园艺学报 ›› 2008, Vol. 35 ›› Issue (4): 493-500.

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赵 英;牛建新*   

  1. (石河子大学农学院园艺系,新疆石河子,832003)
  • 收稿日期:2007-08-20 修回日期:2008-02-27 出版日期:2008-04-25 发布日期:2008-04-25
  • 通讯作者: 牛建新

Cloning of the Terminal Regions of Apple stem pitting virus in Pear Trees Using 3' and 5' RACE

ZHAO Ying and NIU Jian-xin*   

  1. (Department of Horticulture, Agricultural College of Shihezi University, Shihezi, Xinjiang 832003, China)
  • Received:2007-08-20 Revised:2008-02-27 Online:2008-04-25 Published:2008-04-25
  • Contact: NIU Jian-xin

摘要: 采用RACE技术对库尔勒香梨苹果茎痘病毒进行研究,获得了梨树苹果茎痘病毒5′和3′末端序列。研究结果表明,本试验采用的3′和5′末端快速扩增技术(3′RACE和5′RACE技术)能很好地扩增苹果茎痘病毒的末端序列,分别获得了长度为329 bp和367 bp的片段,通过与 NCBI 核酸数据库中已经发表的序列NC_003462进行比对分析,发现梨树ASPV 3′末端含有131个核苷酸非编码序列,具有 poly(A)尾,5′末端含有34个核苷酸非编码序列,其同源性与已发表的序列分别为78%和84%,为获得梨树ASPV病毒全长基因组和基因组功能结构分析奠定了基础。

关键词: 梨, 苹果茎痘病毒, 3′和5′末端快速扩增法, poly(A)尾

Abstract: The 5' and 3' terminal regions of Apple stem pitting virus (ASPV) in tree of Pyrus sinkiangensis ‘Kuerle’pear were cloned using rapid amplification of cDNA ends (RACE) technique. The results showed that the techniques of 3' and 5' RACE could amplify the terminal regions of ASPV in pear trees effectively, and the 329 bp and 367 bp segments of ASPV were obtained. The 3' terminal region of ASPV in Kuerle pear contained 131 nt of non-coding sequence and a poly (A) tail, and the 5' terminal region contained 34 nt of non-coding sequence. They shared 78% and 84% nucleotide acid identity with the sequence of NC_003462 in NCBI nucleotide acid database, respectively. This study laid a sound foundation for obtaining the full sequence and analyzing the function and structure of genome of ASPV in pear trees.

Key words: pear, Apple stem pitting virus, rapid amplification of 3' and 5' terminal regions, poly (A) tail