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)

Sharma, Manoj Kumar ; Singh, Nirmal Kumar ; Jani, Dewal ; Sisodia, Rama ; Thungapathra, M. ; Gautam, J. K. ; Meena, L. S. ; Singh, Yogendra ; Ghosh, Amit ; Tyagi, Akhilesh Kumar ; et., al (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. 307-318. ISSN 0721-7714

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Official URL: http://www.springerlink.com/content/706356232827g3...

Related URL: http://dx.doi.org/10.1007/s00299-007-0464-y

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 GM1-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
Source:Copyright of this article belongs to Springer-Verlag.
Keywords:Cholera; Fusion Protein; TCPA; Tomato; Transgenic
ID Code:11085
Deposited On:09 Nov 2010 04:08
Last Modified:10 Dec 2011 08:53

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