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Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts
BACKGROUND: Chloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplast transformation approach at high expression levels. However, high expression o...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9824920/ https://www.ncbi.nlm.nih.gov/pubmed/36611158 http://dx.doi.org/10.1186/s12896-022-00771-5 |
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author | Wang, Yunpeng Fan, Jieying Wei, Zhengyi Xing, Shaochen |
author_facet | Wang, Yunpeng Fan, Jieying Wei, Zhengyi Xing, Shaochen |
author_sort | Wang, Yunpeng |
collection | PubMed |
description | BACKGROUND: Chloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplast transformation approach at high expression levels. However, high expression of epidermal growth factor (EGF) in chloroplasts with the technology is still unavailable. RESULTS: The present work explored the high-level expression of recombinant EGF, a protein widely applied in many clinical therapies, in tobacco chloroplasts. In this work, homoplastic transgenic plants expressing fusion protein GFP-EGF, which was composed of GFP and EGF via a linker, were generated. The expression of GFP-EGF was confirmed by the combination of green fluorescent observation and Western blotting. The achieved accumulation of the recombinant fusion GFP-EGF was 10.21 ± 0.27% of total soluble proteins (1.57 ± 0.05 g kg(− 1) of fresh leaf). The chloroplast-derived GFP-EGF was capable of increasing the cell viability of the NSLC cell line A549 and enhancing the phosphorylation level of the EGF receptor in the A549 cells. CONCLUSION: The expression of recombinant EGF in tobacco chloroplasts via chloroplast transformation method was achieved at considerable accumulation level. The attempt gives a good example for the application of chloroplast transformation technology in recombinant pharmaceutical protein production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-022-00771-5. |
format | Online Article Text |
id | pubmed-9824920 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-98249202023-01-08 Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts Wang, Yunpeng Fan, Jieying Wei, Zhengyi Xing, Shaochen BMC Biotechnol Research BACKGROUND: Chloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplast transformation approach at high expression levels. However, high expression of epidermal growth factor (EGF) in chloroplasts with the technology is still unavailable. RESULTS: The present work explored the high-level expression of recombinant EGF, a protein widely applied in many clinical therapies, in tobacco chloroplasts. In this work, homoplastic transgenic plants expressing fusion protein GFP-EGF, which was composed of GFP and EGF via a linker, were generated. The expression of GFP-EGF was confirmed by the combination of green fluorescent observation and Western blotting. The achieved accumulation of the recombinant fusion GFP-EGF was 10.21 ± 0.27% of total soluble proteins (1.57 ± 0.05 g kg(− 1) of fresh leaf). The chloroplast-derived GFP-EGF was capable of increasing the cell viability of the NSLC cell line A549 and enhancing the phosphorylation level of the EGF receptor in the A549 cells. CONCLUSION: The expression of recombinant EGF in tobacco chloroplasts via chloroplast transformation method was achieved at considerable accumulation level. The attempt gives a good example for the application of chloroplast transformation technology in recombinant pharmaceutical protein production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-022-00771-5. BioMed Central 2023-01-07 /pmc/articles/PMC9824920/ /pubmed/36611158 http://dx.doi.org/10.1186/s12896-022-00771-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Wang, Yunpeng Fan, Jieying Wei, Zhengyi Xing, Shaochen Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title | Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title_full | Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title_fullStr | Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title_full_unstemmed | Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title_short | Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
title_sort | efficient expression of fusion human epidermal growth factor in tobacco chloroplasts |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9824920/ https://www.ncbi.nlm.nih.gov/pubmed/36611158 http://dx.doi.org/10.1186/s12896-022-00771-5 |
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