Sharma, Manoj Kumar and Singh, Nirmal Kumar and Jani, Dewal and Sisodia, Rama and Thungapathra, M and Gautam, J K and Meena, L S and Singh, Yogendra and Ghosh, Amit and Tyagi, Akhilesh Kumar and Sharma, Arun Kumar (2008) Expression of toxin co-regulated pilus subunit A (TCPA) of Vibrio cholerae and its immunogenic epitopes fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum). Plant cell reports, 27 (2). pp. 307-18. ISSN 0721-7714

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Abstract

For protection against cholera, it is important to develop efficient vaccine capable of inducing anti-toxin as well as anti-colonizing immunity against Vibrio cholerae infections. Earlier, expression of cholera toxin B subunit (CTB) in tomato was reported by us. In the present investigation, toxin co-regulated pilus subunit A (TCPA), earlier reported to be an antigen capable of providing anti-colonization immunity, has been expressed in tomato. Further, to generate more potent combinatorial antigens, nucleotides encoding P4 or P6 epitope of TCPA were fused to cholera toxin B subunit gene (ctxB) and expressed in tomato. Presence of transgenes in the tomato genome was confirmed by PCR and expression of genes was confirmed at transcript and protein level. TCPA, chimeric CTB-P4 and CTB-P6 proteins were also expressed in E. coli. TCPA protein expressed in E. coli was purified to generate anti-TCPA antibodies in rabbit. Immunoblot and G(M1)-ELISA verified the synthesis and assembly of pentameric chimeric proteins in fruit tissue of transgenic tomato plants. The chimeric protein CTB-P4 and CTB-P6 accumulated up to 0.17 and 0.096% of total soluble protein (TSP), respectively, in tomato fruits. Whereas expression of TCPA, CTB-P4 and CTB-P6 in E. coli can be utilized for development of conventional vaccine, expression of these antigens which can provide both anti-toxin as well as anti-colonization immunity, has been demonstrated in plants, in a form which is potentially capable of inducing immune response against cholera infection.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Springer Science.
Subjects:	Q Science > QK Botany
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	21 Feb 2012 18:08
Last Modified:	21 Feb 2012 18:08
URI:	http://crdd.osdd.net/open/id/eprint/1062

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