Preferential expression and immunogenicity of HIV-1 Tat fusion protein expressed in tomato plant.
HIV-1 Tat plays a major role in viral replication and is essential for AIDS development making it an ideal vaccine target providing that both humoral and cellular immune responses are induced. Plant-based antigen production, due to its cheaper cost, appears ideal for vaccine production. In this study, we created a plant-optimized tat and mutant (Cys30Ala/Lys41Ala) tat (mtat) gene and ligated each into a pBI121 expression vector with a stop codon and a gusA gene positioned immediately downstream. The vector construct was bombarded into tomato leaf calli and allowed to develop. We thus generated recombinant tomato plants preferentially expressing a Tat-GUS fusion protein over a Tat-only protein. In addition, plants bombarded with either tat or mtat genes showed no phenotypic difference and produced 2-4 microg Tat-GUS fusion protein per milligram soluble plant protein. Furthermore, tomato extracts intradermally inoculated into mice were found to induce a humoral and, most importantly, cellular immunity.
Cueno
Marni E
ME
Department of Molecular and Cellular Biology¸ Molecular and Cellular Biology Laboratory, Graduate School of Medical Sciences, Nagoya City University, 1 Kawasumi, Misuho-cho, Mizuho-ku, Nagoya, Aichi, 467-8601, Japan.
Hibi
Yurina
Y
Karamatsu
Katsuo
K
Yasutomi
Yasuhiro
Y
Imai
Kenichi
K
Laurena
Antonio C
AC
Okamoto
Takashi
T
D016915:AIDS Vaccines / Q000276:immunology*; D000818:Animals; D001483:Base Sequence; D017930:Genes, Reporter; D015756:Genes, tat; D005822:Genetic Vectors / Q000235:genetics; D015483:HIV Antibodies / Q000096:biosynthesis; D015497:HIV-1 / Q000235:genetics*; D007074:Immunoglobulin G / Q000096:biosynthesis; D018551:Solanum lycopersicum / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010641:Phenotype; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D017354:Point Mutation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D016915:AIDS Vaccines; D015483:HIV Antibodies; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D054322:tat Gene Products, Human Immunodeficiency Virus