Complete nucleotide sequence and genome organization of hibiscus chlorotic ringspot virus, a new member of the genus Carmovirus:: Evidence for the presence and expression of two novel open reading frames
Complete nucleotide sequence and genome organization of hibiscus chlorotic ringspot virus, a new member of the genus Carmovirus:: Evidence for the presence and expression of two novel open reading frames
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DOI：
10.1128/jvi.74.7.3149-3155.2000
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发表时间：
2000-04-01
影响因子：
5.4
通讯作者：
Wong, SM
中科院分区：
文献类型：
--
作者：
Huang, M;Koh, DCY;Wong, SM
测定了芙蓉绿环斑病毒（HCRSV）的全核苷酸序列。基因组RNA （gRNA）全长3,911个核苷酸，可能编码28 （p28）、23 （p23）、81 （p81）、8 （p8）、9 （p9）、38 （p38）和25 (p25) kDa的7种病毒蛋白。除了编码p23和p25的两个独特的开放阅读框（orf）外，orf编码的蛋白质与carmovirus具有高度的氨基酸相似性，除了gRNA外，还鉴定出两个3'共端亚基因组RNA （sgRNA）种。在T7启动子的控制下，构建了gRNA和sgRNA的全长cDNA克隆。来自全长基因组cDNA克隆的带帽转录本和未带帽转录本均具有传染性。体外翻译和诱变实验证实，除了编码p8的ORF外，所有预测的ORF都是可翻译的，并且两个新的ORF（编码p23和p25的ORF）可能在病毒感染周期的功能上不可或缺。基于病毒粒子形态和基因组结构，我们建议将HCRSV归为tombusvirus科Carmovirus属的新成员。
The complete nucleotide sequence of hibiscus chlorotic ringspot virus (HCRSV) was determined. The genomic RNA (gRNA) is 3,911 nucleotides long and has the potential to encode seven viral proteins in the order of 28 (p28), 23 (p23), 81 (p81), 8 (p8), 9 (p9), 38 (p38), and 25 (p25) kDa, Excluding two unique open reading frames (ORFs) encoding p23 and p25, the ORFs encode proteins with high amino acid similarity to those of carmoviruses, In addition to gRNA, two 3'-coterminated subgenomic RNA (sgRNA) species were identified. Full-length cDNA clones derived from gRNA and sgRNA were constructed under the control of a T7 promoter. Both capped and uncapped transcripts derived from the full-length genomic cDNA clone were infectious. In vitro translation and mutagenesis assays confirmed that all the predicted ORFs except the ORF encoding p8 are translatable, and the two novel ORFs (those encoding p23 and p25) may be functionally indispensable for the viral infection cycle. Based on virion morphology and genome organization, we propose that HCRSV be classified as a new member of the genus Carmovirus in family Tombusviridae.