Cholera toxin B subunit-domain III of dengue virus envelope glycoprotein E fusion protein production in transgenic plants.
Envelope glycoprotein E of the dengue virus, which plays a crucial role in its entry into host cells, has an immunogenic domain III (EIII, amino acids 297-394), which is capable of inducing neutralizing antibodies. However, mice immunized with EIII protein without adjuvant elicited low immune responses. To improve low immune responses, a DNA fragment, consisting of cholera toxin B subunit and EIII gene (CTB-EIII), was constructed and introduced into tobacco plant cells (Nicotiana tabacum L. cv. MD609) by Agrobacterium tumefaciens-mediated transformation methods. The integration and transcription of CTB-EIII fusion gene were confirmed in transgenic plants by genomic DNA PCR amplification and Northern blot analysis, respectively. The results of immunoblot analysis with anti-CTB and anti-dengue virus antibodies showed the expression of the CTB-EIII fusion protein in transgenic plant extracts. Based on the G(M1)-ELISA results, the CTB-EIII protein expressed in plants showed the biological activity for intestinal epithelial cell membrane glycolipid receptor, G(M1)-ganglioside, and its expression level was up to about 0.019% of total soluble protein in transgenic plant leaf tissues. The feasibility of using a plant-produced CTB-EIII fusion protein to generate immunogenicity against domain III will be tested in future animal experiments.
Kim
Tae-Geum
TG
Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea.
Kim
Mi-Young
MY
Yang
Moon-Sik
MS
D002772:Cholera Toxin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D053059:Dengue Vaccines / Q000276:immunology; D003716:Dengue Virus / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D030821:Plants, Genetically Modified / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics; D014026:Nicotiana / Q000235:genetics; D022462:Vaccines, Edible / Q000276:immunology
D053059:Dengue Vaccines; D006023:Glycoproteins; D011994:Recombinant Proteins; D022462:Vaccines, Edible; D002772:Cholera Toxin