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PMID: 2659534 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and expression of a cohemolysin, the CAMP factor of Actinobacillus pleuropneumoniae.

Infection and immunity ·Vol. 57 ·No. 7 ·1989-07-00 ·Pages 2050-6

Frey J, Perrin J, Nicolet J

Abstract

The genetic determinant of the cohemolysin which is responsible for the CAMP phenomenon, a cohemolysis, of Actinobacillus pleuropneumoniae was cloned in Escherichia coli. Total DNA from the A. pleuropneumoniae serotype 1 type strain 4074 was used to construct a gene library in plasmid pUC18 in E. coli JM83. A total of 10,500 clones containing recombinant plasmids have been screened for hemolysis on blood plates. Fifty-five clones which showed a weak hemolytic response after 24 to 48 h of incubation were screened for the CAMP reaction with Staphylococcus aureus. This led to the identification of one clone which showed a positive CAMP reaction. Immunoblot analysis revealed that the recombinant strain expressed a protein with a molecular mass of 27,000 daltons, similar in size to the CAMP protein of the group B streptococci. Rabbit antibodies against the CAMP+ clone neutralized the CAMP reaction mediated by the E. coli strain containing the cloned CAMP gene as well as that of A. pleuropneumoniae. Antibodies raised against the cloned CAMP cohemolysin cross-reacted with Streptococcus agalactiae protein B. We designate the 27,000-dalton molecule CAMP factor protein and name its corresponding gene cfp.

MeSH Terms
Actinobacillus/classification,genetics Bacterial Proteins/biosynthesis,genetics,isolation & purification Blotting, Western Cloning, Molecular/methods Escherichia coli/genetics Hemolysin Proteins/biosynthesis,genetics,isolation & purification Serotyping Streptococcus agalactiae/isolation & purification
Chemicals
Bacterial Proteins CAMP protein, Streptococcus Hemolysin Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Frey J
Institute for Veterinary Bacteriology, University of Berne, Switzerland.
Perrin J
Nicolet J
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.57.7.2050-2056.1989
Published
1989-07-00
Pages
2050-6
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC313840
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.57.7.2050-2056.1989 PMC Full Text View journal details
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