Cloning of Arabidopsis thaliana AtMYB61 gene and the expression vector construction

Total RNA was extracted from Arabidopsis thaliana seedlings and used as the template to amplify the full length of cDNA of AtMYB61 gene by RT-PCR technology,and the gene fragment was subsequently cloned into plant pUCm-T vector.The results of bacterial colony PCR,enzyme analysis and cDNA sequencing confirmed that the Arabidopsis thaliana AtMYB61 gene was successfully cloned.AtMYB61 gene was cut completely from AtMYB61-pUCm-T vector by BstEⅡand BglⅡ,and the gene fragment was cloned into plant expression vector pCAMBIA2301.The results of bacterial colony PCR and enzyme analysis showed the successful construction of plant expression vector pCAMBIA2301-AtMYB61.In addition,the recombinant expression vector was carried into Agrobacterium tumefaciens by electrotransformation,and the strain of Agrobacterium tumefaciens carrying AtMYB61 gene was obtained.The study provides a basis for improving the resistance of transgenic plants and further exploring the molecular mechanism of AtMYB61 gene.