Plasmid DNA purification via the use of a dual affinity protein


Autoria(s): Forde, G.M.
Contribuinte(s)

Zachariou, M.

Data(s)

2008

Resumo

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.

Identificador

http://eprints.qut.edu.au/81619/

Publicador

Humana Press (Springer)

Relação

http://www.springer.com/life+sciences/cell+biology/book/978-1-58829-659-7

DOI:10.1007/978-1-59745-582-4_18

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.

Fonte

School of Chemistry, Physics & Mechanical Engineering; Science & Engineering Faculty

Tipo

Book Chapter