THE CONSTRUCTION OF A REPORTER SYSTEM AND USE FOR THE INVESTIGATION OF CLOSTRIDIUM-PERFRINGENS GENE-EXPRESSION

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.