Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin

dc.creatorNascimento, D. V.
dc.creatorLemes, E. M. B.
dc.creatorQueiroz, J. L. S.
dc.creatorSilva Jr., J. G.
dc.creatorNascimento, H. J.
dc.creatorSilva, E. D.
dc.creatorHirata Jr., R.
dc.creatorDias, A. A. S. O.,
dc.creatorSantos, C. S.
dc.creatorPereira, G. M. B.
dc.creatorGuaraldi, A. L. Mattos
dc.creatorArmoa, G. R. G.
dc.date2017-10-19T10:49:28Z
dc.date2017-10-19T10:49:28Z
dc.date2010
dc.date.accessioned2023-09-26T20:10:35Z
dc.date.available2023-09-26T20:10:35Z
dc.identifierNASCIMENTO, D. V. et al. Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin. Brazilian Journal of Medical and Biological Research, v.43, p.460-466, 2010.
dc.identifier0100-879X
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/22876
dc.identifier1414-431X)
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8850485
dc.descriptionThe construction of a hexahistidine-tagged version of the B fragment of diphtheria toxin (DTB) represents an important step in the study of the biological properties of DTB because it will permit the production of pure recombinant DTB (rDTB) in less time and with higher yields than currently available. In the present study, the genomic DNA of the Corynebacterium diphtheriae Park Williams 8 (PW8) vaccine strain was used as a template for PCR amplification of the dtb gene. After amplification, the dtb gene was cloned and expressed in competent Escherichia coli M15™ cells using the expression vector pQE-30™. The lysate obtained from transformed E. coli cells containing the rDTBPW8 was clarified by centrifugation and purified by affinity chromatography. The homogeneity of the purified rDTBPW8 was confirmed by immunoblotting using mouse polyclonal anti-diphtheria toxoid antibodies and the immune response induced in animals with rDTBPW8 was evaluated by ELISA and dermonecrotic neutralization assays. The main result of the present study was an alternative and accessible method for the expression and purification of immunogenically reactive rDTBPW8 using commercially available systems. Data also provided preliminary evidence that rabbits immunized with rDTBPW8 are able to mount a neutralizing response against the challenge with toxigenic C. diphtheriae
dc.formatapplication/pdf
dc.languageeng
dc.publisherAssociação Brasileira de Divulgação Científica
dc.rightsopen access
dc.subjectDifteria
dc.subjectEscherichia coli
dc.subjectExpressão Gênica
dc.subjectAfinidade do metal imobilizado
dc.subjectFragment B
dc.subjectDiphtheria
dc.subjectdtb gene
dc.subjectE. coli gene expression
dc.subjectImmobilized metal affinity
dc.titleExpression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin
dc.typeArticle


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