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.
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© 2003 Humana Press Inc., Totowa, NJ
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Sun, X.Z., Nguyen, J., Momand, J. (2003). Purification of Recombinant p53 from Sf9 Insect Cells. In: Deb, S., Deb, S.P. (eds) p53 Protocols. Methods in Molecular Biology, vol 234. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-408-5:17
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DOI: https://doi.org/10.1385/1-59259-408-5:17
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-1-58829-106-6
Online ISBN: 978-1-59259-408-5
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