Manufacture of dsDNA Microarrays of DRE Cis-acting Element

DRE cis-acting element which could specifically bind DREB transcription factors plays important roles in inducing gene expression involved in dehydration,low-temperature,and high-salinity stresses.Double-stranded DNA microarrys could very effectively detect the interaction between DNA-cis-acting elements and protein-transcription factors in regulation of gene expression,and also could availably analyze the binding interaction of bio-molecular.In this study,we designed and chemically synthesized a hairpin structure single-stranded DNA probe according to the nuclear sequence of DRE cis-acting element.Furthermore,we adopted Taq DNA polymerase in the elongation system and optimized the new on-chip reaction conditions.Our experiments showed that the elongation temperature of 50℃,the Mg²⁺ concentration of 2.5 mmol/L and single-stranded,non-denatured probe are the optimal conditions in elongation with Taq DNA polymerase.These results provide further steps toward a better understanding of interaction mechanisms between DRE cis-acting element and DREB transcription factors.