Expression of the B subunit of Escherichia coli heat-labile enterotoxin as a fusion protein in transgenic tomato

Abstract

Epitopes often require co-delivery with an adjuvant or targeting protein to enable recognition by the immune system. This paper reports the ability of transgenic tomato plants to express a fusion protein consisting of the B subunit of the Escherichia coli heat-labile enterotoxin (LTB) and an immunocontraceptive epitope. The fusion protein was found to assemble into pentamers, as evidenced by its ability to bind to gangliosides, and had an average expression level of 37.8 microg g(-1) in freeze-dried transgenic tissues. Processing of selected transgenic fruit resulted in a 16-fold increase in concentration of the antigen with minimal loss in detectable antigen. The species-specific nature of this epitope was shown by the inability of antibodies raised against non-target species to detect the LTB fusion protein. The immunocontraceptive ability of this vaccine will be tested in future pilot mice studies.

Expression of Escherichia coli heat-labile enterotoxin B subunit in transgenic tomato (Solanum lycopersicum L.) fruit

“…Based on the results of the In other study, we found that the amount of LTB protein of in vitro transgenic tomato leaves was approximately 0.8-1.0% of TSPs (Nhi et al 2010). Walmsley et al (2003) found that the LTB protein of transgenic tomato fruit was assembled into pentamers. It showed an average expression level of 37.8 μg/g in freeze-dried transgenic tissues.…”

Production of vaccines and vaccine components in corn

“…Varying lengths of proteins have been fused to LT-B and CT-B with or without a hinge and expressed in both bacteria and plant systems (Lipscombe et al, 1991, Smerdou et al, 1996, Luci et al, 2006, Walmsley et al, 2003. However, success of the fusion protein approach has to be evaluated on a case-by-case basis (Rigano et al, 2003).…”

“…A number of studies were carried out involving expression of LT-B and CT-B fusions with unrelated antigens in plants (Kim et al, 2004a, Kim et al 2004b, Kim et al, 2004c, Kang et al, 2006, Walmsley et al, 2003 The present work aimed at utilizing LT-B and CT-B as carriers for a human protein 'amyloid beta' and its epitopes to generate an orally administered plant-derived vaccine against Alzheimer's disease (AD). Alzheimer's disease is a neurodegenerative disorder characterized by cognitive decline, memory loss, language deficits and personality changes.…”

“…Hence, oral immunization with CT-B and LT-B results in strong humoral and mucosal immune response (Rigano et al, 2003).…”

Estudio transcriptómico y metabolómico del desarrollo partenocárpico del fruto de tomate y aplicaciones biotecnológicas

“…Algunos ejemplos en la literatura demuestran el gran potencial biotecnológico de los frutos transgénicos para la mejora de caracteres agronómicos (Carmi et al, 2003;Ficcadenti et al, 1999), valor nutricional (Mehta et al, 2002;Muir et al, 2001), rasgos organolépticos (Davidovich-Rikanati et al, 2007), e incluso valor terapéutico (Jani et al, 2002;Ma et al, 2003;Ramirez et al, 2007;Walmsley et al, 2003). A pesar de estos casos, las herramientas disponibles para la modificación genética resultan ser bastante rudimentarias, basándose en la mayoría de los casos en unos pocos promotores de fruto bien caracterizados.…”