生物多样性 ›› 2012, Vol. 20 ›› Issue (1): 51-58.DOI: 10.3724/SP.J.1003.2012.08168

• 研究报告 • 上一篇    下一篇

商城肥鲵二碱基重复和四碱基重复微卫星DNA的结构特征及对筛选效率的影响

王慧1, 张保卫1,*(), 史文博1, 骆侠1, 周立志1, 韩德民1, 常青2   

  1. 1 安徽大学生命科学学院, 合肥 230039
    2 南京师范大学生命科学学院, 南京 210046
  • 收稿日期:2011-09-02 接受日期:2011-10-16 出版日期:2012-01-20 发布日期:2012-02-14
  • 通讯作者: 张保卫
  • 作者简介:*E-mail: zhangbw@ahu.edu.cn
  • 基金资助:
    国家自然科学基金(30800116);生物多样性与生物技术江苏省重点实验室开放基金(B200712-01);安徽大学青年骨干教师计划

Structural characteristics of di-nucleotide/tetra-nucleotide repeat microsatellite DNA in Pachyhynobius shangchengensisgenomes and its effect on isolation

Hui Wang1, Baowei Zhang1,*(), Wenbo Shi1, Xia Luo1, Lizhi Zhou1, Demin Han1, Qing Chang2   

  1. 1 School of Life Sciences, Anhui University, Hefei 230039
    2 School of Life Sciences, Nanjing Normal University, Nanjing 210046
  • Received:2011-09-02 Accepted:2011-10-16 Online:2012-01-20 Published:2012-02-14
  • Contact: Baowei Zhang

摘要:

两栖类有尾目物种的微卫星分离中的筛选成功率常常较低。为探索微卫星结构对筛选效率的影响, 本研究通过AFLP快速分离法(fast isolation by AFLP of sequences containing repeats, FIASCO)对商城肥鲵(Pachyhynobius shangchengensis)二碱基重复类型和四碱基重复类型微卫星进行分离, 并对微卫星序列进行了分析。研究中发现二碱基微卫星位点多以微卫星DNA家族形式存在, 并因此导致了微卫星位点分离较低的筛选率; 在四碱基重复的微卫星位点中未发现微卫星DNA家族的存在。对研究中得到的3个微卫星DNA家族的分析发现, 同一家族的上、下游侧翼序列变异程度存在差异; 毗邻微卫星重复单元区的侧翼序列碱基变异程度较高, 而较远处的区段则相对保守。这些结构特征可能反映出微卫星DNA家族在演化中的复杂性。本文的研究结果提示在两栖动物的一些类群中, 微卫星的筛选必须考虑微卫星DNA家族的影响, 选取适宜的碱基重复类型将是决定筛选效率的关键。

关键词: 微卫星结构, 微卫星DNA家族, Pachyhynobius shangchengensis, 筛选效率

Abstract:

A low success ratio is often encountered in the microsatellite isolation from the species of Caudata. In the present study, di-nucleotide/tetra-nucleotide repeat microsatellite loci were isolated following the FLASCO (fast isolation by AFLP of sequences containing repeats) approach and the sequences were analyzed for the purpose of learning how microsatellite structure impacted on isolation efficiency. We found that most di-nucleotide repeat microsatellite loci were involved in the microsatellite DNA family, which resulted in low isolation efficiency. In contrast, no microsatellite DNA family members were found in the tetra-nucleotide repeat microsatellite loci. On the basis of analysis of the three microsatellite DNA families found in the present study, there were significant differences in the extent of variation between the upstream and downstream flanking sequences in same microsatellite DNA family. This variation was the highest in the sequence sections adjacent to the core repeated units, but more conservative in other regions, which might reflect the complexity of the microsatellite DNA family evolution. These results suggest that in the process of microsatellite development for Caudata, we must pay attention to interference from microsatellite DNA family evolution. The choice of appropriate nucleotide repeats is key to obtaining a high isolation efficiency.

Key words: microsatellite, structure characteristics, microsatellite DNA family, Pachyhynobius shangchengensis, isolation efficiency