AMBISENSE CODING STRATEGY OF THE RICE STRIPE VIRUS GENOME - INVITRO TRANSLATION STUDIES
AMBISENSE CODING STRATEGY OF THE RICE STRIPE VIRUS GENOME - INVITRO TRANSLATION STUDIES
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DOI：
10.1099/0022-1317-74-6-1125
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发表时间：
1993-06-01
影响因子：
3.8
通讯作者：
ISHIHAMA, A
中科院分区：
文献类型：
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作者：
HAMAMATSU, C;TORIYAMA, S;ISHIHAMA, A
水稻条纹病毒（RSV）的基因组包含4个RNA片段，是水稻条纹病毒（RSV）的典型种。对三个小片段的序列分析表明，它们都具有双义编码策略。为了检验基因组RNA的双义性，我们使用T7 RNA聚合酶在体外合成了携带假定的开放阅读框（ORFs）的RNA，通过转录RNA片段2、3和4的cDNA克隆，并在体外使用两种系统（网状细胞裂解物和小麦胚芽提取物）翻译每个RNA。我们检测了存在于病毒rna （vRNAs）及其互补rna （cRNAs）的5'-近端区域的orf编码的蛋白质。总vRNA在体外翻译中产生由vRNA 5′区orf编码的蛋白。总体结果与RSV rna的预测一致，至少在片段2之前，它们的编码策略是双义的。
Rice stripe virus (RSV), the type species of the tenuivirus group, contains four RNA segments as its genome. Sequence analyses of the three smaller segments indicated that all of them have ambisense coding strategies. To examine the ambisense nature of the genomic RNAs, we synthesized in vitro the RNAs carrying the putative open reading frames (ORFs) by transcribing cDNA clones for RNA segments 2, 3 and 4 in both directions using T7 RNA polymerase and translated each RNA in vitro using two systems: reticulocyte lysates and wheat-germ extracts. We detected the proteins encoded by the ORFs present in the 5'-proximal regions of both viral RNAs (vRNAs) and their complementary RNAs (cRNAs). Translation in vitro of total vRNA generated proteins encoded by the ORFs present in the 5' regions of vRNAs. The overall results are consistent with the prediction that RSV RNAs, at least up to segment 2, are ambisense in their coding strategy.