Cloning of Transcription Factor DREB1A Gene and Construction of Plant Expression Vector by Gateway Clone Technique

Objective:To research the functional the DREB1A gene in plant tolerance to osmotic stress and to develop the constructing way of the plant expression vector by Gateway technology.Methods:A pair of primer was designed according to the mRNA sequence of DREB1A gene in GenBank databases,DREB1A of Arabidopsis thaliana was cloned.Two pair of primers containing attB adapter were designed separately by Gateway cloning technology.Platinum pfx DNA polymerase which can reduce the non-specific binding greatly was used during two PCR in which adding B sequence to the cloned gene.By the BP recombination reaction,the PCR product containing attB was transfered to an attP-containing donor vector to create an entry clone.Finally,DREB1A gene was shutted into pH2GW7 vector by LR recombination reaction.Results:The plant expression vector of pH2GW7-DREB1A was successfully constructed by identification.Conclusion:The results showed that it is easy to construct a plant expression vector by Gateway cloning technology,and it provides basic information for genetic transformation with this gene.