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林业科学 ›› 2010, Vol. 46 ›› Issue (4): 37-42.doi: 10.11707/j.1001-7488.20100406

• 论文 • 上一篇    下一篇

毛竹Stowaway-like MITEs转座子的分离与分析

钟浩1 周明兵1 白有煌2 汤定钦1   

  1. 1.浙江林学院 浙江省现代森林培育技术重点实验室 临安 311300; 2.浙江大学生命科学学院生物信息系 杭州 310058
  • 收稿日期:2009-08-31 修回日期:1900-01-01 出版日期:2010-04-25 发布日期:2010-04-25
  • 通讯作者: 汤定钦

Isolation and Analysis of Stowaway-like MITEs from DNA Libraries Enriched in Phyllostachys edulis

Zhong Hao1,Zhou Mingbing1,Bai Youhuang2,Tang Dingqin1   

  1. 1. Zhejiang Key Laboratory for Modern Silvicultural Technology Zhejiang Forestry University Lin’an 311300;2. Department of Bioinformatics, College of Life Sciences, Zhejiang University Hangzhou 310058
  • Received:2009-08-31 Revised:1900-01-01 Online:2010-04-25 Published:2010-04-25

摘要:

微型颠倒重复序列(miniature inverted repeat transposable elements, MITEs)是一类对基因组进化和基因表达有重要调节作用的转座子。为分析MITEs在毛竹基因组中的分布特性,借鉴Fast Isolation by AFLP of Sequences Containing repeats ( FIASCO)方法,首次构建Stowaway-like MITEs富集文库。随机挑取21个克隆,测序发现2个含有Stowaway-like MITEs序列 (Stow-Ph1Stow-Ph2),阳性率为9.52%。Stow-Ph1Stow-Ph2的长度分别为260和258 bp,具有Stowaway-like MITEs典型的末端颠倒重复序列(terminal inverted repeats, TIRs)和靶位点重复序列(target site duplication, TSD)。Stow-Ph1Stow-Ph2与水稻的Stow-Os8(FJ266024)的同源性分别为46.4%和52.6%。Stow-Ph1Stow-Ph2与水稻16类Stowaway-like MITEs的TIRs前20 bp的相似性均高于50%,表明毛竹的Stowaway-like MITEs在TIRs上是高度保守的。

关键词: 毛竹, 磁珠富集, 转座子, TIRs, Stowaway-like MITEs

Abstract:

Transposable elements were a kind of DNA fragments movable in genomes. Miniature inverted repeat transposable elements (MITEs) were a class of short, widespread transposable elements with high copy number which played an important role in evolution of genome and regulation of gene expression. In order to analyze the distribution and characteristics of MITEs in Phyllostachys edulis genome, DNA library enriched for Stowaway-like MITEs was firstly constructed by the method of FIASCO (Fast Isolation by AFLP of Sequences Containing repeats). Totally 21 recombinant clones were randomly isolated with PCR method and then sequenced. Two clones (Stow-Ph1 and Stow-Ph2) were identified as Stowaway-like MITEs. The rate of positive clones was 9.52%. The fragment lengths of Stow-Ph1 and Stow-Ph2 were 260 bp and 258 bp, respectively. They had typical TIRs (terminal inverted repeats) and TSD (target site duplication) of Stowaway-like MITEs. The identity was 46.4% between Stow-Ph1and Stow-Os8(FJ266024), and 52.6% between Stow-Ph2and Stow-Os8. The identities among Stow-Ph1,Stow-Ph2and 16 rice Stowaway-like MITEs were above 50% based on the first 20 bp of their TIRs, which reflected that the TIRs of Stow-Ph1 and Stow-Ph2 were highly conserved.

Key words: Phyllostachys edulis, enrichment by magnetic beads, transposon, TIRs, Stowaway-like MITEs