Identification of an alkaline ceramidase gene from Dermatophilus congolensis

A random amplified polymorphic DNA (RAPD) procedure was used to identify a specific 0.6 kb DNA fragment unique to Dermatophilus congolensis. This 0.6 kb fragment was evaluated as a specific DNA probe and used to design oligonucleotide primers for polymerase chain reaction (PCR) amplification. The nu...

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Bibliographic Details
Published in:Veterinary microbiology 2004-03, Vol.99 (1), p.67-74
Main Authors: Garcı́a-Sánchez, Alfredo, Cerrato, Rosario, Larrasa, José, Ambrose, Nicholas C, Parra, Alberto, Alonso, Juan M, Hermoso-de-Mendoza, Miguel, Rey, Joaquı́n M, Hermoso-de-Mendoza, Javier
Format: Article
Language:English
Subjects:
Actinobacteria - enzymology
Actinobacteria - genetics
Actinomycetales Infections - microbiology
Actinomycetales Infections - veterinary
agc gene
Alkaline ceramidase
Amidohydrolases - chemistry
Amidohydrolases - genetics
Amino Acid Sequence
Animals
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Base Sequence
Biological and medical sciences
Blotting, Southern
Ceramidases
ceramides
Dermatophilus congolensis
DNA
DNA primers
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Fundamental and applied biological sciences. Psychology
genes
Inverse PCR
Isoelectric Point
Microbiology
Molecular Sequence Data
Molecular Weight
open reading frames
Pseudomonas aeruginosa
Random Amplified Polymorphic DNA Technique
RAPD
Sequence Alignment
signal peptide
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Description
Summary:A random amplified polymorphic DNA (RAPD) procedure was used to identify a specific 0.6 kb DNA fragment unique to Dermatophilus congolensis. This 0.6 kb fragment was evaluated as a specific DNA probe and used to design oligonucleotide primers for polymerase chain reaction (PCR) amplification. The nucleotide sequences adjacent to this DNA fragment were determined by inverse PCR allowing the identification of a 4.1 kb sequence. Analysis of this revealed a complete open reading frame (ORF) with a high similarity to an alkaline ceramidase from Pseudomonas aeruginosa. The molecular weight of the enzyme derived from the predicted amino acid sequence is 74,662 Da, its p I is 9.81. The predicted N-terminal sequence of the enzyme contains a signal sequence indicating that the enzyme is exported by the bacterium. Since ceramides have important protective and cell regulatory roles in the epidermis we suggest that this ceramidase may have a role in the pathogenesis of dermatophilosis. It is the first completely sequenced gene described for D. congolensis.
ISSN:0378-1135
1873-2542
DOI:10.1016/j.vetmic.2003.10.028