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Volume 155, Issue 11

Overcoming codon-usage bias in heterologous protein expression in Free

Abstract

One of the limitations facing the development of into a successful vaccine vector is the inability of this bacterium to express high levels of heterologous proteins. In the present study, we have identified 12 codons deemed as rare codons in and seven other streptococcal species. tRNA genes encoding 10 of the 12 rare codons were cloned into a plasmid. The plasmid was transformed into strains of expressing the fusion protein SpaP/S1, the anti-complement receptor 1 (CR1) single-chain variable fragment (scFv) antibody, or the cyclophilin C18 protein. These three heterologous proteins contained high percentages of amino acids encoded by rare codons. The results showed that the production of SpaP/S1, anti-CR1 scFv and C18 increased by 2.7-, 120- and 10-fold, respectively, over the control strains. In contrast, the production of the streptococcal SpaP protein without the pertussis toxin S1 fragment was not affected by tRNA gene supplementation, indicating that the increased production of SpaP/S1 protein was due to the ability to overcome the limitation caused by rare codons required for the S1 fragment. The increase in anti-CR1 scFv production was also observed in following tRNA gene supplementation. Collectively, the findings in the present study demonstrate for the first time, to the best of our knowledge, that codon-usage bias exists in spp. and the limitation of heterologous protein expression caused by codon-usage bias can be overcome by tRNA supplementation.

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Keyword(s): CR1, complement receptor 1 , RT-PCR, reverse transcription PCR and scFv, single-chain variable fragment
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2009-11-01
2024-04-27
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Overcoming codon-usage bias in heterologous protein expression in Streptococcus gordonii
Microbiology 155, 3581 (2009); https://doi.org/10.1099/mic.0.030064-0
/content/journal/micro/10.1099/mic.0.030064-0
/content/journal/micro/10.1099/mic.0.030064-0
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