Cloning and sequencing of DREB2A gene and construction of its plant expression vector

A 1 050 bp DNA fragment which was amplified by RT-PCR using DREB2A gene special primer from Arabidopsis thaliana leaf treated by 200 mmol·L-1 NaCl was cloned into pUC18. The result of sequencing showed that the sequence of this fragment was 100% homologous with that of DREB2A gene in GenBank. After that DREB2A gene was cloned into plant expression vector cassette pCCE12 and pCC29. Thus two plant expression vectors, pCDRE12 and pCDR29A were constructed, in which DREB2A gene was controlled by E12 promoter and rd29A promoter respectively. Then the recombination plasmids were introduced into Agrobacterium LBA4404 by freezing -melting transformation method. This work provides a foundation for the transformation of DREB2A gene to plant mediated by Agrobacterium.