Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system

Current research focuses on the soluble and high-level expression of biologically active recombinant human IL-29 protein in Escherichia coli. The codon-optimized IL-29 gene was cloned into the Champion™ pET SUMO expression system downstream of the SUMO tag under the influence of the T7 lac promoter....

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Detalles Bibliográficos
Autores principales: Munir, Ayesha, Ahmed, Nadeem, Akram, Muhammad, Fujimura, Nao Akusa, Tahir, Saad, Malik, Kausar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2023
Materias:
Original Research Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10235849/
https://www.ncbi.nlm.nih.gov/pubmed/37266881
http://dx.doi.org/10.1007/s10529-023-03402-x
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author Munir, Ayesha
Ahmed, Nadeem
Akram, Muhammad
Fujimura, Nao Akusa
Tahir, Saad
Malik, Kausar
author_facet Munir, Ayesha
Ahmed, Nadeem
Akram, Muhammad
Fujimura, Nao Akusa
Tahir, Saad
Malik, Kausar
author_sort Munir, Ayesha
collection PubMed
description Current research focuses on the soluble and high-level expression of biologically active recombinant human IL-29 protein in Escherichia coli. The codon-optimized IL-29 gene was cloned into the Champion™ pET SUMO expression system downstream of the SUMO tag under the influence of the T7 lac promoter. The expression of SUMO-fused IL-29 protein was compared in E. coli Rosetta 2(DE3), Rosetta 2(DE3) pLysS, and Rosetta-gami 2(DE3). The release of the SUMO fusion partner resulted in approximately 98 mg of native rhIL-29 protein with a purity of 99% from 1 l of fermentation culture. Purified rhIL-29 was found to be biologically active, as evaluated by its anti-proliferation assay. It was found that Champion™ pET SUMO expression system can be used to obtained high yield of biologically active soluble recombinant human protein compared to other expression vector.
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spelling pubmed-102358492023-06-06 Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system Munir, Ayesha Ahmed, Nadeem Akram, Muhammad Fujimura, Nao Akusa Tahir, Saad Malik, Kausar Biotechnol Lett Original Research Paper Current research focuses on the soluble and high-level expression of biologically active recombinant human IL-29 protein in Escherichia coli. The codon-optimized IL-29 gene was cloned into the Champion™ pET SUMO expression system downstream of the SUMO tag under the influence of the T7 lac promoter. The expression of SUMO-fused IL-29 protein was compared in E. coli Rosetta 2(DE3), Rosetta 2(DE3) pLysS, and Rosetta-gami 2(DE3). The release of the SUMO fusion partner resulted in approximately 98 mg of native rhIL-29 protein with a purity of 99% from 1 l of fermentation culture. Purified rhIL-29 was found to be biologically active, as evaluated by its anti-proliferation assay. It was found that Champion™ pET SUMO expression system can be used to obtained high yield of biologically active soluble recombinant human protein compared to other expression vector. Springer Netherlands 2023-06-02 /pmc/articles/PMC10235849/ /pubmed/37266881 http://dx.doi.org/10.1007/s10529-023-03402-x Text en © The Author(s), under exclusive licence to Springer Nature B.V. 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Research Paper
Munir, Ayesha
Ahmed, Nadeem
Akram, Muhammad
Fujimura, Nao Akusa
Tahir, Saad
Malik, Kausar
Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title_full Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title_fullStr Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title_full_unstemmed Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title_short Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system
title_sort enhanced soluble expression of active recombinant human interleukin-29 using champion pet sumo system
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10235849/
https://www.ncbi.nlm.nih.gov/pubmed/37266881
http://dx.doi.org/10.1007/s10529-023-03402-x
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