Key Laboratory of Agrobiotechnology, College of Life Sciences, Shihezi University, Shihezi 832003, PR China.
School of Life Sciences, University of Science and Technology of China, Hefei 230027, PR China.
Z Naturforsch C J Biosci. 2013 Nov-Dec;68(11-12):499-508.
Olimarabidopsis pumila is a close relative of the model plant Arabidopsis thaliana but, unlike A. thaliana, it is a salt-tolerant ephemeral plant that is widely distributed in semi-arid and semi-salinized regions of the Xinjiang region of China, thus providing an ideal candidate plant system for salt tolerance gene mining. A good-quality cDNA library was constructed using cap antibody to enrich full-length cDNA with the gateway technology allowing library construction without traditional methods of cloning by use of restriction enzymes. A preliminary analysis of expressed sequence tags (ESTs) was carried out. The titers of the primary and the normalized cDNA library were 1.6 x 10(6) cfu/mL and 6.7 x 10(6) cfu/mL, respectively. A total of 1093 clones were randomly selected from the normalized library for EST sequencing. By sequence analysis, 894 high-quality ESTs were generated and assembled into 736 unique sequences consisting of 72 contigs and 664 singletons. The resulting unigenes were categorized according to the gene ontology (GO) hierarchy. The potential roles of gene products associated with stress-related ESTs are discussed. The 736 unigenes were similar to A. thaliana, A. lyrata, or Thellungiella salsuginea. This research provides an overview of the mRNA expression profile and first-hand information of gene sequence expressed in young leaves of O. pumila.
野生拟南芥是模式植物拟南芥的近亲,但与拟南芥不同的是,它是一种耐盐的短命植物,广泛分布于中国新疆的半干旱和半盐化地区,因此为耐盐基因挖掘提供了理想的候选植物系统。使用帽抗体构建了高质量的 cDNA 文库,通过 gateway 技术富集全长 cDNA,无需使用传统的酶切克隆方法。对表达序列标签 (EST) 进行了初步分析。初级和归一化 cDNA 文库的滴度分别为 1.6 x 10(6) cfu/mL 和 6.7 x 10(6) cfu/mL。从归一化文库中随机挑选了 1093 个克隆进行 EST 测序。通过序列分析,共生成 894 条高质量 EST,并组装成 736 个独特序列,包括 72 个连续序列和 664 个单序列。根据基因本体论 (GO) 层次结构对生成的基因进行分类。讨论了与应激相关 EST 相关的基因产物的潜在作用。736 个基因与拟南芥、拟南芥或盐芥相似。本研究概述了野生拟南芥幼叶中 mRNA 表达谱和基因序列的第一手信息。