Soluble Expression of Plasmodium falciparum Glutamate Dehydrogenase in Escherichia coli, and its Purification and Identification

Abstract

Abstract:

Objective To make soluble expression of Plasmodium falciparum (FCC1/HN) glutamate dehydrogenase(GDH) in Escherichia coli, purification and immunocompetence identification of the recombinant non-fusion GDH. Methods The GDH gene was cloned into prokaryotic expression vector pET23(a) to form recombinant expression vector pET23(a)/GDH. pET23(a)/GDH was transformed into E.coli BL21(DE3). Induced by IPTG(isopropyl-beta D-thiogalactoside), GDH was highly expressed in the supernatant after sonication. The soluble recombinant GDH was purified by Source-Q and Source-S chromatography. Enzyme-linked immunosorbent assay and Western blotting were carried out to identify the immunocompetence of the purified product. Results SDS-PAGE analysis showed that the soluble GDH protein accounted for approximately 15% of the total bacterial protein. By two-step ion-exchange chromatography, the purity of GDH reached more than 90% and the GDH possessed high antigenicity. Conclusion The soluble expression of GDH results in an integral three-dimensional structure epitope with high biological activity.

Key words: Plasmodium falciparum, glutamate dehydrogenase, gene expression, Western blotting

LIYan;NINGYun-shan;DONGWen-qi;LIMing. Soluble Expression of Plasmodium falciparum Glutamate Dehydrogenase in Escherichia coli, and its Purification and Identification[J]. , 2004, 22(2): 8-97.

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Soluble Expression of Plasmodium falciparum Glutamate Dehydrogenase in Escherichia coli, and its Purification and Identification

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