Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts
Human papillomavirus (HPV) is the causal agent of cervical cancer, one of the most common causes of death in women worldwide, and its E7 antigen is the major candidate for a therapeutic vaccine. The large scale production of E7 by molecular farming that would lead to the development of a safe and in...
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paper:paper_10736085_v56_n11_p1021_Morgenfeld2023-06-08T16:04:58Z Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts Morgenfeld, Mauro Miguel Lentz, Ezequiel Matías Segretin, María Eugenia E7 antigen Fusion protein Human papillomavirus Molecular farming Thylakoid translocation Transplastomic tobacco Antigens Oncogenic viruses Plants (botany) Tobacco Vaccines Fusion proteins Human papillomavirus Large scale productions Molecular farming Protein accumulation Therapeutic vaccines Thylakoids Translational fusion Recombinant proteins beta glucuronidase hybrid protein oncogene protein E7, Human papillomavirus type 16 protein E7 chloroplast genetics growth, development and aging metabolism molecular cloning phenotype thylakoid tobacco transgenic plant Chloroplasts Cloning, Molecular Glucuronidase Papillomavirus E7 Proteins Phenotype Plants, Genetically Modified Recombinant Fusion Proteins Thylakoids Tobacco Human papillomavirus (HPV) is the causal agent of cervical cancer, one of the most common causes of death in women worldwide, and its E7 antigen is the major candidate for a therapeutic vaccine. The large scale production of E7 by molecular farming that would lead to the development of a safe and inexpensive vaccine is impaired by its low accumulation level in the plant cell. To enhance antigen production in the plastids, two alternative strategies were carried out: the expression of E7 as a translational fusion to β-glucuronidase enzyme and redirection of E7 into the thylakoid lumen. The use of the β-glucuronidase as a partner protein turned out to be a successful strategy, antigen expression levels were enhanced between 30 and 40 times relative to unfused E7. Moreover, best accumulation, albeit at a high metabolic cost that compromised biomass production, was obtained redirecting E7 into the thylakoid lumen by the incorporation of the N-terminal transit peptide, Str. Following this approach lumenal E7 production exceeded the stromal by two orders of magnitude. Our results highlight the relevance of exploring different strategies to improve recombinant protein stability for certain transgenes in order to exploit potential advantages of recombinant protein accumulation in chloroplasts. © 2014, Springer Science+Business Media New York. Fil:Morgenfeld, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Lentz, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Segretin, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2014 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10736085_v56_n11_p1021_Morgenfeld http://hdl.handle.net/20.500.12110/paper_10736085_v56_n11_p1021_Morgenfeld |
institution |
Universidad de Buenos Aires |
institution_str |
I-28 |
repository_str |
R-134 |
collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
topic |
E7 antigen Fusion protein Human papillomavirus Molecular farming Thylakoid translocation Transplastomic tobacco Antigens Oncogenic viruses Plants (botany) Tobacco Vaccines Fusion proteins Human papillomavirus Large scale productions Molecular farming Protein accumulation Therapeutic vaccines Thylakoids Translational fusion Recombinant proteins beta glucuronidase hybrid protein oncogene protein E7, Human papillomavirus type 16 protein E7 chloroplast genetics growth, development and aging metabolism molecular cloning phenotype thylakoid tobacco transgenic plant Chloroplasts Cloning, Molecular Glucuronidase Papillomavirus E7 Proteins Phenotype Plants, Genetically Modified Recombinant Fusion Proteins Thylakoids Tobacco |
spellingShingle |
E7 antigen Fusion protein Human papillomavirus Molecular farming Thylakoid translocation Transplastomic tobacco Antigens Oncogenic viruses Plants (botany) Tobacco Vaccines Fusion proteins Human papillomavirus Large scale productions Molecular farming Protein accumulation Therapeutic vaccines Thylakoids Translational fusion Recombinant proteins beta glucuronidase hybrid protein oncogene protein E7, Human papillomavirus type 16 protein E7 chloroplast genetics growth, development and aging metabolism molecular cloning phenotype thylakoid tobacco transgenic plant Chloroplasts Cloning, Molecular Glucuronidase Papillomavirus E7 Proteins Phenotype Plants, Genetically Modified Recombinant Fusion Proteins Thylakoids Tobacco Morgenfeld, Mauro Miguel Lentz, Ezequiel Matías Segretin, María Eugenia Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
topic_facet |
E7 antigen Fusion protein Human papillomavirus Molecular farming Thylakoid translocation Transplastomic tobacco Antigens Oncogenic viruses Plants (botany) Tobacco Vaccines Fusion proteins Human papillomavirus Large scale productions Molecular farming Protein accumulation Therapeutic vaccines Thylakoids Translational fusion Recombinant proteins beta glucuronidase hybrid protein oncogene protein E7, Human papillomavirus type 16 protein E7 chloroplast genetics growth, development and aging metabolism molecular cloning phenotype thylakoid tobacco transgenic plant Chloroplasts Cloning, Molecular Glucuronidase Papillomavirus E7 Proteins Phenotype Plants, Genetically Modified Recombinant Fusion Proteins Thylakoids Tobacco |
description |
Human papillomavirus (HPV) is the causal agent of cervical cancer, one of the most common causes of death in women worldwide, and its E7 antigen is the major candidate for a therapeutic vaccine. The large scale production of E7 by molecular farming that would lead to the development of a safe and inexpensive vaccine is impaired by its low accumulation level in the plant cell. To enhance antigen production in the plastids, two alternative strategies were carried out: the expression of E7 as a translational fusion to β-glucuronidase enzyme and redirection of E7 into the thylakoid lumen. The use of the β-glucuronidase as a partner protein turned out to be a successful strategy, antigen expression levels were enhanced between 30 and 40 times relative to unfused E7. Moreover, best accumulation, albeit at a high metabolic cost that compromised biomass production, was obtained redirecting E7 into the thylakoid lumen by the incorporation of the N-terminal transit peptide, Str. Following this approach lumenal E7 production exceeded the stromal by two orders of magnitude. Our results highlight the relevance of exploring different strategies to improve recombinant protein stability for certain transgenes in order to exploit potential advantages of recombinant protein accumulation in chloroplasts. © 2014, Springer Science+Business Media New York. |
author |
Morgenfeld, Mauro Miguel Lentz, Ezequiel Matías Segretin, María Eugenia |
author_facet |
Morgenfeld, Mauro Miguel Lentz, Ezequiel Matías Segretin, María Eugenia |
author_sort |
Morgenfeld, Mauro Miguel |
title |
Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
title_short |
Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
title_full |
Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
title_fullStr |
Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
title_full_unstemmed |
Translational Fusion and Redirection to Thylakoid Lumen as Strategies to Enhance Accumulation of Human Papillomavirus E7 Antigen in Tobacco Chloroplasts |
title_sort |
translational fusion and redirection to thylakoid lumen as strategies to enhance accumulation of human papillomavirus e7 antigen in tobacco chloroplasts |
publishDate |
2014 |
url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10736085_v56_n11_p1021_Morgenfeld http://hdl.handle.net/20.500.12110/paper_10736085_v56_n11_p1021_Morgenfeld |
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1768545842459836416 |