Please use this identifier to cite or link to this item: https://doi.org/10.1128/JB.184.7.1932-1939.2002
Title: Identification and characterization of CAMP cohemolysin as a potential virulence factor of Riemerella anatipestifer
Authors: Crasta, K.C.
Chua, K.-L.
Subramaniam, S. 
Frey, J.
Loh, H.
Tan, H.-M.
Issue Date: 2002
Citation: Crasta, K.C., Chua, K.-L., Subramaniam, S., Frey, J., Loh, H., Tan, H.-M. (2002). Identification and characterization of CAMP cohemolysin as a potential virulence factor of Riemerella anatipestifer. Journal of Bacteriology 184 (7) : 1932-1939. ScholarBank@NUS Repository. https://doi.org/10.1128/JB.184.7.1932-1939.2002
Abstract: Riemerella anatipestifer is responsible for exudative septicemia in ducks. The genetic determinant of the CAMP cohemolysin, cam, from a strain of R. anatipestifer was cloned and expressed in Escherichia coli. Chromosomal DNA from serotype 19 strain 30/90 was used to construct a gene library in pBluescript II SK(-) vector in E. coli XL-1-Blue strain. The clones containing recombinant plasmids were screened for the CAMP reaction with Staphylococcus aureus. Those that showed cohemolysis were chosen for further analysis by sequencing. One of these clones, JFRA8, was subcloned to identify the smallest possible DNA fragment containing the CAMP cohemolysin determinant, which was located on a 3,566-bp BamHI-BstXI fragment which specified a 1,026-bp open reading frame. Clones containing recombinant plasmids carrying cam obtained by PCR cloning into E. coli M15 strain secreted an active CAMP cohemolysin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses confirmed that the recombinant strain expressed a protein with a molecular mass of 37 kDa and that strains from serotypes 1, 2, 3, 5, 6, and 19 expressed the cohemolysin. The deduced amino acid sequence showed high homology to those of O-sialoglycoprotein endopeptidases. Hydrolysis of radioiodinated glycophorin A confirmed that Cam is a sialoglycoprotease.
Source Title: Journal of Bacteriology
URI: http://scholarbank.nus.edu.sg/handle/10635/112928
ISSN: 00219193
DOI: 10.1128/JB.184.7.1932-1939.2002
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