Expression of accessory colonization factor subunit A (ACFA) of Vibrio cholerae and ACFA fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum).
In earlier study from our group, cholera toxin B subunit had been expressed in tomato for developing a plant-based vaccine against cholera. In the present investigation, gene for accessory colonization factor (acf) subunit A, earlier reported to be essential for efficient colonization in the intestine, has been expressed in Escherichia coli as well as tomato plants. Gene encoding for a chimeric protein having a fusion of cholera toxin B subunit and accessory colonization factor A was also expressed in tomato to generate more potent combinatorial antigen. CaMV35S promoter with a duplicated enhancer sequence was used for expression of these genes in tomato. Integration of transgenes into tomato genome was confirmed by PCR and Southern hybridization. Expression of the genes was confirmed at transcript and protein levels. Accessory colonization factor A and cholera toxin B subunit fused to this protein accumulated up to 0.25% and 0.08% of total soluble protein, respectively, in the fruits of transgenic plants. Whereas protein purified from E. coli, in combination with cholera toxin B subunit can be used for development of conventional subunit vaccine, tomato fruits expressing these proteins can be used together with tomato plants expressing cholera toxin B subunit for development of oral vaccine against cholera.
Sharma
Manoj Kumar
MK
Department of Plant Molecular biology, University of Delhi South Campus, New Delhi 110021, India.
Jani
Dewal
D
Thungapathra
M
M
Gautam
J K
JK
Meena
L S
LS
Singh
Yogendra
Y
Ghosh
Amit
A
Tyagi
Akhilesh Kumar
AK
Sharma
Arun Kumar
AK
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002772:Cholera Toxin / Q000235:genetics* / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D018551:Solanum lycopersicum / Q000235:genetics* / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection / Q000379:methods
C093353:AcfA protein, Vibrio cholerae; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D002772:Cholera Toxin