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工程稳定传染性cDNA构造的光标记的番茄化病毒的工程

Sun-Jung Kwon¹, Ye-Ji Lee², Young-Eun Cho¹

¹Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Republic of Korea.

|February 16, 2024

概括

研究人员制造了一种绿色光蛋白 (GFP) 标记的番茄化病毒 (ToCV) 来追踪植物中的感染. 该工具可以实时可视化ToCV的运动及其与其他病毒的相互作用.

科学领域:

植物病理学植物病理学
分子病毒学分子病毒学.
生物技术是生物技术.

背景情况:

番茄化病毒 (ToCV) 由于黄叶疾病,导致番茄植物的产量大幅下降.
实时监测病毒感染的有效工具对于了解疾病动态和制定控制策略至关重要.

研究的目的:

开发一种用绿色光蛋白 (GFP) 标签设计的重组ToCV,用于可视化活植物中的病毒感染和运动.
评估复合病毒在混合感染期间研究病毒相互作用的实用性.

主要方法:

设计了ToCV RNA1的全长cDNA结构,并插入了植物内核,以克服细菌转化期间的不稳定性.
将GFP标签与ToCV RNA2的cDNA结构融合在一起.
用重组ToCV-GFP注射尼科蒂亚娜本塔米亚纳植物以确认系统感染并观察GFP表达模式.

主要成果:

成功生成了一个稳定的,传染性重组ToCV-GFP.
在N. benthamiana观察到系统性感染,GFP光在主要静脉沿线局部化,表明病毒运动.
证明了ToCV-GFP在混合感染中揭示ToCV与其他番茄病毒之间的对抗性相互作用的能力.

结论:

开发的ToCV-GFP是实时视觉追踪虫病毒感染和植物运动的宝贵工具.
这种重组病毒有助于在体内研究复杂的病毒-植物和病毒-病毒相互作用.

关键词:

克里尼病毒 (crinivirus) 是一种病毒.在GFP-Tagging中使用.传染性cDNA克隆病毒在Intron 里面.这是一个 ToCVV ToCV.

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