Cloning of Arabidopsis HIS1-3 gene and construction of its expression vector

The total RNA was extracted from Arabidopsis seedlings,the cDNA fragments of HIS1-3 gene were amplified by RT-PCR and lighted with pEASY-T1 Simple vector.The pEASY-T1-HIS1-3 was transformed into Trans1-T1 phage resistant chemically competent cells,the single masccline colony was screened and the plasmid was extracted.Then the restriction enzymes of Eco91 Ⅰ and Nco Ⅰ were used to completely double-digeste the pEASY-T1-HIS1-3 plasmid and the pCAMBIA2301 vector.The recombinant pCAMBIA2301-HIS1-3 vector was carried into the C58 strain of Agrobacterium tumefaciens by electrotransformation.Finally,the strains of Agrobacterium tumefaciens carrying HIS1-3 gene were obtained.The study provides the basis for studying the molecular mechanism of stress-tolerance of HIS1-3 gene and improving the stress-tolerance of crops.