This in turn, leads to slowing or abortion of the
translation process and subsequent degradation of the mRNA.4
problem of codon bias causes frameshifts, codon skipping, or misincorporations,
it may escape detection. More commonly, manifestations of codon bias include no
or low protein expression and the presence of aborted translation products.
Previously, the typical remedies for overcoming codon bias were to alter the
codon specifications of a target gene by site-directed mutagenesis or to
circumvent the problem by switching to a eukaryotic expression system.
To make it easier to resolve the codon bias problem, Stratagene released
BL21-CodonPlus-RIL competent cells,5 which are derivatives of the
BL21-Gold series. BL21-CodonPlus-RIL cells contain a ColE1-compatible vector
with extra copies of the argU, ileY, and leuW tRNA genes.
These tRNAs recognize arginine, isoleucine, and leucine codons (AGA/AGG, AUA,
and CUA), respectively. Host cell expression of these tRNAs can lead to
increased synthesis of recombinant proteins, as translation is no longer limited
by the availability of tRNAs that recognize rare codons. However, using these
codons is predominantly a problem in organisms with AT-rich genomes (Table 1).
For organisms whose genomes are GC rich, the problematic codons for expression
are the arginine codon AGG (recognized by the tRNA product of the argU
gene) and the proline codon CCC (recognized by the tRNA product of the proL
Hence, Stratagene designed BL21-CodonPlus-RP and BL21-CodonPlus(DE3)-RP
competent cells to solve this dilemma. These cells include a ColE1-compatible
expression plasmid that contains the tRNA genes argU and proL. The
corresponding tRNAs recognize the arginine codons, AGA/AGG, and the proline
codon, CCC, respectively.
Rare Codons and Protein Production
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