Open AccessThe construction of a reporter system and use for the investigation of Clostridium perfringens gene expression
Author(s) -
Bullifent Helen L.,
Moir Anne,
Titball Richard W.
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07761.x
Subject(s) - chloramphenicol acetyltransferase , clostridium perfringens , open reading frame , shuttle vector , reporter gene , biology , acetyltransferase , gene , plasmid , gene expression , microbiology and biotechnology , bioreporter , genetics , vector (molecular biology) , recombinant dna , acetylation , bacteria , peptide sequence
A reporter system was constructed to enable the study of gene expression in Clostridium perfringens . The system was based on plasmid shuttle vector pJIR410, which contained the C. perfringens erythromycin resistance gene. The vector was modified by the introduction of a DNA fragment comprising the open reading frame of the C. perfringens chloramphenicol acetyltransferase gene and flanking transcriptional terminators. The presence of a unique restriction site, engineered into the extreme 5′ end of the open reading frame enabled a promoter region to be inserted to form an in‐frame transcriptional fusion with catP . The system was tested by inserting the promoter region of the alpha‐toxin gene of C. perfringens . The production of chloramphenicol acetyltransferase in C. perfringens was monitored during growth and the pattern of expression was shown to reflect levels of plc mRNA and alpha‐toxin in the parent strain.