This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mani, N.
Right arrow Articles by Sonenshein, A. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mani, N.
Right arrow Articles by Sonenshein, A. L.

 Previous Article  |  Next Article 

Journal of Bacteriology, January 2006, p. 96-102, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.96-102.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Isolation of RNA Polymerase from Clostridium difficile and Characterization of Glutamate Dehydrogenase and rRNA Gene Promoters In Vitro and In Vivo

Nagraj Mani,1,{dagger} Bruno Dupuy,2 and Abraham L. Sonenshein1*

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111,1 Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, Paris, France2

Received 25 August 2005/ Accepted 7 October 2005

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 ß-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.

* Corresponding author. Mailing address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111. Phone: (617) 636-6761. Fax: (617) 636-0337. E-mail: linc.sonenshein{at}tufts.edu.

{dagger} Present address: Vertex Pharmaceuticals, Inc., 130 Waverly St., Cambridge, MA 02139.

Journal of Bacteriology, January 2006, p. 96-102, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.96-102.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»