Preferential expression and immunogenicity of HIV-1 Tat fusion protein expressed in tomato plant
Preferential expression and immunogenicity of HIV-1 Tat fusion protein expressed in tomato plant
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DOI：
10.1007/s11248-009-9358-9
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发表时间：
2010-10-01
影响因子：
3
通讯作者：
Okamoto, Takashi
中科院分区：
文献类型：
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作者：
Cueno, Marni E.;Hibi, Yurina;Okamoto, Takashi
HIV-1达特在病毒复制中起主要作用，并且对于AIDS的发展是必需的，使得其成为理想的疫苗靶标，只要诱导体液和细胞免疫应答。基于植物的抗原生产，由于其成本较低，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达特和突变体（Cys 30 Ala/Lys 41 Ala）达特（mtat）基因，并连接到一个pBI 121表达载体与终止密码子和gusA基因定位在紧下游。将载体构建体轰击到番茄叶愈伤组织中并使其发育。因此，我们产生了重组番茄植物优先表达Tat-GUS融合蛋白超过Tat-唯一的蛋白。此外，用达特或mtat基因轰击的植物没有表现出表型差异，每毫克可溶性植物蛋白产生2-4 μ g Tat-GUS融合蛋白。此外，番茄提取物皮内接种到小鼠被发现诱导体液免疫，最重要的是，细胞免疫。
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 mu g 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.