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PMID: 2536355 Published · ppublish English Journal Article

Molecular cloning and nucleotide sequence of the alpha-toxin (phospholipase C) of Clostridium perfringens.

Infection and immunity ·Vol. 57 ·No. 2 ·1989-02-00 ·Pages 367-76

Titball RW, Hunter SE, Martin KL, Morris BC, Shuttleworth AD, Rubidge T, Anderson DW, Kelly DC

Abstract

A fragment of DNA containing the gene coding for the phospholipase C (alpha-toxin) of Clostridium perfringens was cloned into Escherichia coli. The cloned DNA appeared to code only for the alpha-toxin and contained both the coding region and its associated gene promoter. The nucleotide sequence of the cloned DNA was determined, and an open reading frame was identified which encoded a protein with a molecular weight of 42,528. By comparison of the gene sequence with the N-terminal amino acid sequence of the protein, a 28-amino-acid signal sequence was identified. The gene promoter showed considerable homology with the E. coli sigma 55 consensus promoter sequences, and this may explain why the gene was expressed by E. coli. The cloned gene product appeared to be virtually identical to the native protein. A 77-amino-acid stretch that was close to the N terminus of the alpha-toxin showed considerable homology with similarly located regions of the Bacillus cereus phosphatidylcholine, preferring phospholipase C and weaker homology with the phospholipase C from Pseudomonas aeruginosa.

MeSH Terms
Amino Acid Sequence Base Sequence Chemical Phenomena Chemistry, Physical Cloning, Molecular Clostridium perfringens/enzymology,genetics DNA, Bacterial/isolation & purification DNA, Recombinant/isolation & purification Molecular Sequence Data Nucleotide Mapping Plasmids Sequence Homology, Nucleic Acid Type C Phospholipases/genetics,isolation & purification
Chemicals
DNA, Bacterial DNA, Recombinant Type C Phospholipases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Titball R W
Chemical Defence Establishment, Salisbury, Wiltshire, United Kingdom.
Hunter S E
Martin K L
Morris B C
Shuttleworth A D
Rubidge T
Anderson D W
Kelly D C
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.57.2.367-376.1989
Published
1989-02-00
Pages
367-76
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC313106
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.57.2.367-376.1989 PMC Full Text View journal details
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