一个RNA沉默抑制器的识别被一个印度类环点病毒编码
Aniket Angira1, V K Baranwal2, Aashish Ranjan3
1Amity Institute of Virology & Immunology, Amity University Uttar Pradesh, Noida, Uttar Pradesh 201313 India.
概括
印度类环点病毒 (ICRSV) ORF5蛋白抑制宿主RNAi沉默. 这种蛋白质作为RNA沉默抑制剂 (RSS),保护病毒,并可能有助于类植物的疾病发展.
科学领域:
- 植物病毒学 植物病毒学
- 分子生物学分子生物学
- 生物化学 生化学
背景情况:
- 植物病毒利用RNA沉默抑制剂 (RSS) 逃避宿主抗病毒防御,这是病毒生存的关键机制.
- 印度环点病毒 (ICRSV) 在金诺生产中造成重大经济损失,但其RSS蛋白仍然未被确定.
- 了解ICRSV的RSS对于开发有效的疾病管理策略至关重要.
研究的目的:
- 为了识别特定的开放阅读框架 (ORF) 编码印度类环状点病毒 (ICRSV) 的RNA沉默抑制剂 (RSS) 蛋白质.
- 功能性地描述已识别的蛋白质抑制宿主诱导的RNA干扰 (RNAi) 的能力.
主要方法:
- 将ICRSV ORFs (2, 3, 4, 5, 和 6) 克隆成植物表达载体 (pCAMBIA1302-35s-GFP).
- 构造物的农业透到 *Nicotiana benthamiana * 叶子中,以评估绿色光蛋白 (GFP) 沉默抑制.
- 使用β-glucuronidase (GUS) 报告器和分子分析 (半定量PCR,qRT-PCR) 的基因分析以确认基因表达和沉默抑制.
主要成果:
- 使用ORF5结构的农业透导致持续的GFP光,表明抑制RNAi介导的沉默,类似于已知的RSS (P19).
- 用ORF5-GUS进行的β-葡萄糖酶 (GUS) 测定也表明了沉默抑制,由diX-色沉物形成证明.
- 分子分析证实ORF5表达叶子中GFP和GUS记者基因的表达水平较高,证实了其沉默抑制活性.
结论:
- 由ICRSV的ORF5编码的蛋白质具有显著的RNA沉默抑制剂 (RSS) 活性.
- ICRSV ORF5蛋白有效抵消宿主的RNAi防御机制,有助于病毒的持久性.
- 这一发现确定了与ICRSV病原发生有关的关键病毒因素,并为未来的控制策略提供了目标.
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