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园艺学报 ›› 2005, Vol. 32 ›› Issue (5): 881-884.

• 研究报告 • 上一篇    下一篇

萝卜中一未知dsRNA全长cDNA克隆及其序列

刘 莉1;陈集双2;喻 珊3;王 冲3
  

  1. (1 湖州师范学院生命科学学院, 湖州313000; 2 浙江理工大学生物工程研究所, 杭州310018; 3 浙江大学生命科学学院, 杭州310029)
  • 收稿日期:2005-02-15 修回日期:2005-05-15 出版日期:2005-10-25 发布日期:2005-10-25

Full-length cDNA Clone and Sequences Analysis for Unknown Double-stranded RNA Isolated from Radish

Liu Li1;Chen Jishuang2;Yu Shan3;Wang Chong3   

  1. (1 Faculty of Life Sciences, Huzhou Teachers College, Huzhou 313000, China; 2 Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou 310018, China; 3College of Life Sciences, Zhejiang University, Hangzhou 310029, China)
  • Received:2005-02-15 Revised:2005-05-15 Online:2005-10-25 Published:2005-10-25

摘要: 采用双链RNA ( double-stranded RNA, dsRNA) 分析法, 发现杭州市近郊的部分萝卜植株及
其子代含有约118 kbp的dsRNA; 在聚丙烯酰胺凝胶上可分离为大小接近的dsRNA1和dsRNA2。以dsRNA1为模板, 采用单引物扩增法获得其全长cDNA; 克隆测序确认为1 866 nt; 预测其最大开放阅读框ORF(Open Reading Frame) 为574个氨基酸, 与双分病毒属( Partitivirus) 中部分病毒的RdRp (RNA dependent RNA polymerase) 氨基酸序列具有一定的同源性。推测此dsRNA可能为萝卜黄边病毒( radish yellow edge virus, RYEV) 感染所致。

关键词: 萝卜, 双链RNA, cDNA克隆, 核酸序列, 萝卜黄边病毒

Abstract: Some characters of unknown dsRNAs isolated from radish were identified. The dsRNAs could be detected from some radish plants and their offspring by dsRNAs analysis approach. The dsRNAswere separated into two fragments in polyacrylamide gel, named dsRNA1 and dsRNA2, respectively with their sizes of
approximately 118 kbp. Full-length cDNA of dsRNA1 was obtained by single primer amplification technique, and its size of 1 866 ntwas determined. RT-PCR also showed that dsRNA1 was of radish plant origin. The results of BLASTP suggested that the biggest putative ORF1 was distantly related to RNA dependent RNA polymerase encoded by some species of Partitivirus. The molecular aspects of dsRNAs, and some biological characters of plants, seem related to radish yellow edge virus (RYEV) previously described.

Key words: Radish, Double-stranded RNA, cDNA cloning, Sequence, Radish yellow edge virus

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