Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter

dc.creatorMariz, F. C.
dc.creatorCoimbra, E. C.
dc.creatorJesus, A. L. S.
dc.creatorNascimento, L. M.
dc.creatorTorres, F. A. G.
dc.creatorFreitas, A. C.
dc.date2017-11-16T18:12:49Z
dc.date2017-11-16T18:12:49Z
dc.date2015
dc.date.accessioned2023-09-26T23:25:07Z
dc.date.available2023-09-26T23:25:07Z
dc.identifierMARIZ, 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, v. 2015, p. 594120, 2015.
dc.identifier2314-6141
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/23244
dc.identifier10.1155/2015/594120
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8890363
dc.descriptionThe 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 (PPGK1) from the methylotrophic yeast Pichia pastoris. The L1 gene was cloned under regulation of PPGK1 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.
dc.formatapplication/pdf
dc.languagepor
dc.rightsopen access
dc.subjectBiotecnologia
dc.subjectProteínas do Capsídeo
dc.subjectProteínas Oncogênicas Virais
dc.subjectNeoplasias do Colo do Útero
dc.subjectVacinas de Partículas Semelhantes a Vírus
dc.subjectBiotechnology
dc.subjectCapsid Proteins
dc.subjectFemale
dc.subjectGene Expression Regulation, Viral
dc.subjectHuman papillomavirus 16
dc.subjectHumans
dc.subjectOncogene Proteins, Viral
dc.subjectPichia
dc.subjectPromoter Regions, Genetic
dc.subjectUterine Cervical Neoplasms
dc.subjectVaccines, Virus-Like Particle
dc.subjectBiotecnologia
dc.subjectProteínas do Capsídeo
dc.subjectimunologia
dc.subjectProteínas Oncogênicas Virais
dc.subjectimunologia
dc.subjectNeoplasias do Colo do Útero
dc.subjectprevenção & controle
dc.subjectVacinas de Partículas Semelhantes a Vírus
dc.subjectFeminino
dc.subjectRegulação Viral da Expressão Gênica
dc.subjectPapillomavirus Humano 16
dc.subjectpatogenicidade
dc.subjectHumanos
dc.subjectquímica
dc.subjectPichia
dc.subjectRegiões Promotoras Genéticas
dc.subjectvirologia
dc.titleDevelopment of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter
dc.typeArticle


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