Abstract
Over the past two decades, prokaryotic expression systems have been widely exploited for the bioproduction of many therapeutic proteins. Much of the success can be attributed to the implementation of basic principles of prokaryotic protein translation and protein folding to the problems of heterologous expression (e.g. codon usage substitutions, tRNA isoacceptor co-expression, chaperone co-expression); however, expression in a heterologous host still remains an empirical process. To improve heterologous protein expression further we have developed an algorithm termed “codon harmonization” that best approximates codon usage frequencies from the native host and adjusts these for use in the heterologous system. The success of this methodology may be due to improved protein folding during translation. Although so far exclusively applied to Escherichia coli, codon harmonization may provide a general strategy for improving the expression of soluble, functional proteins during heterologous host expression.
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Acknowledgements
This work was supported by the United States Agency for International Development, Project Number 936–6001, Award Number AAG-P-00–98–00006, Award Number AAG-P-00-98-00005 and by the United States Army Medical Research and Materiel Command. The authors acknowledge the conceptual contributions of Drs. Jeffrey A. Lyon and Randall L. Kincaid for “codon harmonization”. From the Division of Biochemistry, we thank Ms. Amy Michels for editorial assistance.
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Angov, E., Legler, P.M., Mease, R.M. (2011). Adjustment of Codon Usage Frequencies by Codon Harmonization Improves Protein Expression and Folding. In: Evans, Jr., T., Xu, MQ. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 705. Humana Press. https://doi.org/10.1007/978-1-61737-967-3_1
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DOI: https://doi.org/10.1007/978-1-61737-967-3_1
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