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Volume 8, Issue 1 (6-2021)
vacres 2021, 8(1): 67-72 |
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Abedi Kiasari B, Najafi H. Production of Polyomavirus and Herpesvirus Recombinant Glycoproteins with Immunoreactivity Using a Rapid and Novel Expression System in Insect Cells for Applications in Vaccines and Serological Assays. vacres 2021; 8 (1) :67-72
URL: http://vacres.pasteur.ac.ir/article-1-278-en.html
URL: http://vacres.pasteur.ac.ir/article-1-278-en.html
Virology Department, The Faculty of Veterinary Medicine, The University of Tehran, Tehran, Iran
Abstract: (1416 Views)
Introduction: Although, conventional methods for the expression of polyomaviruses and herpesviruses recombinant proteins for serological assays and vaccine developments in baculoviruses are well established, the manipulations are laborious and time consuming.
Methods: A new expression system based on plasmid was used to express two polyomaviruses major capsid protein VP1 (JCV VP1 and BKV VP1), and two herpesviruses glycoproteins (HSV-1 gD and VZV gE) in insect cells. A ligation independent cloning (LIC) was applied to generate the recombinant plasmids. Transfection of Sf9 insect cells were performed using the recombinants. The produced proteins were analysed using SDS-PAGE, immunofluorescence, and immunoblotting. Results: JCV-VP1, BKV-VP1, VZV-gE and HSV-1gD were successfully expressed in the insect cells, 48 h post-infection and detected in cytoplasm and cell membranes with immunoreactivity. This plasmid based expression system took 5 days to express the protein. Conclusion: The plasmid based expression system in insect cells was highly efficient and would be ideal for rapid expression of polyomaviruses and herpesviruses proteins in insect cells to be potentially used in applications such as vaccine components and serological assays.
Methods: A new expression system based on plasmid was used to express two polyomaviruses major capsid protein VP1 (JCV VP1 and BKV VP1), and two herpesviruses glycoproteins (HSV-1 gD and VZV gE) in insect cells. A ligation independent cloning (LIC) was applied to generate the recombinant plasmids. Transfection of Sf9 insect cells were performed using the recombinants. The produced proteins were analysed using SDS-PAGE, immunofluorescence, and immunoblotting. Results: JCV-VP1, BKV-VP1, VZV-gE and HSV-1gD were successfully expressed in the insect cells, 48 h post-infection and detected in cytoplasm and cell membranes with immunoreactivity. This plasmid based expression system took 5 days to express the protein. Conclusion: The plasmid based expression system in insect cells was highly efficient and would be ideal for rapid expression of polyomaviruses and herpesviruses proteins in insect cells to be potentially used in applications such as vaccine components and serological assays.
Keywords: Polyomavirus, Herpes simplex virus, Varicella Zoster virus, Baculovirus expression system, Plasmid based Expression system, Recombinant protein.
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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.