Change in the Basic Structure of the Rabies Virus Glycoprotein by Reverse Genetics

Miandehi, Narges; bidoki, kazem; Ajorloo, Mehdi; Gholami, Alireza

doi:10.30699/ijmm.14.4.348

year 14, Issue 4 (July - August 2020) Iran J Med Microbiol 2020, 14(4): 348-360 | Back to browse issues page

‎ 10.30699/ijmm.14.4.348

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Miandehi N, bidoki K, Ajorloo M, Gholami A. Change in the Basic Structure of the Rabies Virus Glycoprotein by Reverse Genetics. Iran J Med Microbiol 2020; 14 (4) :348-360
URL: http://ijmm.ir/article-1-1098-en.html

Change in the Basic Structure of the Rabies Virus Glycoprotein by Reverse Genetics

Narges Miandehi¹

, Kazem Bidoki²

, Mehdi Ajorloo³

, Alireza Gholami

⁴

1- Department of Biology, Payam-e Noor University, Tehran, Iran
2- Department of Biology, Payame Noor University, Tehran, Iran
3- Department of Clinical Laboratory Sciences, School of Allied Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
4- The National Center for Reference and Research on Rabies, Virology Department, Pasteur Institute of Iran, Tehran, Iran , a.gholami@pasteur.ac.ir

Abstract: (3903 Views)

Background: Rabies is a deadly zoonotic disease that is caused by the rabies virus. The virus can infect and disrupt the central nervous system of a rabid patient. The rabies virus is a neurotropic single stranded RNA virus. Glycoprotein (G) is the most important protein that binds to the cellular receptors and also induces an immune response against the virus in the host. Using reverse genetics technology, the glycoprotein gene could be modified and a virus with higher immunogenicity or lower pathogenicity.
Materials & Methods: In this study, we designed a mutation in the sequence of glycoprotein gene using a software, on the main antigenic site II of the Pasteur virus strain at the position of 42-34 amino acids. Agene fragment in the cloning vector containing the rabies virus genome was replaced by the synthesized construct containing the altered gene by two restricted enzymes, and then cloned. The T7-BHK cell under the T7 phage promoter control was transfected to express the glycoprotein gene, along with the construct and vectors expressing the N, P, and L genes of the rabies virus as well as the full genome. After expressing and confirming viral genes, it was cultured and amplified in BSR cell.
Results: after cloning and expression of the recombinant virus in the target cell, the vector containing the mutated gene led to the rescue of the recombinant virus. The recombinant virus cultured and propagated in the BSR cells, then the genome was extracted and finally confirmed by sequencing.
Conclusion: The rescued recombinant virus can be used for research studies or in the vaccines manufacturing, provide that the antigenicity is maintained or increased.

Keywords: Rabies, Reverse genetics, Vaccine, Glycoprotein, Recombinant virus

Full-Text [PDF 1382 kb] (1466 Downloads) | | Full-Text (HTML) (1166 Views)

Type of Study: Original Research Article | Subject: Medical Virology
Received: 2020/04/5 | Accepted: 2020/06/2 | ePublished: 2020/07/20

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