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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (08): 1352-1359.doi: 10.3724/SP.J.1006.2013.01352

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2013-08-12 Published:2013-05-20
  • Contact: 常金华, E-mail: changjinhua@hebau.edu.cn
  • About author:常金华, E-mail: changjinhua@hebau.edu.cn

Abstract:

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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