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We expressed the heat-labile enterotoxin B (LTB) subunit from enterotoxigenic  Escherichia coli  and the cholera toxin B (CTB) subunit from  Vibrio cholerae  under the control of the rice ( Oryza sativa ) globulin ( Glb ) promoter. Binding of recombinant LTB and CTB proteins was confirmed based on G M1 -ganglioside binding enzyme-linked immunosorbent assays (G M1 -ELISA). Real-time PCR of three generations (T 3 , T 4 , and T 5 ) in homozygous lines (LCI-11) showed single copies of  LTB, CTB, bar  and T nos . LTB and CTB proteins in rice transgenic lines were detected by Western blot analysis. Immunogenicity trials of rice-derived CTB and LTB antigens were evaluated through oral and intraperitoneal administration in mice, respectively. The results revealed that LTB- and CTB-specific IgG levels were enhanced in the sera of intraperitoneally immunized mice. Similarly, the toxin-neutralizing activity of CTB and LTB in serum of orally immunized mice was associated with elevated levels of both IgG and IgA. The results of the present study suggest that the combined expression of CTB and LTB proteins can be utilized to produce vaccines against enterotoxigenic strains of  Escherichia coli  and  Vibrio cholera , for the prevention of diarrhea.

Expression and functional validation of heat-labile enterotoxin B (LTB) and cholera toxin B (CTB) subunits in transgenic rice (Oryza sativa)