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
Publish place: Journal of Cell and Molecular Research، Vol: 7، Issue: 1
Publish Year: 1394
نوع سند: مقاله ژورنالی
زبان: Persian
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شناسه ملی سند علمی:
JR_JCMR-7-1_007
تاریخ نمایه سازی: 7 بهمن 1395
Abstract:
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 inhairy 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 ofAgrobacterium rhizogenes. Hairy roots developed on MS medium containing 50 mg/L kanamycin and 30 mg/Lmeropenem. 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
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Authors
Amir Ghaffar Shahriari
Department of Plant Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, Iran
Abdol Reza Bagheri
Department of Plant Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, Iran
Mohammad Reza Bassami
Biotechnology Research Group, Ferdowsi University of Mashhad, Mashhad, Iran
Saeed Malek Zadeh Shafaroudi
Department of Plant Biotechnology and Breeding, Ferdowsi University of Mashhad, Mashhad, Iran