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A defective RNA associated with bamboo mosaic virus and the possible common mechanisms for RNA recombination in potexviruses.

机译:与竹花叶病毒有关的有缺陷的RNA,以及在potexviruses中RNA重组的可能常见机制。

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摘要

A naturally occurring 1.1 kb RNA was isolated from purified virions of bamboo mosaic potexvirus isolate S (BaMV-S). This RNA is a defective RNA (D RNA) derived from a single internal deletion of the BaMV genome. A cDNA clone representing the complete nucleotide sequence of the BaMV-S D RNA was generated and its nucleotide sequence was determined. The BaMV D cDNA is 1015 nts in length [excluding the poly(A) tail] and consists of two regions corresponding to 867 nts of the 5' terminus and 148 nts of the 3' terminus of the BaMV genomic RNA. BaMV D cDNA contains a single open reading frame (ORF) encoding a putative 29.7 kDa protein comprised of a fusion of the first 258 amino acids of BaMV ORF 1 and the last 2 amino acids of coat protein. The coding capacity of D RNA was verified by in vitro translation of native BaMV-S D RNA and of 1.1 kb RNA transcribed in vitro from the full-length D cDNA. BaMV D RNA can be reproducibly generated by serial passages of BaMV-S in Nicotiana benthamiana and is the first D RNA in the potexvirus group shown to be generated de novo. Alignments of sequences surrounding the 5' and 3' junction borders of reported potexvirus D RNAs reveal a 65.2-84.6% sequence identity, suggesting that common mechanisms for viral RNA recombination are involved in the generation of potexvirus D RNAs.
机译:从竹花叶病毒分离株S(BaMV-S)的纯化病毒体中分离出天然存在的1.1 kb RNA。此RNA是源自BaMV基因组的单个内部缺失的缺陷RNA(D RNA)。产生了代表BaMV-S D RNA完整核苷酸序列的cDNA克隆,并确定了其核苷酸序列。 BaMV D cDNA的长度为1015 nts(不包括poly(A)尾巴),由两个区域组成,分别对应于BaMV基因组RNA的5'端的867 nt和3'端的148 nt。 BaMV D cDNA包含一个单一的开放阅读框(ORF),编码一个假定的29.7 kDa蛋白,该蛋白由BaMV ORF 1的前258个氨基酸与外壳蛋白的后2个氨基酸融合而成。 D RNA的编码能力是通过体外翻译天然BaMV-S D RNA和从全长D cDNA体外转录的1.1 kb RNA来验证的。 BaMV D RNA可以通过本氏烟草中BaMV-S的连续传代而可再生地产生,并且是potexvirus组中从头产生的第一个D RNA。报道的potexvirus D RNA的5'和3'交界处的序列比对显示出65.2-84.6%的序列同一性,表明病毒RNA重组的共同机制与potexvirus D RNA的产生有关。

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