Cloning of the Protein Phosphatase AtPP2C Gene of Arabidopsis thaliana and Construction of Plant Express Vector

According to the cDNA sequence of AtPP2C gene(AT1g07430) in GenBank a pair of primer was designed and synthesized.Using the total RNA of NaCl solution treated Arabdopsis thaliana seedling as template,the full length of cDNA of AtPP2C gene was amplified through method of RT-PCR.It was cloned into pDONR201 vector using BP reaction of Gateway cloning technology,and the entry clone was gained.DNA sequencing indicated that the cloned fragment showed 100% identity to the sequence of the gene in GenBank.Using the LR reaction of Gateway cloning technology,the plant express vector pLEELA was ligated with the entry clone and got the AtPP2c gene express vector pLAT35S regulated by 35S promoter.