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林业科学 ›› 2009, Vol. 12 ›› Issue (10): 69-73.doi: 10.11707/j.1001-7488.20091012

• 论文 • 上一篇    下一篇

休眠期珙桐种子cDNA文库构建及EST分析*

齐刚1,2 苏智先2 李劲涛3 阮期平3   

  1. 1.西华师范大学生命科学学院 南充637002; 2.四川省生态学与环境保护技术重点实验室 绵阳621000;3.绵阳师范学院分子生物学与生物制药重点实验室 绵阳621000
  • 收稿日期:2008-08-06 修回日期:1900-01-01 出版日期:2009-10-25 发布日期:2009-10-25
  • 通讯作者: 苏智先
  • 基金资助:
     

Construction of cDNA Library and Analysis of the Expressed Sequenced Tags (ESTs) Characteristics of Dormant Seeds of Davidia involucrata

Qi Gang1,2,Su Zhixian2,Li Jingtao3,Ruan Qiping3   

  1. 1.School of Life Sciences, China West Normal University Nanchong 637002; 2.Key Laboratory of Ecology and Environmental Protection Technology of Sichuan Province Mianyang 621000;3.Key Laboratory for Molecular Biology and Biopharmaceuticals of Mianyang Normal University Mianyang 621000
  • Received:2008-08-06 Revised:1900-01-01 Online:2009-10-25 Published:2009-10-25
  • Supported by:
     

摘要:

以休眠期的珙桐种子为材料,利用SMARTTM cDNA Library Construction Kit构建珙桐种子的cDNA文库。结果表明: 该文库的滴度为1.5×106 pfu·mL-1,重组率为98%, cDNA文库插入片段大小主要集中在0.5~1.0 kb。随机测序获得148个EST序列,拼接成127个uniEST,经网上BlastN及BlastX分析,其中75个序列是已知功能的基因标签,相似性较高,而32个序列在GenBank中相似性较低,13个序列在GenBank中没有匹配。这些数据有助于对珙桐种子的相关功能蛋白及分子机制的研究。

关键词: 珙桐, 种子, 休眠, cDNA文库, 表达序列标签

Abstract:

A cDNA library was constructed with dormant Davidia involucrata seeds by using SMART cDNA Library Construction Kit. The library had a titer of 1.5×106 pfu·mL-1, with recombinant rate of 98%. The inserted cDNA fragments ranged mainly from 0.5 to 1.0 kb. Total 148 ESTs were obtained from randomly sequencing, and 127 uniESTs were obtained by contiganalsis. By searching on line with BlastN and BlastX, 75 uniESTs were determined to be the tags of genes with putative function and high similarity, 32 uniESTs had low similarity and 13 uniEST had no significant similarity to any protein or DNA sequence in the databases. The present ESTs provide a valuable resource for further research on similarity protein function and molecule mechanism of Davidia involucrata seeds.

Key words: Davidia involucrata, seed, dormancy, cDNA library, ESTs

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