title: Identification and characterization of an anti-oxidative stress-associated mutant of Aspergillus fumigatus transformed by Agrobacterium tumefaciens.
abstract: Aspergillus fumigatus is one of the most common opportunistic pathogenic fungi, surviving in various environmental conditions. Maintenance of the redox homeostasis of the fungus relies upon the well‑organized regulation between reactive oxygen species generated by immune cells or its own organelles, and the activated anti‑oxidative stress mechanism. To investigate such a mechanism, the present study obtained a number of randomly‑inserted mutants of A. fumigatus, mediated by Agrobacterium tumefaciens. In addition, a high throughput hydrogen peroxide screening system was established to examine ~1,000 mutants. A total of 100 mutants exhibited changes in hydrogen peroxide sensitivity, among which a significant increase in sensitivity was observed in the AFM2658 mutant. Further investigations of the mutant were also performed, in which the sequence of this mutant was characterized using thermal asymmetric interlaced‑polymerase chain reaction. This revealed that the insertion site was located on chromosome 2 afu1_92, and the 96 bp sequence was knocked out, which partially comprised a sequence localized between the integral membrane protein coding region and the helix‑loop‑helix transcription factor coding region. A decrease in the levels of anti‑oxidative stress‑associated mRNAs were observed, and an increase in reactive oxygen species were detected using fluorescence. The results of the present study demonstrated that this sequence may have a protective role in A. fumigatus in the presence of oxidative stress.
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Zhongqi
authors: Z
authors: Fan
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Huimei
authors: H
authors: Yu
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Qi
authors: Q
authors: Guo
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Dan
authors: D
authors: He
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Baiji
authors: B
authors: Xue
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Xiangli
authors: X
authors: Xie
authors: Medical Mycology Research Center, Chiba University, Chiba 260‑8673, Japan.
authors: Koji
authors: K
authors: Yokoyama
authors: Department of Pathogenobiology, Jilin University Mycology Research Center, Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
authors: Li
authors: L
authors: Wang
mesh_terms: D016960:Agrobacterium tumefaciens / Q000187:drug effects / Q000378:metabolism*; D000935:Antifungal Agents / Q000494:pharmacology; D001232:Aspergillus fumigatus / Q000235:genetics* / Q000254:growth & development; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics; D005452:Fluoresceins / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D016254:Mutagenesis, Insertional / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics*; D013696:Temperature; D014158:Transcription, Genetic / Q000187:drug effects; D014170:Transformation, Genetic / Q000187:drug effects
chemical_list: D000935:Antifungal Agents; D004269:DNA, Bacterial; D005452:Fluoresceins; C036483:T-DNA; C029569:diacetyldichlorofluorescein; D006861:Hydrogen Peroxide