Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2021802 | Protein Expression and Purification | 2007 | 8 Pages |
In this paper, we evaluated various parameters of culture condition affecting high-level soluble expression of human cyclin A2 in Escherichia coli BL21(DE3), and demonstrated that the highest protein yield was obtained using TB(no glycerol) + 0.5% glucose medium at 25 °C. By single immobilized metal ion affinity chromatography, we got highly purified human cyclin A2 with a yield ranged from 20 to 30 mg/L. By amyloid-diagnostic dye ThT binding and Fourier transform infrared spectroscopy, we observed a significant decrease in α-helix content and an increase in β-sheet structure in cyclin A2 inclusion body in comparison to its native protein, and confirmed the resemblance of the internal organization of cyclin A2 inclusion body and amyloid fibrils.