Summary
Methods are presented for the production, affinity purification and analysis of plasmid DNA (pDNA). Batch fermentation is used for the production of the pDNA, and expanded bed chromatography, via the use of a dual affinity glutathione S-transferase (GST) fusion protein, is used for the capture and purification of the pDNA. The protein is composed of GST, which displays affinity for glutathione immobilized to a solid-phase adsorbent, fused to a zinc finger transcription factor, which displays affinity for a target 9-base pair sequence contained within the target pDNA. A Picogreen™ fluorescence assay and/or anx ethidium bromide agarose gel electrophoresis assay can be used to analyze the eluted pDNA.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Forde, G.M. (2008). Plasmid DNA Purification Via the Use of a Dual Affinity Protein. In: Zachariou, M. (eds) Affinity Chromatography. Methods in Molecular Biology™, vol 421. Humana Press. https://doi.org/10.1007/978-1-59745-582-4_18
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DOI: https://doi.org/10.1007/978-1-59745-582-4_18
Publisher Name: Humana Press
Print ISBN: 978-1-58829-659-7
Online ISBN: 978-1-59745-582-4
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