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细菌表达的针对烟草花叶病毒或马铃薯 Y 病毒基因组热点序列的双链 RNA 具有不同的保护烟草免受病毒感染的能力。

Bacterially expressed double-stranded RNAs against hot-spot sequences of tobacco mosaic virus or potato virus Y genome have different ability to protect tobacco from viral infection.

机构信息

Shandong Agricultural University, Tai'an, China.

出版信息

Appl Biochem Biotechnol. 2010 Nov;162(7):1901-14. doi: 10.1007/s12010-010-8968-2. Epub 2010 May 2.

Abstract

Posttranscriptional gene silencing, also known as RNA interference, involves degradation of homologous mRNA sequences in organisms. In plants, posttranscriptional gene silencing is part of a defense mechanism against virus infection, and double-stranded RNA is the pivotal factor that induces gene silencing. In this paper, we got seven hairpin RNAs (hpRNAs) constructs against different hot-spot sequences of Tobacco mosaic virus (TMV) or Potato virus Y (PVY) genome. After expression in Escherichia coli HT115, we extracted the seven hpRNAs for the test in tobacco against TMV or PVY infection. The data suggest that different hpRNAs against different hot-spot sequences of TMV or PVY genome had different ability to protect tobacco plants from viral infection. The resistance to TMV conferred by the hpRNA against the TMV movement protein was stronger than other TMV hpRNAs; the resistance to PVY conferred by the hpRNA against the PVY nuclear inclusion b was better than that induced by any other PVY hpRNAs. Northern blotting of siRNA showed that the resistance was indeed an RNA-mediated virus resistance.

摘要

转录后基因沉默,也被称为 RNA 干扰,涉及到生物体中同源 mRNA 序列的降解。在植物中,转录后基因沉默是植物抵抗病毒感染的防御机制的一部分,双链 RNA 是诱导基因沉默的关键因素。在本文中,我们构建了七个针对烟草花叶病毒(TMV)或马铃薯 Y 病毒(PVY)基因组不同热点序列的发夹 RNA(hpRNA)。在大肠杆菌 HT115 中表达后,我们提取了这七个 hpRNA,用于烟草对 TMV 或 PVY 感染的检测。结果表明,针对 TMV 或 PVY 基因组不同热点序列的不同 hpRNA 对烟草植株的抗病毒侵染能力不同。针对 TMV 运动蛋白的 hpRNA 赋予烟草对 TMV 的抗性强于其他 TMV hpRNA;针对 PVY 核包含体 b 的 hpRNA 赋予烟草对 PVY 的抗性优于任何其他 PVY hpRNA。siRNA 的 Northern 印迹表明,这种抗性确实是一种 RNA 介导的病毒抗性。

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