利用扩张床阴离子交换色谱法对在大肠杆菌中作为包涵体过量表达的增强型绿色荧光蛋白进行纯化及柱上复性。
Purification and on-column refolding of EGFP overexpressed as inclusion bodies in Escherichia coli with expanded bed anion exchange chromatography.
作者信息
Cabanne C, Noubhani A M, Hocquellet A, Dole F, Dieryck W, Santarelli X
机构信息
Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
出版信息
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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