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作物学报 ›› 2014, Vol. 40 ›› Issue (06): 1027-1034.doi: 10.3724/SP.J.1006.2014.01027

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

一个快速响应干旱的F-box基因的克隆和表达分析

尹恒,余琴鸯,安利佳,李文利   

  1. 大连理工大学生命科学与技术学院, 辽宁大连 116024
  • 收稿日期:2013-09-26 修回日期:2014-01-12 出版日期:2014-06-12 网络出版日期:2014-03-24
  • 通讯作者: 李文利, E-mail: biolwl@dlut.edu.cn
  • 基金资助:

    本研究由辽宁省科技厅农业攻关项目(2011208001)资助。

Cloning and Expression Analysis of an F-box Gene (SiFBX) Rapidly Responsive to Drought Stress

IN Heng,YU Qin-Yang,AN Li-Jia,LI Wen-Li   

  1. School of Life Science & Biotechnology, Dalian University of Technology, Dalian 116023, China
  • Received:2013-09-26 Revised:2014-01-12 Published:2014-06-12 Published online:2014-03-24

摘要:

F-boxSkp1-Cullin1-F-box (SCF)型泛素连接酶E3的重要组成部分,在泛素化介导的蛋白质降解中选择性识别靶蛋白。本文从谷子苗期干旱胁迫条件下构建的转录组文库中克隆到与耐旱早期响应相关的F-box基因,命名SiFBX (GenBank登录号为KC252635.1)。该基因全长510 bp,编码170个氨基酸。蛋白质结构预测表明,该蛋白含有丰富的精氨酸、亮氨酸、丝氨酸,缺少跨膜结构域及信号肽序列。系统进化分析表明,该基因与已报道的EID1FBW4亲缘关系较近。在该基因上游1.9 kb序列处,预测到启动子的核心序列及与多种逆境胁迫相关的调控序列。荧光定量PCR分析表明,该基因分别在正常干旱、PEGABA诱导下,表达量出现显著变化。

关键词: 谷子, 干旱响应, F-box, 表达分析

Abstract:

F-box proteins, components of the Skp1-Cullin1-F-box (SCF) protein E3 ubiquitin ligase complex, serve as the variable component responsible for substrate recognition and recruitment in SCF-mediated proteolysis. The anti-drought relative gene of SiFBX (GenBank accession number KC252635.1) which belongs to the F-box super family was cloned from foxtail millet (Setaria italic). The full-length cDNA of SiFBX was 510 bp, which encoded 170 amino acid residues. Protein analysis and structure predication showed that it had a higher proportion of arginine (R), leucine (L), and serine (S) and a lack of trans-membrane domains and signal peptide. Phylogenetic analysis demonstrated that SiFBX had similarity with EID1 and FBW4. Many abiotic stress-related cis-acting elements and transcription factors were discovered in the 1.9 kb upstream region of SiFBX. The results of real-time PCR showed that there were remarkable changes in the expectation level of SiFBX for the treatments with PEG, water-withholding and ABA.

Key words: Setaria italica, Drought response, F-box protein, qRT-PCR

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