| Autor: |
Sanaz Jahandideh, Emad Behboudi, Hadi Razavi-Nikoo, Abdolvahab Moradi |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Journal of Kerman University of Medical Sciences, Vol 30, Iss 4, Pp 233-237 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2008-2843 |
| DOI: |
10.34172/jkmu.2023.39 |
| Popis: |
Introduction: The Human papillomaviruses (HPV) main capsid protein L1 is naturally capable to self-assemble as virus-like particles (VLPs). There are different recombinant protein expression systems such as bacteria, yeast, insect, plant, and mammalian cells for generation of VLP-based candidate vaccines targeting various pathogens. In this study, we produced HPV-L1 protein by BL21/pET32a expression system and VLP production was confirmed.Material & Method: The recombinant plasmid pET32/L1 was transformed into Escherichia. coli BL21 and selected with ampicillin. The positive clones containing the recombinant plasmid pET32/L1 were assessed by restriction endonucleases HindIII and XhoI and sequence analysis. The expression of HPV16-L1 fusion protein in E. coli BL21was identified by SDS-PAGE and Western blotting. VLPs were evaluated using electron microscopy.Result: A codon-optimized L1 gene was expressed in BL21 under the control of the T7/lac promoter. Purification of L1 protein was achieved after Ni NTA chromatography. The 60kDa protein was detected in the lysates of BL21, recognized as HPV16- L1 protein by Western blotting. VLPs were confirmed using electron microscopy.Conclusion: In this study, we established a high-efficient recombinant E. coli expression system for the production of HPV 16- L1 protein. The generated L1 protein was correctly self-assembled into VLPs. Therefore, BL21/pET32a as a prokaryotic expression system is a potent tool for HPV16-L1 VLP production. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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