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Molecular Analysis of Replication and Packaging Mechanism of Satellite Tobacco Mosaic Virus.

机译:卫星烟草花叶病毒复制和包装机制的分子分析。

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摘要

Satellite tobacco mosaic virus (STMV) as the name implies is the satellite virus of the well characterized helper Tobacco mosaic virus (TMV). This satellite virus is 18nm in diameter having a T1 symmetry encapsidating a positive sense genomic RNA of 1058nt in size whose 3' 150nt shares 65% homology with that of its Helper virus. Extensive Structural studies on STMV suggested that its coat protein interacts extensively with the stem-loops of the packaged genomic RNA and hence would be unstable to form virions without interacting with its genomic RNA internally. However invivo experiments to prove the same were not performed. Host range effects and the role of the positively charged amino acids at the N-terminal of its capsid protein on replication and packaging were not been studied yet on this Satellite virus (or in any other satellite virus). Chapter 1 of this dissertation describes T-DNA based agrotransformants of STMV to synthesize its genomic RNA in either (+) or (-) orientation when expressed ectopically in Nicotiana benthamiana plants. The biological activity of ectopically expressed agrotransformants of STMV (+) or (-) transcripts was confirmed when co-expressed with TMV. Furthermore, progeny virions of STMV recovered from agroinfiltrations were indistinguishable from those recovered from mechanically inoculated plants. However, STMV virions assembled in N. benthamiana, but not in tobacco, efficiently packaged a variant form of the genomic RNA. Chapter 2 describes Individual substitution of the positively charged amino acids at the N-terminal region of its Coat protein to a neutrally charged amino acid Alanine affecting dramatically the replication and packaging of these mutant viruses. Chapter 3 describes Agrobacterium mediated transient expression of the three genomic RNA of Tomato Aspermy virus in N.benthamiana and by utilizing this system we demonstrate that expressed genomic RNA were biologically active and also induced symptoms in N.benthamiana.
机译:顾名思义,卫星烟草花叶病毒(STMV)是特征明确的辅助烟草花叶病毒(TMV)的卫星病毒。该卫星病毒直径为18nm,具有T1对称性,囊括了大小为1058nt的正向基因组RNA,其3'150nt与它的辅助病毒具有65%的同源性。对STMV进行的广泛结构研究表明,其外壳蛋白与包装的基因组RNA的茎环广泛相互作用,因此在不与内部基因组RNA相互作用的情况下,其形成病毒体的过程将不稳定。但是,并未进行证明相同的体内实验。尚未在该卫星病毒(或任何其他卫星病毒)中研究宿主范围效应及其衣壳蛋白N末端带正电荷的氨基酸对复制和包装的作用。本论文的第1章描述了在烟草本生植物中异位表达时,基于T-DNA的STMV农杆菌转化子以(+)或(-)方向合成其基因组RNA。当与TMV共表达时,证实了异位表达的STMV(+)或(-)转录本的农转化子的生物活性。此外,从农杆菌渗入中回收的STMV的子代病毒体与从机械接种的植物中回收的那些并没有区别。然而,在本氏烟草中组装而不是在烟草中组装的STMV病毒体有效地包装了基因组RNA的变体形式。第2章介绍了将其外壳蛋白N端区域的带正电荷的氨基酸分别替换为中性带电荷的氨基酸丙氨酸的方法,这些氨基酸极大地影响了这些突变病毒的复制和包装。第3章描述了农杆菌介导的番茄曲霉病毒三个基因组RNA在本氏烟草中的瞬时表达,通过利用该系统,我们证明了所表达的基因组RNA具有生物学活性,并在本氏烟草中引起症状。

著录项

  • 作者

    Sivanandam, Venkatesh.;

  • 作者单位

    University of California, Riverside.;

  • 授予单位 University of California, Riverside.;
  • 学科 Biology Virology.;Agriculture Plant Pathology.
  • 学位 Ph.D.
  • 年度 2014
  • 页码 113 p.
  • 总页数 113
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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