1. Shaoyang Medical College, Shaoyang, Hunan 422000, China; 2. Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Insititution of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, China 在期刊界中查找 在百度中查找 在本站中查找
2. Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Insititution of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, China 在期刊界中查找 在百度中查找 在本站中查找
2. Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Insititution of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, China 在期刊界中查找 在百度中查找 在本站中查找
2. Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Insititution of Pathogenic Biology, University of South China, Hengyang, Hunan 421001, China 在期刊界中查找 在百度中查找 在本站中查找
[Objective] This study aimed to clone the ferritin gene Abferritin from Acinetobacter baumannii and identify its anti-oxidative activity. [Methods] Relative expression of Abferritin under oxidative stress was analyzed by real-time PCR. The gene encoding sequence of Abferritin was inserted into the pET28a vector to generate the pET28a-Abferritin recombinant plasmid. This plasmid was transformed into the E. coli BL21(DE3) to create transformed strain of BL/pET28a-Abferritin. The Abferritin protein was expressed by IPTG induction and was purified by Ni2+-affinity chromatography. Kinetics of Fe2+ oxidation which catalyzed by Abferritin protein was determined by spectrophotometric analysis. Anti-oxidative activity of Abferritin was examined by the radical scavenging assay. Survival rations of the recombinant and control E. coli under the oxidative stress of H2O2 were also measured. [Results] Expression level of Abferritin was up-regulated in A. baumannii under oxidative stress. Abferritin protein was expressed in E. coli BL21(DE3) and was purified successfully. Ferroxidase activity assay demonstrated that the Abferritin protein could convert Fe2+ to Fe3+. The hydroxyl radicals were scavenged by the Abferritin protein in vitro and the ectopic expression of Abferritin could increase the survival ratios of E. coli cells under the oxidative stress. [Conclusion] Abferritin was strongly up regulated under oxidative stress and the Abferritin protein exhibited ferroxidase and anti-oxidative activity.
