:13 , Segelke, Brent W. wrote:
> 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:abba...@gmail.com] On Behalf Of Anastassis
> Perrakis
> Sent: Friday, September 30, 2011 10:08 AM
> To: Segelke, Brent W.
> Cc: CCP4BB@JISCMAIL.AC.UK
> 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." <segel...@llnl.gov> 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:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Tim Keys
> Sent: Friday, September 30, 2011 8:29 AM
> To: CCP4BB@JISCMAIL.AC.UK
> 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: CCP4BB@JISCMAIL.AC.UK
> 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
> pat.l...@drexelmed.edu
>
>
>
