Open AccessCloning, nucleotide sequence and expression of a hemolysin gene of Clostridium septicum
Author(s) -
Imagawa Tadashi,
Dohi Yoshitane,
Higashi Yasushi
Publication year - 1994
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.1574-6968.1994.tb06781.x
Subject(s) - hemolysin , clostridium septicum , biology , nucleic acid sequence , plasmid , microbiology and biotechnology , escherichia coli , gene , open reading frame , insert (composites) , genomic library , genomic dna , genetics , peptide sequence , virulence , mechanical engineering , engineering
A genomic library of Clostridium septicum NCTC547 strain was made in Escherichia coli by means of λgt10. The DNA insert of a hemolysin‐positive (Hly + ) λ‐clone was transferred into pUC19. The resulting plasmid, pCS21, confers a Hly + phenotype on E. coli . Crude lysates of E. coli (pCS21) possessed a strong lytic activity on human erythrocytes and also a lethal effect on mice, characteristic of an α toxin. Nucleotide sequence analysis revealed that the insert DNA (5.2 kb) in pCS21 included at least one open reading frame of 1380 bp. The coding frame for hemolysin was predicted to be 1329 bp in size and to encode a protein of 49.8 kDa. It coincided with the molecular mass (48 kDa) of the α toxin secreted by C. septicum . Taken together, the data indicated that plasmid pCS21 indeed encoded an α toxin gene of C. septicum .