Risk-managed production of bioactive recombinant proteins using a novel plant virus vector with a helper plant to complement viral systemic movement
Risk-managed production of bioactive recombinant proteins using a novel plant virus vector with a helper plant to complement viral systemic movement
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DOI：
10.1111/j.1467-7652.2010.00529.x
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发表时间：
2011-01-01
影响因子：
13.8
通讯作者：
Matsumura, Takeshi
中科院分区：
文献类型：
--
作者：
Fukuzawa, Noriho;Ishihara, Takeaki;Matsumura, Takeshi
植物病毒载体具有在短时间内以低成本有效生产重组蛋白的潜力。虽然重组蛋白也可以通过转基因植物生产，但当需要紧急扩大生产规模时，植物病毒载体（如果可用）可能更方便。然而，由于存在逃逸到环境中的风险，很难在开阔的田野中使用病毒载体。在这项研究中，我们构建了一个新的病毒载体系统，使用运动缺陷型黄瓜花叶病毒（CMV）载体，这是理论上本地化的接种细胞，但感染系统只有借助转基因辅助植物，补充病毒运动，减少病毒增殖的风险。有趣的是，用载体系统感染的辅助植物对野生型CMV的攻击接种产生了强的交叉保护。使用属于两个离散的CMV组（亚组I和II）的CMV株系，我们还改进了系统，以防止载体和辅助植物中的转基因转录本之间的重组。我们在这里展示了表达的抗二恶英单链可变区片段（DxscFv）和白细胞介素-1受体拮抗剂（IL-1-Ra）在烟草本塞姆亚纳通过这种病毒载体限制系统，这是适用于许多有用的高品质的重组蛋白。
A plant viral vector has the potential to efficiently produce recombinant proteins at a low cost in a short period. Although recombinant proteins can be also produced by transgenic plants, a plant viral vector, if available, may be more convenient when urgent scale-up in production is needed. However, it is difficult to use a viral vector in open fields because of the risk of escape to the environment. In this study, we constructed a novel viral vector system using a movement-defective Cucumber mosaic virus (CMV) vector, which is theoretically localized in the inoculated cells but infects systemically only with the aid of the transgenic helper plant that complements viral movement, diminishing the risk of viral proliferation. Interestingly, the helper plant systemically infected with the vector gave strong cross-protection against challenge inoculation with wild-type CMVs. Using CMV strains belonging to two discrete CMV groups (subgroups I and II), we also improved the system to prevent recombination between the vector and the transgene transcript in the helper plant. We here demonstrate the expression of an anti-dioxin single chain variable fragment (DxscFv) and interleukin-1 receptor antagonist (IL1-Ra) in Nicotiana benthamiana by this viral vector confinement system, which is applicable for many useful high-quality recombinant proteins.