Characterization of a Novel Oxidative Stress Responsive Transcription Regulator in Mycobacterium bovis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains, Primers and Media
2.2. Protein Expression and Purification
2.3. Real-Time Fluorescence Quantitative PCR Assay
2.4. Growth Curves Assays
2.5. DNA Substrate Preparation and Electrophoretic Mobility Shift Assay (EMSA)
2.6. ChIP Assay
2.7. DNA Footprinting Assay
2.8. β-Galactosidase Activity Assay
2.9. Determination of Intracellular ROS Content
2.10. Transcriptomic Analysis
3. Results
3.1. aotM Gene Is Required for Mycobacterium bovis to Resist Oxidative Stress
3.2. AotM Specifically Binds to Its Own Promoter Region
3.3. AotM Recognizes a 14-bp Palindrome Sequence Motif
3.4. AotM Affects the Expression of Genes Involved in Antioxidation in Mycobacterium bovis
3.5. AotM Negatively Regulates the Expression of the Adjacent Genes
3.6. Overexpression of BCG_3892c Leads to Increased Intracellular ROS Level and Reduced Growth of M. bovis
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Jiang, Q.; Hu, R.; Liu, F.; Huang, F.; Zhang, L.; Zhang, H. Characterization of a Novel Oxidative Stress Responsive Transcription Regulator in Mycobacterium bovis. Biomedicines 2024, 12, 1872. https://doi.org/10.3390/biomedicines12081872
Jiang Q, Hu R, Liu F, Huang F, Zhang L, Zhang H. Characterization of a Novel Oxidative Stress Responsive Transcription Regulator in Mycobacterium bovis. Biomedicines. 2024; 12(8):1872. https://doi.org/10.3390/biomedicines12081872
Chicago/Turabian StyleJiang, Qiang, Rong Hu, Feng Liu, Feng Huang, Lei Zhang, and Hua Zhang. 2024. "Characterization of a Novel Oxidative Stress Responsive Transcription Regulator in Mycobacterium bovis" Biomedicines 12, no. 8: 1872. https://doi.org/10.3390/biomedicines12081872