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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/23071

Title: Expression of the Vibrio cholerae toxin B subunit in the live recombinant vector Salmonella typhi Ty21a
Authors: Aguila, Antonio La O.
Puerta, Ana Herrera
Fernández, Pedro Estévez
Martínez Benítez, Máximo
Aguilera, César Gómez
Imía, Luis García
Domínguez, Armando Acosta
Keywords: CTB, IPTG induction, protein expression, Salmonella typhi Ty21a, vaccine vectors
CTB, expresión de proteínas, inducción con IPTG, Salmonella typhi Ty21a, vectores vacunales
Issue Date: 31-Dec-2000
Publisher: Elfos Scientiae
Citation: Biotecnologia Aplicada (ISSN: 0684-4551) Vol 17 Num 1
Abstract: The strategies to develop effective bacterial vectors have mainly relied on the genetic manipulation of selected microorganisms to ensure the correct presentation to the immune system of convenient amounts of foreign antigens. A new approach consisting in the oral delivery of bacterial vectors carrying adequate quantities of foreign antigens previously expressed in vitro, has been developed. In vitro expression of the cholera toxin binding subunit (CTB) by exogenous induction was studied in the recombinant vector Salmonella typhi   Ty21a, in order to create an experimental tool to further evaluate such an approach. Parameters related to the effectiveness of immunization such as plasmid stability, bacterial viability, and timing of induction, were evaluated as elsewhere, while CTB expression and intracellular location were assayed by GM1-ELISA in samples harvested hourly from cultures induced at 2, 3, 4, and 5 h after inoculation, and were grown for six more hours. CTB expression kinetics was significantly dependent on the timing of induction. The maximal average activity-800 ng/mL of culture-was reached between 2 and 3 h after induction and a lower value of 750 ng/mL remained steady. Seventy percent of the total activity was periplasmic. No differences were detected between induced and control cultures, but in antigen expression. These results are of practical value for the experimental testing of the suggested approach. CTB, IPTG induction, protein expression, Salmonella typhi Ty21a, vaccine vectors
URI: http://hdl.handle.net/1807/23071
Other Identifiers: http://www.bioline.org.br/abstract?id=ba00003
Rights: Copyright 2000 Elfos Scientiae
Appears in Collections:Bioline International Legacy Collection

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