Expression and functional validation of heat-labile enterotoxin B (LTB) and cholera toxin B (CTB) subunits in transgenic rice (Oryza sativa)
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- 作者：Ho Seob Soh ; Ha Young Chung ; Hyun Ho Lee ; Hemavathi Ajjappala…
- 关键词：Oryza sativa ; Enterotoxin ; Transgenic plant ; Oral vaccine ; Immunogenicity
- 刊名：SpringerPlus
- 出版年：2015
- 出版时间：December 2015
- 年：2015
- 卷：4
- 期：1
- 全文大小：2769KB
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Ha Young Chung (2)
Hyun Ho Lee (3)
Hemavathi Ajjappala (2)
Kyoungok Jang (3)
Jong-Hwa Park (3)
Joon-Soo Sim (2)
Gee Young Lee (1)
Hyun Ju Lee (1)
Young Hee Han (1)
Jae Wook Lim (1)
Inchan Choi (2)
In Sik Chung (3)
Bum-Soo Hahn (2)
1. Division of Environmental Agricultural Research, Gyeonggido Agricultural Research & Extension Services, Hwaseong, 445-784, South Korea
2. Department of Agricultural Biotechnology, National Academy of Agricultural Science, Nongsaengmyeong-ro 370, Jeonju-si, Jeollabuk-do, 560-550, South Korea
3. Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701, South Korea - 刊物类别：Science, general;
- 刊物主题：Science, general;
- 出版者：Springer International Publishing
- ISSN：2193-1801
文摘
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. Keywords Oryza sativa Enterotoxin Transgenic plant Oral vaccine Immunogenicity