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Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overla...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Sociedade Brasileira de Genética
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168189/ https://www.ncbi.nlm.nih.gov/pubmed/21931521 http://dx.doi.org/10.1590/S1415-47572011005000022 |
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author | Teh, Ser Huy Fong, Mun Yik Mohamed, Zulqarnain |
author_facet | Teh, Ser Huy Fong, Mun Yik Mohamed, Zulqarnain |
author_sort | Teh, Ser Huy |
collection | PubMed |
description | The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo. |
format | Online Article Text |
id | pubmed-3168189 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Sociedade Brasileira de Genética |
record_format | MEDLINE/PubMed |
spelling | pubmed-31681892011-09-19 Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris Teh, Ser Huy Fong, Mun Yik Mohamed, Zulqarnain Genet Mol Biol Genetics of Microorganisms The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo. Sociedade Brasileira de Genética 2011-07-01 2011 /pmc/articles/PMC3168189/ /pubmed/21931521 http://dx.doi.org/10.1590/S1415-47572011005000022 Text en Copyright © 2011, Sociedade Brasileira de Genética. Printed in Brazil License information: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Genetics of Microorganisms Teh, Ser Huy Fong, Mun Yik Mohamed, Zulqarnain Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title | Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title_full | Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title_fullStr | Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title_full_unstemmed | Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title_short | Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris |
title_sort | expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in pichia pastoris |
topic | Genetics of Microorganisms |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168189/ https://www.ncbi.nlm.nih.gov/pubmed/21931521 http://dx.doi.org/10.1590/S1415-47572011005000022 |
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