Four methods were compared to purify Fo from porcine h ear t mitochondria. The best results were obtained by the following method: after re moving F1-ATPase with NaBr incubation from submitochondrial FoF1-ATPase, Fo was solubilized with CHAPS and purified by sucrose density gradient centri fugation. SDS-PAGE with silver staining showed about 85% purity of the isolated Fo and 9 different subunits including b, OSCP, d, a, e, F6, IF1, A6L and c. The purified Fo was then incorporated into asolectin liposomes, the reconst ituted Fo showed higher H+ translocation activity and after Fo was reconst ituted with F1-ATPase, the resulted FoF1-ATPase complex exhibited high A TP hydrolysis activity and high sensitivity to oligomycin. The results provide e vidence for successful purification, reconstitution of Fo with high H+ trans location activity and its relationship with phospholipids.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.