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

مقاله 1، دوره 7، شماره 1 - شماره پیاپی 13، مهر 2015، صفحه 11-18   

نوع مقاله: Research Articles

شناسه دیجیتال (DOI): 10.22067/jcmr.v7i1.41621

نویسندگان

Amir Ghaffar Shahriari^{* 1}؛ Abdolreza Bagheri¹؛ Mohammad Reza Bassami¹؛ Saeed Malekzadeh Shafaroudi¹؛ Ali Reza Afsharifar²

¹Ferdowsi University of Mashhad

²Shiraz University

چکیده

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.

کلیدواژه‌ها

Recombinant vaccine؛ Antigen Expression؛ Newcastle Disease؛ Hairy Roots؛ Agrobacterium Rhizogenes