Abstract
Transcriptomics enables us to elucidate comprehensive gene expression profiles in given experimental conditions. Global regulators, which include transcriptional regulators and two-component regulatory systems, have evolved in a variety of bacterial systems. They play important roles in bacterial fitness and pathogenesis by regulating target gene expression. Advanced next-generation RNA sequencing technology (RNA-seq) provides a powerful and effective tool to analyze the transcriptome of bacterial cells. In this chapter, we provide a detailed procedure for the investigation of gene expression profiles and identification of target genes, regulons, and/or pathways that are mediated by a regulator. This procedure is done using RNA-seq analysis, which involves RNA purification, mRNA enrichment, decontamination, RNA-seq data analysis, and quantitative real-time reverse transcription PCR.
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Acknowledgments
We thank Dr. Juan E Abrahante Llorens at the University of Minnesota Informatics Institute (UMII) and Lisa Fazzino for the assistance with data analysis. This work was supported by grant AI057451 from the National Institute of Allergy and Infectious Disease and partially supported by grant MIN-63-082 and MIN-63-113 from CVM research Office UMN Ag Experimental Station General Agricultural Research Funds.
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Lei, T., Yang, J., Becker, A., Ji, Y. (2020). Identification of Target Genes Mediated by Two-Component Regulators of Staphylococcus aureus Using RNA-seq Technology. In: Ji, Y. (eds) Methicillin-Resistant Staphylococcus Aureus (MRSA) Protocols. Methods in Molecular Biology, vol 2069. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9849-4_10
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DOI: https://doi.org/10.1007/978-1-4939-9849-4_10
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