Recombinant Rabies Virus With a Multiple Cloning Site: A Platform Capable of Immunological Alterations | ||
| Immunoregulation | ||
| مقاله 6، دوره 4، شماره 2، بهار 2021، صفحه 101-108 اصل مقاله (1.14 M) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.32598/Immunoregulation.4.2.5 | ||
| نویسندگان | ||
| Seyed Hamidreza Naghi Mousavi1؛ Alireza Gholami* 2؛ Mojtaba Jafarinia1؛ Mehdi Ajorloo* 3 | ||
| 1Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran. | ||
| 2Viral Vaccines Production Unit, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran. | ||
| 3Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran. | ||
| چکیده | ||
| Background: Rabies is fatal encephalitis, i.e., preventable by appropriate vaccination. Reverse genetics has proved promising for manipulating the rabies virus immunological characteristics. The insertion or deletion of a gene from the rabies genome could render specific functions to the rabies virus. Materials and Methods: A multiple cloning site including 111 nucleotides long harboring 10 single-cut restriction sites have been designed. The designed fragment was cloned between the G and L genes of the rabies virus genome. The recombinant rabies virus was rescued, and its infectivity was confirmed in the BHK-21 cell line. The recombinant virus propagation was compared with the initial rabies virus strain. Statistical analysis was performed using GraphPad Prism. Results: The cloning and localization of the multiple cloning site were verified by nucleotide sequencing. The recombinant virus properly propagated and rescued in the BHK-21 cell line. Comparing the recombinant virus with the initial rabies virus has shown that both viruses had similar functionality and propagation rate. Conclusion: The recombinant virus obtained in the present study could facilitate further cloning experiments. Examples include constructing a marker virus, carrying green fluorescent protein to be used either in rabies immunity assays or tracking the virus infection in relevant tissues. | ||
| کلیدواژهها | ||
| Rabies virus؛ Reverse genetics؛ Virus engineering؛ Molecular cloning | ||
| مراجع | ||
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