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Journal of Bacteriology, April 2006, p. 2568-2577, Vol. 188, No. 7
0021-9193/06/$08.00+0     doi:10.1128/JB.188.7.2568-2577.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genomic Organization and Molecular Characterization of Clostridium difficile Bacteriophage {Phi}CD119

Revathi Govind, Joe A. Fralick, and Rial D. Rolfe*

Department of Microbiology and Immunology, Texas Tech University Health Sciences Center, Lubbock, Texas 79430

Received 10 October 2005/ Accepted 13 January 2006

In this study, we have isolated a temperate phage ({Phi}CD119) from a pathogenic Clostridium difficile strain and sequenced and annotated its genome. This virus has an icosahedral capsid and a contractile tail covered by a sheath and contains a double-stranded DNA genome. It belongs to the Myoviridae family of the tailed phages and the order Caudovirales. The genome was circularly permuted, with no physical ends detected by sequencing or restriction enzyme digestion analysis, and lacked a cos site. The DNA sequence of this phage consists of 53,325 bp, which carries 79 putative open reading frames (ORFs). A function could be assigned to 23 putative gene products, based upon bioinformatic analyses. The {Phi}CD119 genome is organized in a modular format, which includes modules for lysogeny, DNA replication, DNA packaging, structural proteins, and host cell lysis. The {Phi}CD119 attachment site attP lies in a noncoding region close to the putative integrase (int) gene. We have identified the phage integration site on the C. difficile chromosome (attB) located in a noncoding region just upstream of gene gltP, which encodes a carrier protein for glutamate and aspartate. This genetic analysis represents the first complete DNA sequence and annotation of a C. difficile phage.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Health Sciences Center, Texas Tech University, School of Medicine, Lubbock, TX 79430. Phone: (806) 743-2905. Fax: (806) 743-2334. E-mail: rial.rolfe{at}ttuhsc.edu.

Journal of Bacteriology, April 2006, p. 2568-2577, Vol. 188, No. 7
0021-9193/06/$08.00+0     doi:10.1128/JB.188.7.2568-2577.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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