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Journal of Bacteriology, July 2006, p. 4942-4951, Vol. 188, No. 13
0021-9193/06/$08.00+0     doi:10.1128/JB.00298-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Functional Identification of Conjugation and Replication Regions of the Tetracycline Resistance Plasmid pCW3 from Clostridium perfringens

Trudi L. Bannam,{dagger} Wee Lin Teng,{dagger} Dieter Bulach, Dena Lyras, and Julian I. Rood*

Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics and Victorian Bioinformatics Consortium, Department of Microbiology, Monash University, Victoria 3800, Australia

Received 28 February 2006/ Accepted 16 April 2006

Clostridium perfringens causes fatal human infections, such as gas gangrene, as well as gastrointestinal diseases in both humans and animals. Detailed molecular analysis of the tetracycline resistance plasmid pCW3 from C. perfringens has shown that it represents the prototype of a unique family of conjugative antibiotic resistance and virulence plasmids. We have identified the pCW3 replication region by deletion and transposon mutagenesis and showed that the essential rep gene encoded a basic protein with no similarity to any known plasmid replication proteins. An 11-gene conjugation locus containing 5 genes that encoded putative proteins with similarity to proteins from the conjugative transposon Tn916 was identified, although the genes’ genetic arrangements were different. Functional genetic studies demonstrated that two of the genes in this transfer clostridial plasmid (tcp) locus, tcpF and tcpH, were essential for the conjugative transfer of pCW3, and comparative analysis confirmed that the tcp locus was not confined to pCW3. The conjugation region was present on all known conjugative plasmids from C. perfringens, including an enterotoxin plasmid and other toxin plasmids. These results have significant implications for plasmid evolution, as they provide evidence that a nonreplicating Tn916-like element can evolve to become the conjugation locus of replicating plasmids that carry major virulence genes or antibiotic resistance determinants.

* Corresponding author. Mailing address: Department of Microbiology, Monash University, Victoria 3800, Australia. Phone: (613) 9905-4825. Fax: (613) 9905-4811. E-mail: julian.rood{at}med.monash.edu.au.

{dagger} These authors contributed equally to this paper and are joint first authors.

Journal of Bacteriology, July 2006, p. 4942-4951, Vol. 188, No. 13
0021-9193/06/$08.00+0     doi:10.1128/JB.00298-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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