Rare arginine codon

CARLOS DUARTE Carlos.Duarte at CIGB.EDU.CU
Sat Sep 12 13:21:22 EST 1998


Tomer
Plasmids encoding for dnaY that is tRNA for codon AGA has been 
used to improve expresion of genes with high number of AGA and AGG 
codons.
You can consult the following references
Annals New York Acad Sci, 1996, 782:79-86
Cell , 1985, 45:453-459
Gene 1989, 85:109-114
Biotechniques, 1995, 19,196-200

One of such plasmids should be in my institute but I am 
sure that it will be easier for you to get it from some other lab in the 
US
Hope that helps


Bryan L. Ford wrote:
> This approach seems difficult since the imbalance of tRNA will likewise
> influence translation efficiency of the native E. coli genes. Let me ask
> if
> there is any reason that you cannot modify the rare codon into a
> preferred E. coli codon, using the quite straightforward (that is
> *easy*) methods of site-specific or site-directed mutagenesis?  For
> example (using data for E. coli taken from the Codon Usage Database
> found online at http://www.dna.affrc.go.jp/~nakamura/codon.html ) we see
> that AGG is the rarest at 1.5 per thousand, and CGA is relatively rare
> at 3.6/k. Both can be changed into frequently used codons with single
> base changes, that is to AGC with a usage of 15.7/k and to CGC, which
> happens to have the highest usage of all six arginyl codons at 21.5/k.
> 
> Let me know if you need more information about about site-specific
> mutagenesis protocols.
> 
> -Bryan


Bryan,

I did not mention that this rare arginine codon is quite frequent in our
gene (more than 20 occurances).  I'm affraid that it might not be either
cost or time effective to mutate the codon within the gene.

Thanks for your input.

Sincerely,
Tomer

Carlos A. Duarte
Centro de Ingenieria Genetica y Biotecnologia 
Division de Vacunas
Apdo 6162, Ciudad Habana CP10600
Cuba
Tel 53-7-218008
Fax 53-7-218070-336008



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