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在表达黄瓜花叶病毒3a基因的转基因烟草中病毒运动的互补作用。

Complementation of virus movement in transgenic tobacco expressing the cucumber mosaic virus 3a gene.

作者信息

Kaplan I B, Shintaku M H, Li Q, Zhang L, Marsh L E, Palukaitis P

机构信息

Department of Plant Pathology, Cornell University, Ithaca, New York 14853, USA.

出版信息

Virology. 1995 May 10;209(1):188-99. doi: 10.1006/viro.1995.1242.

Abstract

Tobacco plants were transformed with the cucumber mosaic cucumovirus (CMV) 3a gene and the in planta-expressed 3a protein was detected immunologically. The 3a protein was predominantly localized in a subcellular fraction corresponding to the cytosol. Two frameshift and four deletion mutants were created within the 3a open reading frame of CMV RNA 3. Five of these mutants, containing an N-terminal, large central, or C-terminal 70-amino-acid deletion could not infect nontransformed tobacco plants, but could infect the 3a transgenic tobacco plants, and generally accumulated to wild-type levels. The sixth mutant, lacking the C-terminal 43 amino acids of the 3a protein, was able to infect nontransformed tobacco plants. A delay in accumulation of viral RNA in both the inoculated and the systemically infected leaves was demonstrated for one of the mutants. Thus, the CMV 3a protein is a virus movement protein, the functions of which can be complemented in a transgenic plant. The CMV 3a transgenic plants were able to complement the long-distance movement of a pseudorecombinant cucumovirus defective for this function in tobacco, as well as the cell-to-cell, but not the long-distance, movement of two other related viruses. However, these transgenic plants were unable to complement the long-distance movement of viruses from several other taxonomic groups that could move cell to cell but not long distance in tobacco.

摘要

用黄瓜花叶黄瓜病毒(CMV)3a基因转化烟草植株,并通过免疫学方法检测在植物中表达的3a蛋白。3a蛋白主要定位于对应于细胞质的亚细胞组分中。在CMV RNA 3的3a开放阅读框内创建了两个移码突变体和四个缺失突变体。这些突变体中的五个,包含N端、大的中央或C端70个氨基酸的缺失,不能感染未转化的烟草植株,但能感染3a转基因烟草植株,并且通常积累到野生型水平。第六个突变体,缺少3a蛋白的C端43个氨基酸,能够感染未转化的烟草植株。其中一个突变体在接种叶和系统感染叶中病毒RNA的积累均出现延迟。因此,CMV 3a蛋白是一种病毒运动蛋白,其功能可在转基因植物中得到互补。CMV 3a转基因植物能够互补在烟草中缺乏此功能的假重组黄瓜病毒的长距离运动,以及另外两种相关病毒的细胞间运动,但不能互补长距离运动。然而,这些转基因植物无法互补来自其他几个分类群的病毒的长距离运动,这些病毒在烟草中能够进行细胞间运动但不能进行长距离运动。

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