Cloning and analysis of a novel DREB gene of Glycine soja

The total RNA in leaf of Glycine soja was treated as cyclostyle. The degenerate primers were designed based on the conservative amino acids sequence of DREBP/AP2 domain from other dicotyledonous. A cDNA fragment of 191bp from G. soja was obtained by touchdown PCR and nestedPCR. Based on this sequence, the nest primers were designed, than 3and 5cDNA sequences were obtained by 3and 5 rapid amplification of cDNA ends PCR (RACEPCR), and the complete DREB gene was constructed in G. soja (designated as GsDREB). The whole sequence of GsDREB was 1191 bp open reading frame (ORF) encoding a 395 amino acids, without any intron. Amino acid analysis indicated that, the predicted GsDREB protein contained a Nterminal nuclear localization signal (NLS), and a putative AP2 DNAbinding domain containing 58 conserved amino acids. Alignment of multiple showed that GsDREB had much similarity with AtDREB2, so it might be a member of DREB2 subgroup of DREB family.