Yeh T Y, Lin B Y, Chang Y C, Hsu Y H, Lin N S
Institute of Botany, Academia Sinica, Taipei, Taiwan, Republic of China.
Virus Genes. 1999;18(2):121-8. doi: 10.1023/a:1008008400653.
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.
从竹花叶病毒分离株S(BaMV-S)的纯化病毒粒子中分离出一种天然存在的1.1 kb RNA。这种RNA是一种缺陷RNA(D RNA),源自BaMV基因组的单个内部缺失。生成了一个代表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长度为1015个核苷酸[不包括聚(A)尾],由两个区域组成,分别对应于BaMV基因组RNA 5'末端的867个核苷酸和3'末端的148个核苷酸。BaMV D cDNA包含一个单一的开放阅读框(ORF),编码一种推定的29.7 kDa蛋白质,该蛋白质由BaMV ORF 1的前258个氨基酸和衣壳蛋白的最后2个氨基酸融合而成。通过对天然BaMV-S D RNA和从全长D cDNA体外转录的1.1 kb RNA进行体外翻译,验证了D RNA的编码能力。BaMV D RNA可以通过BaMV-S在本氏烟草中连续传代而可重复产生,并且是在马铃薯X病毒组中显示可从头产生的第一个D RNA。已报道的马铃薯X病毒D RNA的5'和3'连接边界周围序列的比对显示序列同一性为65.2-84.6%,这表明病毒RNA重组的共同机制参与了马铃薯X病毒D RNA的产生。