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Codon Bias-Adjusted BL21 Derivatives for Protein Expression


High-level expression of difficult heterologous proteins

Carsten-Peter Carstens Anna Waesche
Stratagene

Stratagene has created variants of the Epicurian Coli BL21-Gold cells * called the BL21-CodonPlus -RIL series** of strains, which contain extra copies of the E. coli argU, ileY, and leuW tRNA genes. This modification allows for a high-level expression of proteins that are difficult to express in conventional E. coli hosts due to the codon bias of the gene of interest. The BL21-CodonPlus-RIL strains can eliminate the need to move the gene of interest into eukaryotic expression systems.

BL21-Gold(DE3) cells are the strain of choice for high-level production of heterologous proteins expressed from the T7-promoter. These strains have been engineered to contain an inducible T7-RNA polymerase. They are also EndA- to allow high-quality miniprep DNA preparation.1 BL21 competent cells are derivatives of E. coliB and lack the Lon protease and the ompT protease genes that may cause instability and degradation of the recombinant protein.

Table 1
Rarest Codons (from E. coli) Preferred by Selected Organisms

Codon frequencies are expressed as codons used per 1000 codons encountered. A complete compilation of codon usage of the sequences placed in the gene bank database can be found at http://www.dna.affrc.go.jp/~nakamura/codon.html Codon frequencies of more than 15 codons/1000 codons are shown in bold to help identify a codon bias that may cause problems for high-level expression in E. coli. The arginine codons AGG and AGA are recognized by the same tRNA (pr
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