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.
Transgenic Res. 2010 Oct;19(5):889-95. doi: 10.1007/s11248-009-9358-9. Epub 2010 Jan 14.
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.
HIV-1 Tat 在病毒复制中起着重要作用，是艾滋病发展的关键，使其成为诱导体液和细胞免疫的理想疫苗靶点。由于成本更低，植物性抗原生产似乎是疫苗生产的理想选择。在这项研究中，我们创建了一个植物优化的 tat 和突变体（Cys30Ala/Lys41Ala）tat（mtat）基因，并将每个基因连接到带有终止密码子和 GusA 基因的 pBI121 表达载体上，GusA 基因位于其下游。载体构建体被轰击到番茄叶片愈伤组织中，并允许其发育。因此，我们生成了优先表达 Tat-GUS 融合蛋白而不是 Tat 蛋白的重组番茄植物。此外，用 tat 或 mtat 基因轰击的植物没有表现出表型差异，并且每毫克可溶性植物蛋白产生 2-4 微克 Tat-GUS 融合蛋白。此外，发现皮内接种到小鼠体内的番茄提取物可诱导体液免疫，最重要的是，还可诱导细胞免疫。
