Production of dengue 2 envelope domain III in plant using TMV-based vector system.
The envelope protein of dengue virus is the major protein involved in host cell receptor binding for viral entry and induction of immunity. A gene fragment encoding domain III of the dengue 2 envelope protein (D2EIII, amino acids 298-400) was successfully expressed in Nicotinana benthamiana plant using a tobacco mosaic virus (TMV)-based transient expression system. The N-terminal 5' untranslated region-omega sequence located upstream of D2EIII increased protein production in infected plant tissues. The recombinant protein was reactive with anti-D2EIII polyclonal and anti-His tag antibodies. The intramuscular immunization of mice with D2EIII induced the production of the anti-dengue virus antibody. The induced antibody demonstrated neutralizing activity against dengue type 2 virus. The result indicates that the TMV expression system produces the dengue virus antigen in plant, which possesses appropriate antigenicity and immunogenicity.
The International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
Wanida
W
Saejung
Kazuhito
K
Fujiyama
Tomohiko
T
Takasaki
Mikako
M
Ito
Koichi
K
Hori
Prida
P
Malasit
Yuichiro
Y
Watanabe
Ichiro
I
Kurane
Tatsuji
T
Seki
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D007114:Immunization / Q000379:methods; D007273:Injections, Intramuscular; D051379:Mice; D008809:Mice, Inbred C3H; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014027:Tobacco Mosaic Virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; D014765:Viral Vaccines