Bioluminescence (lux) expression in the anaerobe Clostridium perfringens

To determine whether bacterial luciferase is expressed in the anaerobe Clostridium perfringens to produce an oxygen-requiring bioluminescence reaction, a suitable plasmid vector possessing the luxA and luxB genes of Vibrio fischeri was constructed and introduced into C. perfringens cells. luxAB were placed under the transcriptional control of the C. perfringens alpha-toxin gene promoter region. Suitable ribosome binding sites were introduced upstream of both genes. Bioluminescence was strongly expressed in C. perfringens transformants. Comparisons of in vivo and in vitro bioluminescence measurements demonstrated that in vivo data constituted a quantitative measure of gene expression. This is the first study to show that luxA and luxB genes can be expressed in an anaerobic bacterium and that bioluminescence can be used as a quantitative reporter system in future in vivo studies of gene expression in C. perfringens.