Construction of Plant Expression Vector of Heat Shock Factor Gene AtHsfAla from Arabidopsis

[Objective] Heat shock factors (HSFs) are the major transcription factors of eukaryotic heat shock responses. This study aims to investigate the adversity stress tolerance functions of Arabidopsis heat shock factor AtHsfA1a,which has important significance for in-depth understanding of adversity stress tolerance mechanisms of plants and further utilization of heat shock factor genes. [Method] Genomic DNA of Arabidopsis was extracted with CTAB method and purified to obtain Arabidopsis DNA samples for in vitro site-specific recombination cloning(Gateway cloning) to construct plant expression vector of heat shock factor AtHsfA1a. Firstly,donor vector pDONR 201/AtHsfA1a was constructed based on attB and attP site-specific recombination method (BP reaction),to identify E. coli transformants harboring correct sequence of AtHsfA1a by sequencing; secondly,plant expression vector pB7WG2/AtHsfA1a overexpressing Arabidopsis heat shock factor AtHsfA1a was constructed based on attL and attR site-specific recombination method (LR reaction),to screen E. coli transformants harboring target plasmid. [Result] Plant expression vector of Arabidopsis heat shock factor gene AtHsfA1a was constructed successfully. [Conclusion] This study not only provided experimental materials for acquiring transgenic plants overexpressing heat shock transcription factor AtHsfA1a,but also laid the foundation for further investigation of the diversity of adversity stress tolerance functions regulated by HSFs.