Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco
Background: Newcastle disease is an important avian infectious disease that brings about vast economic damage for poultry industry. Transgenic plants represent a cost-effective system for the production of therapeutic proteins and are widely used for the production of poultry vaccines. In an attempt...
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doaj-ee31ca33b953442bb9e79fd91cb372ea2020-11-24T22:38:54ZengElsevierElectronic Journal of Biotechnology0717-34582016-07-0122C384310.1016/j.ejbt.2016.05.003Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobaccoAmir Ghaffar Shahriari0Abdolreza Bagheri1Mohammad Reza Bassami2Saeid Malekzadeh-Shafaroudi3Alireza Afsharifar4Ali Niazi5Ferdowsi University of Mashhad, Department of Crop Biotechnology and Breeding, Mashhad, IranFerdowsi University of Mashhad, Department of Crop Biotechnology and Breeding, Mashhad, IranDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, IranFerdowsi University of Mashhad, Department of Crop Biotechnology and Breeding, Mashhad, IranPlant Virology Research Centre, College of Agriculture, Shiraz University, Shiraz, IranFaculty member of Biotechnology Institute, Biotechnology Institute, Shiraz University, Shiraz, IranBackground: Newcastle disease is an important avian infectious disease that brings about vast economic damage for poultry industry. Transgenic plants represent a cost-effective system for the production of therapeutic proteins and are widely used for the production of poultry vaccines. In an attempt to develop a recombinant vaccine, a plant expression binary vector pBI121, containing the genes encoding Hemagglutinin–Neuraminidase (HN) and Fusion (F) epitopes of Newcastle Disease Virus (NDV) under the control of CaMV35S promoter and NOS terminator was constructed and introduced into the tobacco (Nicotiana tabacum) plant by Agrobacterium-mediated transformation. Results: Putative transgenic plants were screened in a selection medium containing 50 mg/L kanamycin and 30 mg/L meropenem. Integration of the foreign gene in plant genome was confirmed by PCR. Expression of foreign gene was analyzed at transcription level by RT-PCR and at translation level by means of dot blotting and ELISA. All analyses confirmed the expression of recombinant protein. Conclusion: Developments in genetic engineering have led to plant-based systems for recombinant vaccine production. In this research, tobacco plant was used to express F and HN epitopes of NDV. Our results indicate that for the production of recombinant vaccine, it is a novel strategy to use concatenated epitopes without their genetic fusion onto larger scaffold structure such as viral coat protein.http://www.sciencedirect.com/science/article/pii/S0717345816300379Agrobacterium tumefaciensELISARecombinant vaccineTransgenic plants |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeid Malekzadeh-Shafaroudi Alireza Afsharifar Ali Niazi |
spellingShingle |
Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeid Malekzadeh-Shafaroudi Alireza Afsharifar Ali Niazi Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco Electronic Journal of Biotechnology Agrobacterium tumefaciens ELISA Recombinant vaccine Transgenic plants |
author_facet |
Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeid Malekzadeh-Shafaroudi Alireza Afsharifar Ali Niazi |
author_sort |
Amir Ghaffar Shahriari |
title |
Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco |
title_short |
Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco |
title_full |
Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco |
title_fullStr |
Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco |
title_full_unstemmed |
Expression of Hemagglutinin–Neuraminidase and fusion epitopes of Newcastle Disease Virus in transgenic tobacco |
title_sort |
expression of hemagglutinin–neuraminidase and fusion epitopes of newcastle disease virus in transgenic tobacco |
publisher |
Elsevier |
series |
Electronic Journal of Biotechnology |
issn |
0717-3458 |
publishDate |
2016-07-01 |
description |
Background: Newcastle disease is an important avian infectious disease that brings about vast economic damage for poultry industry. Transgenic plants represent a cost-effective system for the production of therapeutic proteins and are widely used for the production of poultry vaccines. In an attempt to develop a recombinant vaccine, a plant expression binary vector pBI121, containing the genes encoding Hemagglutinin–Neuraminidase (HN) and Fusion (F) epitopes of Newcastle Disease Virus (NDV) under the control of CaMV35S promoter and NOS terminator was constructed and introduced into the tobacco (Nicotiana tabacum) plant by Agrobacterium-mediated transformation.
Results: Putative transgenic plants were screened in a selection medium containing 50 mg/L kanamycin and 30 mg/L meropenem. Integration of the foreign gene in plant genome was confirmed by PCR. Expression of foreign gene was analyzed at transcription level by RT-PCR and at translation level by means of dot blotting and ELISA. All analyses confirmed the expression of recombinant protein.
Conclusion: Developments in genetic engineering have led to plant-based systems for recombinant vaccine production. In this research, tobacco plant was used to express F and HN epitopes of NDV. Our results indicate that for the production of recombinant vaccine, it is a novel strategy to use concatenated epitopes without their genetic fusion onto larger scaffold structure such as viral coat protein. |
topic |
Agrobacterium tumefaciens ELISA Recombinant vaccine Transgenic plants |
url |
http://www.sciencedirect.com/science/article/pii/S0717345816300379 |
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