作物学报 ›› 2013, Vol. 39 ›› Issue (08): 1352-1359.doi: 10.3724/SP.J.1006.2013.01352

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇



  1. 1河北农业大学农学院,河北保定071000;2河北农业大学资源与环境科学学院,河北保定071000
  • 收稿日期:2013-02-04 修回日期:2013-04-22 出版日期:2013-08-12 网络出版日期:2013-05-20
  • 通讯作者: 常金华, E-mail: changjinhua@hebau.edu.cn
  • 作者简介:常金华, E-mail: changjinhua@hebau.edu.cn
  • 基金资助:


Cloning and Expression Analysis of SbDREB2 Gene from Sorghum bicolor

XIE Deng-Lei1,CUI Jiang-Hui2,CHANG Jin-Hua1,*   

  1. 1 College of Agronomy, Agricultural University of Hebei, Baoding 071000, China; 2 College of Resources and Environment Science, Agricultural University of Hebei, Baoding 071000, China
  • Received:2013-02-04 Revised:2013-04-22 Published:2013-08-12 Published online:2013-05-20
  • Contact: 常金华, E-mail: changjinhua@hebau.edu.cn
  • About author:常金华, E-mail: changjinhua@hebau.edu.cn


干旱应答元件结合蛋白(DREB)在植物非生物逆境胁迫中调节下游一系列抗逆基因的表达。本研究利用电子克隆和RT-PCR方法从高粱中克隆到1DREB类基因SbDREB2,该基因ORF 789 bp,推测编码蛋白含262个氨基酸残基,相对分子质量28.6 kD,理论等电点为5.52,在DNA序列内包含1740 bp的内含子,符合GT-AG剪接规则。氨基酸序列分析表明,该蛋白在82~145区含有DREB类转录因子家族特有的AP2保守结构域,与玉米DREB2A及水稻DREB1蛋白相似度分别为84%69%。成功构建了原核表达载体pET28a-SbDREB2,经IPTG诱导获得32.5 kD左右蛋白,与理论值一致。Real-time PCR表达特性分析显示,该基因为组成型表达,在根、茎、叶中均表达,根中表达量约是茎中的2.5倍;受干旱、高盐和外源ABA的强烈诱导,但对低温几乎没有响应。

关键词: 高粱, SbDREB2, 原核表达, 胁迫处理, 荧光定量PCR


DREBs play important roles in regulating the expression of downstream genes in response to a variety of abiotic stresses. In this paper, a DREB-like gene, named SbDREB2, was assembled by searching sorghum EST and genome databases. The SbDREB2 gene was cloned from salinity-stressed sorghum seedling by RT-PCR. SbDREB2 contains a 789 bp complete open reading frame (ORF) which encodes a peptide of 262 amino acids. The predicted molecular weight and isoelctric point of SbDREB2 are 28.64 kD and 5.52, respectively. There is a 740 bp intron in the DNA sequence of SbDREB2. The amino acids analysis indicated that the predicted protein sequence contained a typical AP2 DNA-binding domain in the 82–145 regions. Multiple sequences alignment revealed that SbDREB2 shared 84% and 69% sequence similarities with Zea mays DREB2A and Oryza sativa DREB1, respectively. Prokaryotic expression vector pET28a-SbDREB2 was established and transformed BL21 (DE3) into E. coli after IPTG induction, showing a successful gene expression. The expression pattern analysis carried out by quantitative real-time PCR indicated that SbDREB2 was constitutively expressed in various tissues of sorghum, and was strongly up-regulated under high salinity, drought and exogenous application of abscisic acid (ABA). However, the expression of SbDREB2 was not affected by low temperature.

Key words: Sorghum, SbDREB2, Prokaryotic expression, Abiotic stress, Quantitative real-time PCR

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