Recombinant protein expression in Escherichia coli: advances and challenges

doi:10.3389/fmicb.2014.00172

Review

. 2014 Apr 17:5:172.

doi: 10.3389/fmicb.2014.00172. eCollection 2014.

Recombinant protein expression in Escherichia coli: advances and challenges

Germán L Rosano¹, Eduardo A Ceccarelli¹

Affiliations

Affiliation

¹ Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas Rosario, Argentina ; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario Rosario, Argentina.

PMID: 24860555
PMCID: PMC4029002
DOI: 10.3389/fmicb.2014.00172

Review

Recombinant protein expression in Escherichia coli: advances and challenges

Germán L Rosano et al. Front Microbiol. 2014.

. 2014 Apr 17:5:172.

doi: 10.3389/fmicb.2014.00172. eCollection 2014.

Authors

Germán L Rosano¹, Eduardo A Ceccarelli¹

Affiliation

¹ Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas Rosario, Argentina ; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario Rosario, Argentina.

PMID: 24860555
PMCID: PMC4029002
DOI: 10.3389/fmicb.2014.00172

Abstract

Escherichia coli is one of the organisms of choice for the production of recombinant proteins. Its use as a cell factory is well-established and it has become the most popular expression platform. For this reason, there are many molecular tools and protocols at hand for the high-level production of heterologous proteins, such as a vast catalog of expression plasmids, a great number of engineered strains and many cultivation strategies. We review the different approaches for the synthesis of recombinant proteins in E. coli and discuss recent progress in this ever-growing field.

Keywords: E. coli expression strains; Escherichia coli; affinity tags; expression plasmid; inclusion bodies; recombinant protein expression.

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Figures

**FIGURE 1**
**Anatomy of an expression vector. The figure depicts the major features present in common expression vectors**. All of them are described in the text. The affinity tags and coding sequences for their removal were positioned arbitrarily at the N-terminus for simplicity. MCS, multiple cloning site. Striped patterned box: coding sequence for the desired protein.

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Comment in

Commentary: A tribute to Dr. Paul Berg (1926-2023) American biochemist, Nobel Laureate and discoverer of recombinant DNA technology, vaccine and genetic engineering.
Lukiw WJ. Lukiw WJ. Front Cell Dev Biol. 2023 May 18;11:1210530. doi: 10.3389/fcell.2023.1210530. eCollection 2023. Front Cell Dev Biol. 2023. PMID: 37274735 Free PMC article. No abstract available.

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References

1. Adrio J. L., Demain A. L. (2010). Recombinant organisms for production of industrial products. Bioeng. Bugs 1 116–131 10.4161/bbug.1.2.10484 - DOI - PMC - PubMed
1. Alibolandi M., Mirzahoseini H. (2011). Chemical assistance in refolding of bacterial inclusion bodies. Biochem. Res. Int. 2011:631607 10.1155/2011/631607 - DOI - PMC - PubMed
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1. Angov E., Legler P. M., Mease R. M. (2011). Adjustment of codon usage frequencies by codon harmonization improves protein expression and folding. Methods Mol. Biol. 705 1–13 10.1007/978-1-61737-967-3_1 - DOI - PubMed
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[1] Adrio J. L., Demain A. L. (2010). Recombinant organisms for production of industrial products. Bioeng. Bugs 1 116–131 10.4161/bbug.1.2.10484 - DOI - PMC - PubMed

[2] Adrio J. L., Demain A. L. (2010). Recombinant organisms for production of industrial products. Bioeng. Bugs 1 116–131 10.4161/bbug.1.2.10484 - DOI - PMC - PubMed

[3] Alibolandi M., Mirzahoseini H. (2011). Chemical assistance in refolding of bacterial inclusion bodies. Biochem. Res. Int. 2011:631607 10.1155/2011/631607 - DOI - PMC - PubMed

[4] Alibolandi M., Mirzahoseini H. (2011). Chemical assistance in refolding of bacterial inclusion bodies. Biochem. Res. Int. 2011:631607 10.1155/2011/631607 - DOI - PMC - PubMed

[5] Andreev Y. A., Kozlov S. A., Vassilevski A. A., Grishin E. V. (2010). Cyanogen bromide cleavage of proteins in salt and buffer solutions. Anal. Biochem. 407 144–146 10.1016/j.ab.2010.07.023 - DOI - PubMed

[6] Andreev Y. A., Kozlov S. A., Vassilevski A. A., Grishin E. V. (2010). Cyanogen bromide cleavage of proteins in salt and buffer solutions. Anal. Biochem. 407 144–146 10.1016/j.ab.2010.07.023 - DOI - PubMed

[7] Angov E., Legler P. M., Mease R. M. (2011). Adjustment of codon usage frequencies by codon harmonization improves protein expression and folding. Methods Mol. Biol. 705 1–13 10.1007/978-1-61737-967-3_1 - DOI - PubMed

[8] Angov E., Legler P. M., Mease R. M. (2011). Adjustment of codon usage frequencies by codon harmonization improves protein expression and folding. Methods Mol. Biol. 705 1–13 10.1007/978-1-61737-967-3_1 - DOI - PubMed

[9] Atlas R. M. (2004). Handbook of Microbiological Media, 3rd Edn. Boca Raton, FL: Taylor & Francis

[10] Atlas R. M. (2004). Handbook of Microbiological Media, 3rd Edn. Boca Raton, FL: Taylor & Francis

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Recombinant protein expression in Escherichia coli: advances and challenges

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Recombinant protein expression in Escherichia coli: advances and challenges

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