Cabanne C, Noubhani A M, Hocquellet A, Dole F, Dieryck W, Santarelli X
Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Apr 15;818(1):23-7. doi: 10.1016/j.jchromb.2004.10.023.
The enhanced green fluorescent protein (EGFP) was over-expressed in Escherichia coli as inclusion bodies to increase its quantity and to facilitate its purification. Insoluble EGFP has been purified on Q Hyper Z matrix by expanded bed adsorption after solubilization in 8 M urea. The adsorption was made in expanded bed mode to avoid centrifugation. EBA-column refolding was done by elimination of urea and elution with NaCl. The EGFP was obtained as a highly purified soluble form with similar behavior in fluorescence and electrophoresis as native EGFP.
增强型绿色荧光蛋白（EGFP）在大肠杆菌中作为包涵体过度表达，以增加其产量并便于纯化。不溶性EGFP在8 M尿素中溶解后，通过扩张床吸附在Q Hyper Z基质上进行纯化。吸附在扩张床模式下进行以避免离心。通过去除尿素并用NaCl洗脱进行EBA柱复性。获得的EGFP为高度纯化的可溶性形式，在荧光和电泳方面与天然EGFP具有相似的行为。
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