title: Sequencing and diversity analyses reveal extensive similarities between some epsilon-toxin-encoding plasmids and the pCPF5603 Clostridium perfringens enterotoxin plasmid.
abstract: Clostridium perfringens type B and D isolates produce epsilon-toxin, the third most potent clostridial toxin. The epsilon-toxin gene (etx) is plasmid borne in type D isolates, but etx genetics have been poorly studied in type B isolates. This study reports the first sequencing of any etx plasmid, i.e., pCP8533etx, from type B strain NCTC8533. This etx plasmid is 64.7 kb, carries tcp conjugative transfer genes, and encodes additional potential virulence factors including beta2-toxin, sortase, and collagen adhesin but not beta-toxin. Interestingly, nearly 80% of pCP8533etx open reading frames (ORFs) are also present on pCPF5603, an enterotoxin-encoding plasmid from type A isolate F5603. Pulsed-field gel electrophoresis and overlapping PCR indicated that a pCP8533etx-like etx plasmid is also present in most, if not all, other type B isolates and some beta2-toxin-positive, cpe-negative type D isolates, while other type D isolates carry different etx plasmids. Sequences upstream of the etx gene vary between type B isolates and some type D isolates that do not carry a pCP8533etx-like etx plasmid. However, nearly all type B and D isolates have an etx locus with an upstream IS1151, and those etx loci typically reside near a dcm ORF. These results suggest that pCPF5603 and pCP8533etx evolved from insertion of mobile genetic elements carrying enterotoxin or etx genes, respectively, onto a common progenitor plasmid.
authors.affiliation: Department of Microbiology, Wakayama Medical University School of Medicine, Wakayama, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Jihong
authors.initials: J
authors.lastname: Li
authors.forename: Sameera
authors.initials: S
authors.lastname: Sayeed
authors.forename: Shigeru
authors.initials: S
authors.lastname: Akimoto
authors.forename: Bruce A
authors.initials: BA
authors.lastname: McClane
mesh_terms: D015139:Blotting, Southern; D003016:Clostridium perfringens / Q000235:genetics* / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D004768:Enterotoxins / Q000378:metabolism*; D014644:Genetic Variation; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D004768:Enterotoxins
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