Prokaryotic expression of Trx-apoptosis-inducing factor-defected mitochondria localization sign fusion protein and its effect on cell-free system of leukemia K5 62 cells / 肿瘤

ABSTRACT

Objective: To express the fusion protein-thioredoxin-apoptosisinducing factor-defected mitochondria localization sign (Trx-DMLS-AIF) in prokaryotic cells and to detect its effect on cell-free system of leukemia K562 cells. Methods: The recombinant plasmid expressing Trx-DMLS-AIF fusion protein was established and transformed into E.coli BL21. The expression of Trx-DMLS-AIF fusion protein was induced by isopropyl β-D-1-thiogalactopyranoside (IPTG) and purified by Ni-NTA Spin Columns. The cell-free system of leukemia K562 cells was established. The effect of Trx-DMLS-AIF fusion protein on cell-free system of K562 cells was detected by acridine orange (AO) staining. The effect of plasmosin on the ability of Trx-DMLS-AIF fusion protein entering into nuclei of K562 cells was detected by immunofluorescent staining. Results: The expression of Trx-DMLS-AIF fusion protein was successfully induced by IPTG, and the purified Trx-DMLS-AIF fusion protein was obtained. The result of AO staining showed that Trx-DMLS-AIF fusion protein could induce the nucleus apoptosis in cell-free system of K562 cells. The result of immunofluorescent staining showed that the plasmosin of K562 cells could prevent Trx-DMLS-AIF fusion protein from entering into the nuclei of K562 cells. Conclusion: The expression of Trx-DMLS-AIF fusion protein is successfully induced. TrxDMLS-AIF can induce the nucleus apoptosis of K562 cells, and the plasmosin of K562 cells can prevent Trx-DMLS-AIF fusion protein from entering into the nuclei of K562 cells.