Molecular Cloning and Characterization of a Novel Gene Encoding DREB Protein from Buchloe dactyloides (Nutt.) Engelm

Yan Sun, Yan Li, Xiaoyan Hu, Qingchuan Yang, Junmei Kang and Tiejun Zhang

Journal of Agricultural Science, 2012, vol. 4, issue 1, 12

Abstract: A 209bp expressed sequence tag (EST) was obtained with the degenerate primer technology, then based on the sequence of EST, the full-length cDNA of 1050 nucleotides was cloned from Buffalo grass by rapid amplification of cDNA ends (RACE). It was designated as BdDREB2, encoding a protein of 255 amino acids. The protein molecular weight was 28.39kDa, and the theoretical isoelectric point was 5.70. The BdDREB2 probably localized in nucleus and did not have signal peptide. A typical AP2/EREBP conserved domain was found in the coding region. The amino acid sequence compared by blast revealed high homology with DREB proteins of other plants, especially with the Buchloe-dactyloi DREB protein. The semi-quantitative analysis of expression of transcription showed- the expression of BdDREB2 gene reached the maximum after being treated by 20%PEG for 6h; BdDREB2 gene has the highest expression level in the root; the expression was significantly increased with the stress of drought (20% PEG) and high salt (3% NaCl), but not obvious with the stress of low temperature. Plant expression vector was constructed and transformed into the tobacco by the Agrobacterium-mediated methods. PCR amplification with the transgenic tobacco genome DNA indicated that the BdDREB2 gene had integrated in tobacco genome. RT-PCR showed that the BdDREB2 gene can be transcribed into mRNA in tobacco.

Date: 2012
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