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

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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.

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Authors and Affiliations

  1. División de Vacunas, Centro de Ingeniería Genética y Biotecnología, 10600, La Habana, Cuba

    Iris Valdés, Lisset Hermida, Aída Zulueta, Jorge Martín, Ricardo Silva & Gerardo Guillen

  2. Departamento de Virología, Instituto de Medicina Tropical “Pedro Kourí, Centro Colaborador OPS/OMS en Enfermedades Virales, La Habana, Cuba

    Mayling Álvarez & María G. Guzman

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  1. Iris Valdés
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  2. Lisset Hermida
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  3. Aída Zulueta
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  4. Jorge Martín
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  5. Ricardo Silva
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  6. Mayling Álvarez
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  7. María G. Guzman
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  8. Gerardo Guillen
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Correspondence to Iris Valdés.

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Valdés, I., Hermida, L., Zulueta, A. et al. Expression in Pichia pastoris and immunological evaluation of a truncated dengue envelope protein. Mol Biotechnol 35, 23–30 (2007). https://doi.org/10.1385/MB:35:1:23

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