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Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β
A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stabl...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Published by Elsevier Ltd.
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128424/ https://www.ncbi.nlm.nih.gov/pubmed/21459017 http://dx.doi.org/10.1016/j.cyto.2010.12.002 |
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author | Chen, Weiye Cao, Wenyan Zhao, Huijun Hu, Qianqian Qu, Linmao Hu, Sen Ge, Jinying Wen, Zhiyuan Wang, Xijun Li, HaoBo Huang, Kehe Bu, Zhigao |
author_facet | Chen, Weiye Cao, Wenyan Zhao, Huijun Hu, Qianqian Qu, Linmao Hu, Sen Ge, Jinying Wen, Zhiyuan Wang, Xijun Li, HaoBo Huang, Kehe Bu, Zhigao |
author_sort | Chen, Weiye |
collection | PubMed |
description | A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-β could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-β has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-β fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-β. |
format | Online Article Text |
id | pubmed-7128424 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Published by Elsevier Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71284242020-04-08 Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β Chen, Weiye Cao, Wenyan Zhao, Huijun Hu, Qianqian Qu, Linmao Hu, Sen Ge, Jinying Wen, Zhiyuan Wang, Xijun Li, HaoBo Huang, Kehe Bu, Zhigao Cytokine Article A CHO cell clone (CHO-PoIFN-β) with stable porcine IFN-β expression under control of CMV promoter was selected under G418 pressure. In a 25 cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-β reached 2.3 × 10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-β could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-β has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-β fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-β. Published by Elsevier Ltd. 2011-06 2011-04-01 /pmc/articles/PMC7128424/ /pubmed/21459017 http://dx.doi.org/10.1016/j.cyto.2010.12.002 Text en Copyright © 2010 Published by Elsevier Ltd. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Chen, Weiye Cao, Wenyan Zhao, Huijun Hu, Qianqian Qu, Linmao Hu, Sen Ge, Jinying Wen, Zhiyuan Wang, Xijun Li, HaoBo Huang, Kehe Bu, Zhigao Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title | Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title_full | Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title_fullStr | Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title_full_unstemmed | Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title_short | Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-β |
title_sort | establishment of a stable cho cell line with high level expression of recombinant porcine ifn-β |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128424/ https://www.ncbi.nlm.nih.gov/pubmed/21459017 http://dx.doi.org/10.1016/j.cyto.2010.12.002 |
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