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莴苣传染性黄化病毒基因组RNA的部分特征分析、外壳蛋白基因的鉴定及其氨基酸序列与其他丝状RNA植物病毒的比较

Partial characterization of the lettuce infectious yellows virus genomic RNAs, identification of the coat protein gene and comparison of its amino acid sequence with those of other filamentous RNA plant viruses.

作者信息

Klaassen V A, Boeshore M, Dolja V V, Falk B W

机构信息

Department of Plant Pathology, University of California, Davis 95616.

出版信息

J Gen Virol. 1994 Jul;75 ( Pt 7):1525-33. doi: 10.1099/0022-1317-75-7-1525.

Abstract

Purified virions of lettuce infectious yellows virus (LIYV), a tentative member of the closterovirus group, contained two RNAs of approximately 8500 and 7300 nucleotides (RNAs 1 and 2 respectively) and a single coat protein species with M(r) of approximately 28,000. LIYV-infected plants contained multiple dsRNAs. The two largest were the correct size for the replicative forms of LIYV virion RNAs 1 and 2. To assess the relationships between LIYV RNAs 1 and 2, cDNAs corresponding to the virion RNAs were cloned. Northern blot hybridization analysis showed no detectable sequence homology between these RNAs. A partial amino acid sequence obtained from purified LIYV coat protein was found to align in the most upstream of four complete open reading frames (ORFs) identified in a LIYV RNA 2 cDNA clone. The identity of this ORF was confirmed as the LIYV coat protein gene by immunological analysis of the gene product expressed in vitro and in Escherichia coli. Computer analysis of the LIYV coat protein amino acid sequence indicated that it belongs to a large family of proteins forming filamentous capsids of RNA plant viruses. The LIYV coat protein appears to be most closely related to the coat proteins of two closteroviruses, beet yellows virus and citrus tristeza virus.

摘要

生菜传染性黄化病毒(LIYV)是长线形病毒属的一个暂定成员,其纯化病毒粒子含有两条分别约为8500和7300个核苷酸的RNA(分别为RNA 1和RNA 2)以及一种单一的外壳蛋白,其相对分子质量约为28,000。感染LIYV的植物含有多个双链RNA。其中两条最大的双链RNA大小与LIYV病毒粒子RNA 1和RNA 2的复制形式相符。为了评估LIYV的RNA 1和RNA 2之间的关系,克隆了与病毒粒子RNA对应的cDNA。Northern杂交分析表明,这些RNA之间没有可检测到的序列同源性。从纯化的LIYV外壳蛋白获得的部分氨基酸序列,与在LIYV RNA 2 cDNA克隆中鉴定出的四个完整开放阅读框(ORF)中最上游的一个排列一致。通过对体外和在大肠杆菌中表达的基因产物进行免疫学分析,确认该ORF的身份为LIYV外壳蛋白基因。对LIYV外壳蛋白氨基酸序列的计算机分析表明,它属于形成RNA植物病毒丝状衣壳的一大类蛋白质。LIYV外壳蛋白似乎与两种长线形病毒,即甜菜黄化病毒和柑橘衰退病毒的外壳蛋白关系最为密切。

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