Isolation of RNA Polymerase from Clostridium difficile and Characterization of Glutamate Dehydrogenase and rRNA Gene Promoters In Vitro and In Vivo
| Autor: | Abraham L. Sonenshein, Nagraj Mani, Bruno Dupuy |
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| Přispěvatelé: | Department of Molecular Biology and Microbiology [Tufts, Boston], Tufts University School of Medicine [Boston], Génétique Moléculaire Bactérienne, Institut Pasteur [Paris] (IP), This work was supported by research grants to A.L.S. (grants AI057637 and GM042219) and a project grant to the Tufts Gastroenterology Research and Secretory Processes Center (grant DK034928) from the U.S. Public Health Service, by a postdoctoral fellowship to N.M. from the Charles A. King Trust, and by research funds provided to B.D. by the Institut Pasteur, Paris, France., We thank L. Barosso and T. Wilkins for providing the cloned gdh promoter region, the Sanger Centre for making the C. difficile genome sequence available prior to publication, and B. Belitsky, K. Matsuno, and J. Singh for helpful discussions., Institut Pasteur [Paris] |
| Rok vydání: | 2006 |
| Předmět: |
Transcription
Genetic MESH: Clostridium difficile/genetics [SDV]Life Sciences [q-bio] Molecular Sequence Data MESH: Base Sequence Biology Microbiology 03 medical and health sciences chemistry.chemical_compound Glutamate Dehydrogenase MESH: Genes rRNA Transcription (biology) MESH: DNA-Directed RNA Polymerases/metabolism RNA polymerase Gene expression Transcriptional regulation Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: Clostridium difficile/enzymology Promoter Regions Genetic Molecular Biology Gene MESH: Transcription Genetic 030304 developmental biology Molecular Biology of Pathogens Regulation of gene expression MESH: Gene Expression Regulation Bacterial 0303 health sciences MESH: Humans MESH: Molecular Sequence Data Base Sequence Clostridioides difficile 030306 microbiology MESH: Glutamate Dehydrogenase/metabolism Genes rRNA Promoter DNA-Directed RNA Polymerases Gene Expression Regulation Bacterial Clostridium difficile Molecular biology MESH: DNA-Directed RNA Polymerases/isolation & purification MESH: Glutamate Dehydrogenase/genetics 3. Good health chemistry MESH: DNA-Directed RNA Polymerases/genetics MESH: Promoter Regions Genetic*/physiology |
| Zdroj: | Journal of Bacteriology Journal of Bacteriology, 2005, 188 (1), pp.96-102. ⟨10.1128/JB.188.1.96-102.2006⟩ Journal of Bacteriology, American Society for Microbiology, 2005, 188 (1), pp.96-102. ⟨10.1128/JB.188.1.96-102.2006⟩ |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.188.1.96-102.2006 |
| Popis: | Clostridium difficile is the primary causative agent of antibiotic-associated diarrheal disease. To facilitate molecular genetic analysis of gene expression in this organism, methods were developed to study transcriptional regulation in vitro and in vivo. That is, C. difficile RNA polymerase was partially purified and shown to bind to and initiate transcription in vitro from bona fide C. difficile promoters for rRNA and glutamate dehydrogenase genes. In addition, primer extension analyses and a β-glucuronidase reporter system were used to quantitate transcription from these promoters in vivo. With these tools in hand, it is now possible to characterize the behavior of any C. difficile gene in vivo and to study the regulation of its expression in detail. |
| Databáze: | OpenAIRE |
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