Among the Dengue virus structural proteins, the Envelope glycoprotein is the most important because of its antigenic characteristics. In this work, the E protein from Dengue-2 virus truncated at the C-terminus region was successfully expressed in Pichia pastoris. The E2trunc gene was cloned under the AOX1 promoter from P. pastoris and the signal peptide of the sucrose invertase gene from Saccharomyces cerevisiae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of expression revealed the presence of a protein with the expected size, which was completely associated to the insoluble fraction after cellular disruption. The recombinant N-glycosylated protein reacted with two conformational antibodies against Dengue-2, indicating a proper folding of it. In addition, it was able to induce antiviral antibodies after mice immunization. © 2006 Humana Press Inc. All rights of any nature whatsoever reserved.

Expression in Pichia pastoris and immunological evaluation of a truncated Dengue envelope protein / I. Valdés, L. Hermida, A. Zulueta, J. Martín, R. Silva, M. Álvarez, M.G. Guzmán, G. Guillén. - In: MOLECULAR BIOTECHNOLOGY. - ISSN 1073-6085. - 35:1(2007), pp. 23-30.

Expression in Pichia pastoris and immunological evaluation of a truncated Dengue envelope protein

A. Zulueta;
2007

Abstract

Among the Dengue virus structural proteins, the Envelope glycoprotein is the most important because of its antigenic characteristics. In this work, the E protein from Dengue-2 virus truncated at the C-terminus region was successfully expressed in Pichia pastoris. The E2trunc gene was cloned under the AOX1 promoter from P. pastoris and the signal peptide of the sucrose invertase gene from Saccharomyces cerevisiae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of expression revealed the presence of a protein with the expected size, which was completely associated to the insoluble fraction after cellular disruption. The recombinant N-glycosylated protein reacted with two conformational antibodies against Dengue-2, indicating a proper folding of it. In addition, it was able to induce antiviral antibodies after mice immunization. © 2006 Humana Press Inc. All rights of any nature whatsoever reserved.
Dengue virus; Expression; Glycosylation; Pichia pastoris yeast; Secretion; Biochemistry, Genetics and Molecular Biology (all); Biochemistry; Biotechnology; Bioengineering; Applied Microbiology and Biotechnology
Settore BIO/10 - Biochimica
Settore BIO/11 - Biologia Molecolare
Settore CHIM/11 - Chimica e Biotecnologia delle Fermentazioni
MOLECULAR BIOTECHNOLOGY
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/491362
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