Regulated expression of Clostridium perfringens enterotoxin in naturally cpe-negative type A, B, and C isolates of C. perfringens

Abstract

Clostridium perfringens enterotoxin (CPE), the virulence factor responsible for symptoms associated with C. perfringens type A food poisoning, is produced by enterotoxigenic C. perfringens type A isolates when these bacteria sporulate in the gastrointestinal tract. Less than 5% of the global C. perfringens population apparently carries the cpe gene. To assess the distribution of cpe-regulatory factors, we investigated whether the cpe gene of a C. perfringens food poisoning isolate can be expressed and properly regulated (i.e., expressed in a sporulation-associated manner) when transformed into naturally cpe-negative C. perfringens isolates. Sporulation-associated CPE expression was observed when low-copy-number plasmids carrying either a 5.7-kb DNA insert, containing the cpe open reading frame plus >1 kb each of upstream and downstream flanking sequences from C. perfringens food poisoning isolate NCTC 8239, or a 1.6-kb insert, containing only the cpe open reading frame of NCTC 8239, were electroporated into cpe-negative C. perfringens type A, B, and C isolates. Northern (RNA) blot analysis demonstrated that the sizes of the cpe message in the transformants and the naturally enterotoxigenic C. perfringens NCTC 8239 were similar and that this message was detectable only in sporulating cultures of the transformants or NCTC 8239. These studies strongly suggest that many, if not all, cpe-negative C. perfringens isolates (including type B isolates, which are not known to naturally express CPE) produce a factor(s) involved in normal (i.e., sporulation-associated) transcriptional regulation of CPE expression by C. perfringens food poisoning isolates. These findings are consistent with this CPE-regulatory factor(s) also regulating the expression of other genes in C. perfringens.