Transformation of Arabidopsis by Flag tagged PP2A

Eukaryotic protein kinases transfer a phosphate group from ATP to the hydroxyl group of Ser, Thr and Tyr residues, whereas protein phosphatases hydrolyze the phosphoester bond to give free phosphate and dephosphorylated protein. Protein phosphorylation and dephosphorylation are essential for the regulation of metabolism, cell division, development, growth, and stress response in all organisms. Protein phosphatase type 2A is a serine and threonine specific phosphatase. The enzyme is characterized by its multi-subunit structure that allow it to participate in a wide range of signaling cascades that regulate plant growth, defense and development. Protein phosphatase type 2A is made up of three subunits, a catalytic (C), scaffold (A) and regulatory (B) subunit. The regulatory subunit is divided into three groups called B, B' and B''. A particular subgroup of B' called B'φ (B'phi) appears to be crucial for the interaction between plants and microorganisms, especially mycorrhiza. B'φ has not been studied much and can also be important for regulating other processes. The model plant Arabidopsis thaliana (Arabidopsis) does not have the subunit B'φ, and the aim of this project is to i) clone B'φ from Solanum lycopersicum (tomato plant) ii) link the tomato gene with a FLAG tag and transform it into an Arabidopsis. Arabidopsis plants will be selected on BASTA for further cultivation and the presence of B'φ will be confirmed with PCR. Further experiments can provide information about the location of the protein in the cell, and interaction partners to compare the interacting proteins identified in Arabidopsis and tomato