Journal of Kerman University of Medical Sciences

Production of HPV16-L1 through BL21/pET32a expression system

Document Type : Original Article

Authors

Sanaz Jahandideh 1
Emad Behboudi 2
Hadi Razavi-Nikoo 3
Abdolvahab Moradi 3

1 Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran

2 2 Department of Basic Sciences, Khoy University of Medical Sciences, Khoy, Iran

3 Department of Microbiology, Golestan University of Medical Sciences

https://doi.org/10.34172/jkmu.2023.39
Abstract
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.

Highlights

Sanaz Jahandideh(Google scholar)(Pubmed)

Emad Behboudi(Google scholar)(Pubmed)

Hadi Razavi-Nikoo(Google scholar)(Pubmed)

Abdolvahab Moradiv(Google scholar)(Pubmed)

Keywords

HPV
VLP
Vaccine
BL21
pET32
Subjects
Journal of Kerman University of Medical Sciences
Volume 30, Issue 4
July and August 2023
Pages 233-237

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