Transient Expression of Foot and Mouth Disease Virus (FMDV) Coat Protein in Tobacco (Nicotiana tabacom) via Agroinfiltration

Authors

  • Hasan Marashi Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN
  • Nasrin Moshtaghi Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN
  • Saeid Malekzadeh-Shafaroudi Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN
  • Saeid Zibaee Razi Vaccine and Serum Research Institute, Mashhad, I.R. IRAN
Abstract:

Background: Transient and stable transformation of host plants are the common techniques to produce transgenic plants. However, the main drawback of stable transformation is the fact that it takes quite a long time to produce a transgenic line. While, transient gene expression is a quick method to produce recombinant proteins in plants.  Objective: The main goal of the present study was to evaluate efficient agroinfiltration as an efficient and rapid method for production of recombinant antigen of FMDV. Materials and Methods: Tobacco leaves were transformed via agroinfiltration using a needle-free syringe. Presence of the gene cassette was verified by polymerase chain reaction (PCR). Expression of the foreign gene was evaluated using Real Time PCR, protein dot blot and enzyme-linked immunosorbent assay (ELISA). Results: PCR analysis confirmed successful transformation of plant leaves. Expression of foreign protein was confirmed at both transcription and translation levels. Results of Real Time PCR assay indicated that the foreign gene was transcribed in transformed leaves. ELISA results showed that the foreign gene was expressed in the transformed leaves in high level. Conclusion: Here, the efficacy of agroinfiltration for transient expression of FMDV coat protein in tobacco was illustrated. Accordingly, transient agroinfilteration expedites the process of recombinant antigens expression in plant tissues.

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Journal title

volume 12  issue 3

pages  28- 34

publication date 2014-10-01

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