title: Expression and functional validation of heat-labile enterotoxin B (LTB) and cholera toxin B (CTB) subunits in transgenic rice (Oryza sativa).
abstract: 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 GM1-ganglioside binding enzyme-linked immunosorbent assays (GM1-ELISA). Real-time PCR of three generations (T3, T4, and T5) in homozygous lines (LCI-11) showed single copies of LTB, CTB, bar and Tnos. 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.
authors: Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784 South Korea.
authors: Ho Seob
authors: HS
authors: Soh
authors: Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do 560-550 South Korea.
authors: Ha Young
authors: HY
authors: Chung
authors: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701 South Korea.
authors: Hyun Ho
authors: HH
authors: Lee
authors: Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do 560-550 South Korea.
authors: Hemavathi
authors: H
authors: Ajjappala
authors: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701 South Korea.
authors: Kyoungok
authors: K
authors: Jang
authors: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701 South Korea.
authors: Jong-Hwa
authors: JH
authors: Park
authors: Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do 560-550 South Korea.
authors: Joon-Soo
authors: JS
authors: Sim
authors: Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784 South Korea.
authors: Gee Young
authors: GY
authors: Lee
authors: Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784 South Korea.
authors: Hyun Ju
authors: HJ
authors: Lee
authors: Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784 South Korea.
authors: Young Hee
authors: YH
authors: Han
authors: Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784 South Korea.
authors: Jae Wook
authors: JW
authors: Lim
authors: Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do 560-550 South Korea.
authors: Inchan
authors: I
authors: Choi
authors: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701 South Korea.
authors: In Sik
authors: IS
authors: Chung
authors: Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do 560-550 South Korea.
authors: Bum-Soo
authors: BS
authors: Hahn
keywords: Enterotoxin; Immunogenicity; Oral vaccine; Oryza sativa; Transgenic plant