( This in turn leads to slowing or abort...)New BL21-CodonPlus Cells Correct Codon Bias in GC-Rich Genomes

This in turn, leads to slowing or abortion of the translation process and subsequent degradation of the mRNA.⁴ When the 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 gene).

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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