Hl. Bullifent et al., THE CONSTRUCTION OF A REPORTER SYSTEM AND USE FOR THE INVESTIGATION OF CLOSTRIDIUM-PERFRINGENS GENE-EXPRESSION, FEMS microbiology letters, 131(1), 1995, pp. 99-105
A reporter system was constructed to enable the study of gene expressi
on in Clostridium perfringens. The system was based on plasmid shuttle
vector pJIR410, which contained the C. perfringens erythromycin resis
tance gene. The vector was modified by the introduction of a DNA fragm
ent comprising the open reading frame of the C. perfringens chloramphe
nicol acetyltransferase gene and flanking transcriptional terminators.
The presence of a unique restriction site, engineered into the extrem
e 5' end of the open reading frame enabled a promoter region to be ins
erted to form an in-frame transcriptional fusion with catP. The system
was tested by inserting the promoter region of the alpha-toxin gene o
f C. perfringens. The production of chloramphenicol acetyltransferase
in C. perfringens was monitored during growth and the pattern of expre
ssion was shown to reflect levels of pie mRNA and alpha-toxin in the p
arent strain.