Plasmid DNA purification via the use of a dual affinity protein
Forde, G.M. (2008) Plasmid DNA purification via the use of a dual affinity protein. In Zachariou, M. (Ed.) Affinity Chromatography : Methods and Protocols [2nd ed.]. Humana Press (Springer), pp. 275-284.
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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|Item Type:||Book Chapter|
|Divisions:||Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > QUT Faculties and Divisions > Science & Engineering Faculty
|Deposited On:||09 Feb 2015 02:46|
|Last Modified:||12 Feb 2015 05:46|
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