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构建土壤传播小麦花叶病毒RNA1和RNA2的全长cDNA克隆,体外转录产生感染性RNA:病毒粒子形成及全身感染,衣壳蛋白的N端和C端延伸区不表达。

Construction of full-length cDNA clones to Soil-borne wheat mosaic virus RNA1 and RNA2, from which infectious RNAs are transcribed In vitro: virion formation and systemic infection without expression of the N-terminal and C-terminal extensions to the capsid protein.

作者信息

Yamamiya A, Shirako Y

机构信息

Asian Center for Bioresources and Environmental Sciences (ANESC), Graduate School of Agricultural Life Sciences, Tokyo, Japan.

出版信息

Virology. 2000 Nov 10;277(1):66-75. doi: 10.1006/viro.2000.0587.

Abstract

The 19-kDa capsid protein (CP) of Soil-borne wheat mosaic furovirus (SBWMV) is encoded in the 5'-terminal region of RNA2. In addition to CP, two CP-related proteins are translated from SBWMV RNA2: (1) a 24-kDa protein (N-CP) with an N-terminal 40-amino-acid extension initiated at an upstream in-frame CUG codon; and (2) an 83-kDa protein (CP-RT) with an about 580-amino-acid, C-terminal extension by partial translational readthrough at the UGA termination codon at the end of the CP gene. We examined requirements for N-CP and CP-RT on virion formation and systemic infection in wheat plants using full-length cDNA clones, from which infectious RNA can be transcribed in vitro. RNA2 mutants, which could not synthesize N-CP, CP-RT, or either infected wheat plants systemically in combination with the wild-type RNA1 transcripts, produced rod-shaped virus particles in uninoculated upper leaves. Original mutations which abolished translation of N-CP and CP-RT were confirmed on RNA2 extracted from purified virus from the upper leaves by nucleotide sequence analysis. These results indicate that neither N-terminal nor C-terminal extensions to the CP are required for virion formation and systemic infection of SBWMV in wheat plants.

摘要

土传小麦花叶真菌传杆状病毒(SBWMV)的19-kDa衣壳蛋白(CP)由RNA2的5'-末端区域编码。除CP外,从SBWMV RNA2还翻译出两种与CP相关的蛋白:(1)一种24-kDa蛋白(N-CP),其N末端有40个氨基酸的延伸,起始于上游的框内CUG密码子;(2)一种83-kDa蛋白(CP-RT),其C末端通过在CP基因末端的UGA终止密码子处的部分翻译通读而有大约580个氨基酸的延伸。我们使用全长cDNA克隆研究了N-CP和CP-RT在小麦植株病毒粒子形成和系统感染中的需求,从该克隆可在体外转录出感染性RNA。不能合成N-CP、CP-RT或两者的RNA2突变体,与野生型RNA1转录本一起可使小麦植株系统感染,在未接种的上部叶片中产生杆状病毒粒子。通过核苷酸序列分析在从上部叶片纯化病毒中提取的RNA2上证实了消除N-CP和CP-RT翻译的原始突变。这些结果表明,对于SBWMV在小麦植株中的病毒粒子形成和系统感染,CP的N末端和C末端延伸均非必需。

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