title: Purification of recombinant p53 from Sf9 insect cells.
abstract: We describe a method for purifying recombinant p53 from baculovirus infected cells in one step by anion exchange chromatography. The p53 is full-length with no flanking sequences and its expression is driven by the baculovirus polyhedron promoter. We also describe how to concentrate the p53 up to 0.9 mg/mL. By gel filtration analysis, we demonstrate that 20% of the p53 forms a tetramer, and 80% forms a monomer. In a DNA binding assay known as the electromobility shift assay, the purified p53/DNA complex forms a single band the gel. This simple procedure should be useful for investigations into the biochemistry of the p53 protein.
authors.affiliation: Department of Chemistry and Biochemistry, California State University at Los Angeles, USA.
authors.forename: Xiu Zhu
authors.initials: XZ
authors.lastname: Sun
authors.forename: John
authors.initials: J
authors.lastname: Nguyen
authors.forename: Jamil
authors.initials: J
authors.lastname: Momand
mesh_terms: D000818:Animals; D002460:Cell Line; D002850:Chromatography, Gel; D004247:DNA / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007313:Insecta; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D004247:DNA