Recombinant Schizosaccharomyces pombe Major serine/threonine-protein phosphatase PP2A-2 catalytic subunit (ppa2)

To create the recombinant Schizosaccharomyces pombe ppa2 protein, the gene fragment encoding the full-length Schizosaccharomyces pombe ppa2 protein (1-322aa) is cloned into an expression vector. The expression vector is carefully engineered with an N-terminal 10xHis-tag and a C-terminal Myc-tag to facilitate the purification and detection of the recombinant protein. Following the construction of the recombinant expression vector, it is introduced into competent E. coli cells. The transformed cells are then selected on agar plates containing specific antibiotics corresponding to the expression vector's selection marker. Subsequently, the selected E. coli cells are cultured under optimal conditions to ensure efficient protein expression. Induction of protein expression is achieved using a suitable inducer, such as IPTG. Once induced, the E. coli cells are harvested by centrifugation and cell lysis is performed to release the cellular contents, including the recombinant ppa2 protein. The purity of the recombinant ppa2 protein is evaluated through SDS-PAGE analysis, confirming a purity level exceeding 85%.