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Expression of Cholera toxin B subunit in Banana callus culture

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Author(s): G. Renuga | R. Saravanan | A. Babu Thandapani | K.R. Arumugam

Journal: Journal of Pharmaceutical Sciences and Research
ISSN 0975-1459

Volume: 2;
Issue: 1;
Start page: 26;
Date: 2010;
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Keywords: Antigen | CT-B | Robusta sp | Transgenic plants | Vibrio cholerae

ABSTRACT
Plants present a low cost agriculturally based effective production system for high value therapeutic system. The process of developing an edible vaccine begins by selecting a suitable vaccine gene of a pathogen and selection of plants to introduce vaccine gene in host. In the present study an attempt was made to produce edible plant vaccineagainst cholera expressed in Banana callus culture. Vibrio cholerae producing cholera toxins (CT) consists of two A (CT-A) and B subunits (CT-B), One subunit has 27 kDa (A subunit) which contain a toxic ADP ribosyl transferase activity and a pentamer of 11.6 kDa (B subunit). The CT-B subunit of Vibrio cholerae cells constituted an oralvaccine against cholera. In order to express CT-B protein in plant system, gene encoding cholera toxin B subunit (CT-B) was cloned into a vector pCAMBIA. CT-B gene was amplified by PCR and cloned into vectors pCAMBIA containing the strong, constitutive 35S CaMV promoter and a reiterated 35S enhancer. The plasmids were transformed into Banana callus via Agrobacterium tumefaciens. Production of CT-B vaccine in banana plant callus for expression and delivery of edible vaccines. Banana callus proteins were analyzed for the confirmation of recombinant CT-B. Transgenic plants expressing CT-B showed the presence of a protein that was recognized by mouse anti CT-B antibody and which migrated to the same position in SDS-PAGE as CT-B derived from Vibrio cholerae. Edible vaccines against Vibrio cholerae has expressed in Banana which could be consumed raw in the form of fruit.
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