Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease
Newcastle is a significant avian disease continuing to cause considerable loss. Developments in genetic engineering have led to plant-based platforms for human and animal vaccine production. Recombinant vaccine production in hairy root systems have several advantages over stable expression in whole...
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| Format: | Article |
| Language: | English |
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Ferdowsi University of Mashhad
2015-07-01
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| Series: | Journal of Cell and Molecular Research |
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| Online Access: | https://jcmr.um.ac.ir/article_27898_982c744b93a609751d69888786100db6.pdf |
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| author | Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeed Malekzadeh Shafaroudi Ali Reza Afsharifar |
| author_facet | Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeed Malekzadeh Shafaroudi Ali Reza Afsharifar |
| author_sort | Amir Ghaffar Shahriari |
| collection | DOAJ |
| description | Newcastle is a significant avian disease continuing to cause considerable loss. Developments in genetic engineering have led to plant-based platforms for human and animal vaccine production. Recombinant vaccine production in hairy root systems have several advantages over stable expression in whole plants, including high growth rates, ready genetic manipulations, high levels of recombinant protein production, and the potential for bioreactor culture. In an attempt to develop a recombinant vaccine in hairy roots, the sequences encoding fusion (F) and haemagglutinin-neuraminidase (HN) epitopes of Newcastle disease virus were cloned in pBI121 expression vector which was then transferred into leaf disks of tobacco (Nicotiana tabaccum) 'Turkish' cultivar by means of Agrobacterium rhizogenes. Hairy roots developed on MS medium containing 50 mg/L kanamycin and 30 mg/L meropenem. Incorporation of the heterologous gene in the genome of hairy roots was confirmed by PCR. Expression analyses were performed by real-time PCR at transcription level and by dot-blot and ELISA assays at translation level, all confirming the expression of the heterologous gene and production of the recombinant protein. |
| first_indexed | 2024-04-11T15:22:02Z |
| format | Article |
| id | doaj.art-fcd1f4125e7a4ce891339f304d06edf9 |
| institution | Directory Open Access Journal |
| issn | 2008-9147 2717-3364 |
| language | English |
| last_indexed | 2024-04-11T15:22:02Z |
| publishDate | 2015-07-01 |
| publisher | Ferdowsi University of Mashhad |
| record_format | Article |
| series | Journal of Cell and Molecular Research |
| spelling | doaj.art-fcd1f4125e7a4ce891339f304d06edf92022-12-22T04:16:21ZengFerdowsi University of MashhadJournal of Cell and Molecular Research2008-91472717-33642015-07-0171111810.22067/jcmr.v7i1.4162127898Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle DiseaseAmir Ghaffar Shahriari0Abdolreza Bagheri1Mohammad Reza Bassami2Saeed Malekzadeh Shafaroudi3Ali Reza Afsharifar4Ferdowsi University of MashhadFerdowsi University of MashhadFerdowsi University of MashhadFerdowsi University of MashhadShiraz UniversityNewcastle is a significant avian disease continuing to cause considerable loss. Developments in genetic engineering have led to plant-based platforms for human and animal vaccine production. Recombinant vaccine production in hairy root systems have several advantages over stable expression in whole plants, including high growth rates, ready genetic manipulations, high levels of recombinant protein production, and the potential for bioreactor culture. In an attempt to develop a recombinant vaccine in hairy roots, the sequences encoding fusion (F) and haemagglutinin-neuraminidase (HN) epitopes of Newcastle disease virus were cloned in pBI121 expression vector which was then transferred into leaf disks of tobacco (Nicotiana tabaccum) 'Turkish' cultivar by means of Agrobacterium rhizogenes. Hairy roots developed on MS medium containing 50 mg/L kanamycin and 30 mg/L meropenem. Incorporation of the heterologous gene in the genome of hairy roots was confirmed by PCR. Expression analyses were performed by real-time PCR at transcription level and by dot-blot and ELISA assays at translation level, all confirming the expression of the heterologous gene and production of the recombinant protein.https://jcmr.um.ac.ir/article_27898_982c744b93a609751d69888786100db6.pdfrecombinant vaccineantigen expressionnewcastle diseasehairy rootsagrobacterium rhizogenes |
| spellingShingle | Amir Ghaffar Shahriari Abdolreza Bagheri Mohammad Reza Bassami Saeed Malekzadeh Shafaroudi Ali Reza Afsharifar Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease Journal of Cell and Molecular Research recombinant vaccine antigen expression newcastle disease hairy roots agrobacterium rhizogenes |
| title | Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease |
| title_full | Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease |
| title_fullStr | Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease |
| title_full_unstemmed | Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease |
| title_short | Cloning and Expression of Fusion (F) and Haemagglutinin-neuraminidase (HN) Epitopes in Hairy Roots of Tobacco (Nicotiana tabaccum) as a Step Toward Developing a Candidate Recombinant Vaccine Against Newcastle Disease |
| title_sort | cloning and expression of fusion f and haemagglutinin neuraminidase hn epitopes in hairy roots of tobacco nicotiana tabaccum as a step toward developing a candidate recombinant vaccine against newcastle disease |
| topic | recombinant vaccine antigen expression newcastle disease hairy roots agrobacterium rhizogenes |
| url | https://jcmr.um.ac.ir/article_27898_982c744b93a609751d69888786100db6.pdf |
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