茶尺蠖小RNA病毒5'端非编码区的克隆和测序及与哺乳动物小RNA病毒的比较分析

›› 2004, Vol. 47 ›› Issue (5): 573-578.

茶尺蠖小RNA病毒5'端非编码区的克隆和测序及与哺乳动物小RNA病毒的比较分析

王小纯^1,2，张珈敏¹，蒋洪¹，俞海洋¹，谭莉¹，胡远扬^1*

出版日期:2004-10-20 发布日期:2004-10-20

Cloning and sequencing of the 5'untranslation region of Ectropis oblique picornavirus with a comparison to that of mammalian picornaviruses

WANG Xiao-Chun^1，2, ZHANG Jia-Min¹, JIANG Hong¹, YU Hai-Yang¹, TAN-Li¹, HU Yuan-Yang^1*

Online:2004-10-20 Published:2004-10-20
Contact: HU Yuan-Yang

摘要/Abstract

摘要： 用Trizol从纯化的茶尺蠖Ectropis oblique小RNA病毒（EoPV）中提取病毒基因组RNA，逆转录后加poly(dT)，然后进行两步PCR扩增基因组5′端。克隆测序后，对其5′端非编码区的核苷酸序列进行分析，发现具有哺乳动物小RNA病毒的5′端非编码区的一些特征：A/T含量丰富、起始密码子上游AUG和小顺反子多。利用mfold预测了EoPV 5′端非编码区的二级结构，存在4个茎环结构，有哺乳动物内部核糖体进入位点（IRES）的保守区域，即含保守基序GNRA的茎环A和A/C丰富的环B及多聚嘧啶区域。据此推测EoPV基因组翻译采用IRES起始机制。

关键词: 茶尺蠖小RNA病毒, 哺乳动物小RNA病毒, 5′端非编码区, 结构分析

Abstract: After the genomic RNA was extracted from purified Ectropis oblique picornavirus (EoPV) virons with Trizol, cDNA fragments complementary to EoPV RNA were synthesized using reverse transcriptase and added poly (dT) tail. The 5′terminus was amplified by two step PCR. After being cloned and sequenced, the 5′UTR of EoPV was compared with that of mammalian picornaviruses. Like the 5′UTRs of mammalian picornaviruses, the 5′UTR of EoPV was rich in A/T, and had many AUG and small cistrons positioned upstream from the authentic initiation codon. The second structure of 5′UTR of EoPV was predicted with the mfold software. There were 4 stem loops, and conserved motifs of mammalian IRESs, including stemloop A, loop B (A/C rich) and polypyrimidine tract. Based on the characteristics of 5′UTR of EoPV, the genome was suggested to be translated with IRES mechanism.

Key words: Ectropis oblique picornavirus, mammalian picornaviruses, 5'untranslation region, structural analysis

王小纯^1,2，张珈敏¹，蒋洪¹，俞海洋¹，谭莉¹，胡远扬^1*. 茶尺蠖小RNA病毒5'端非编码区的克隆和测序及与哺乳动物小RNA病毒的比较分析[J]. , 2004, 47(5): 573-578.

WANG Xiao-Chun^1，2, ZHANG Jia-Min¹, JIANG Hong¹, YU Hai-Yang¹, TAN-Li¹, HU Yuan-Yang^1*. Cloning and sequencing of the 5'untranslation region of Ectropis oblique picornavirus with a comparison to that of mammalian picornaviruses[J]. , 2004, 47(5): 573-578.

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