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Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter
Joint Authors
Torres, Fernando Araripe Gonçalves
Freitas, Antonio Carlos de
Mariz, F. C.
Coimbra, E. C.
Jesus, A. L. S.
Nascimento, L. M.
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-05-18
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
The human papillomavirus (HPV) L1 major capsid protein, which forms the basis of the currently available vaccines against cervical cancer, self-assembles into virus-like particles (VLPs) when expressed heterologously.
We report the development of a biotechnology platform for HPV16 L1 protein expression based on the constitutive PGK1 promoter ( P PGK 1 ) from the methylotrophic yeast Pichia pastoris.
The L1 gene was cloned under regulation of P PGK 1 into pPGKΔ3 expression vector to achieve intracellular expression.
In parallel, secretion of the L1 protein was obtained through the use of an alternative vector called pPGKΔ3α, in which a codon optimized α-factor signal sequence was inserted.
We devised a work-flow based on the detection of the L1 protein by dot blot, colony blot, and western blot to classify the positive clones.
Finally, intracellular HPV VLPs assembly was demonstrated for the first time in yeast cells.
This study opens up perspectives for the establishment of an innovative platform for the production of HPV VLPs or other viral antigens for vaccination purposes, based on constitutive expression in P.
pastoris.
American Psychological Association (APA)
Mariz, F. C.& Coimbra, E. C.& Jesus, A. L. S.& Nascimento, L. M.& Torres, Fernando Araripe Gonçalves& Freitas, Antonio Carlos de. 2015. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International،Vol. 2015, no. 2015, pp.1-11.
https://search.emarefa.net/detail/BIM-1056049
Modern Language Association (MLA)
Mariz, F. C.…[et al.]. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International No. 2015 (2015), pp.1-11.
https://search.emarefa.net/detail/BIM-1056049
American Medical Association (AMA)
Mariz, F. C.& Coimbra, E. C.& Jesus, A. L. S.& Nascimento, L. M.& Torres, Fernando Araripe Gonçalves& Freitas, Antonio Carlos de. Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter. BioMed Research International. 2015. Vol. 2015, no. 2015, pp.1-11.
https://search.emarefa.net/detail/BIM-1056049
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1056049