Molecular Biology Reports

, Volume 45, Issue 6, pp 2237–2246 | Cite as

Constructing and transient expression of a gene cassette containing edible vaccine elements and shigellosis, anthrax and cholera recombinant antigens in tomato

  • Jafari Davod
  • Dehghan Nayeri FatemehEmail author
  • Hossein Honari
  • Ramin Hosseini
Original Article


Shigella dysenteriae causing shigellosis is one of the diseases that threaten the health of human society in the developing countries. In Shigella, IpaD gene is one of the key pathogenic genes causing strong mucosal immune system reactions. Anthrax disease is caused by Bacillus anthracis. PA protective antigen is one of the subunits in anthrax toxin complex responsible for the transfer of other subunits into the cytosol of host cells. The 20 kDa subunit of PA (PA20) has the property of immunogenicity. CTxB or B subunit of Vibrio cholerae toxin (CT) is a non-toxic protein and has the function to transfer toxic subunit into cytosol of the host cells by binding to GM1 receptor. The aim of this study was to fuse PA20, ipaD and CTxB and transform tomato plants by this cassette in order to produce an oral vaccine against shigellosis, anthrax and cholera. CTxB was used for these two antigens as an immune adjuvant. IpaD and PA20 genes were cloned in pBI121 containing the CTxB gene and Extensin signal peptide. In order to evaluate the transient expression of Shigellosis, Anthrax and Cholera antigens, agro-infiltrated tomato tissues were inoculated with Agrobacterium tumefaciens containing the gene cassette. Cloning was confirmed by PCR, enzymatic digestion and sequencing techniques. Expression of the antigens was examined by SDS-PAGE, dot blot and ELISA. Maturate green fruits demonstrated the highest expression of the recombinant proteins. The first phase of this study was carried out for cloning and expressing of CtxB, ipaD and PA20 antigens in tomato. In the next phase, we aim to analyze the immunogenicity of this vaccine candidate in laboratory animals.


Anthrax Cholera Tomato Edible vaccine Shigellosis Agro-infiltration 



This work was done in Agricultural Biotechnology Department of Imam Khomeini International University. We appreciate all staffs on their good collaborations.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

Ethical approval and informed consent were not required for this type of study.

Research involving human participants or animals

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Jafari Davod
    • 1
    • 2
  • Dehghan Nayeri Fatemeh
    • 2
    Email author
  • Hossein Honari
    • 3
  • Ramin Hosseini
    • 2
  1. 1.Medical Biotechnology Department, School of Allied MedicineIran University of Medical SciencesTehranIran
  2. 2.Biotechnology Department, Faculty of Agricultural and Natural SciencesImam Khomeini International University (IKIU)QazvinIran
  3. 3.Faculty of Basic ScienceImam Hussein UniversityTehranIran

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