Development of a genetic manipulation system for Haemophilus parasuis

Haemophilus parasuis is a member of the family Pasteurellaceae and an important respiratory-tract pathogen of swine, which is the etiological agent of Glässer's disease. Because no genetic manipulation system is available for H. parasuis so far, in vivo studies about the role of its genes invol...

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Bibliographic Details
Published in:Veterinary microbiology 2005-02, Vol.105 (3), p.223-228
Main Authors: Bigas, Anna, Garrido, M. Elena, Rozas, Ana M. Pérez de, Badiola, Ignacio, Barbé, Jordi, Llagostera, Montserrat
Format: Article
Language:English
Subjects:
Animals
arthritis
bacterial infections
Bacteriology
Base Sequence
Biological and medical sciences
cyclic AMP
Cyclic AMP - genetics
Cyclic AMP - physiology
DNA transformation
DNA, Bacterial - analysis
Fundamental and applied biological sciences. Psychology
genetic engineering
Genetic manipulation
genetic techniques and protocols
genetic transformation
Glasser's disease
Haemophilus parasuis
Haemophilus parasuis - genetics
Haemophilus parasuis - pathogenicity
Knockout mutants
Microbiology
Miscellaneous
Mitochondrial Proteins - genetics
Molecular Sequence Data
Mutation
nucleotide sequences
Pasteurellaceae
pathogenesis
poliserositis
respiratory tract diseases
Ribosomal Proteins - genetics
Swine
swine diseases
thymidylate synthase
Thymidylate Synthase - genetics
Transformation, Bacterial
Virulence - genetics
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Summary:Haemophilus parasuis is a member of the family Pasteurellaceae and an important respiratory-tract pathogen of swine, which is the etiological agent of Glässer's disease. Because no genetic manipulation system is available for H. parasuis so far, in vivo studies about the role of its genes involved in virulence are unfeasible. Here we demonstrate that H. parasuis has a cyclic AMP (cAMP)-dependent natural transformation system that enables the uptake of DNA in which the ACCGAACTC sequence signal must be present. After improving DNA transformation parameters, such as cAMP and DNA concentration and exposition time of the exogenous DNA, a knockout mutant of H. parasuis defective in the thy gene, encoding the thymidylate synthase enzyme, has been constructed. Data presented in this work open the possibility for the functional analysis of genes involved in the infectious process of this animal pathogen.
ISSN:0378-1135
1873-2542
DOI:10.1016/j.vetmic.2004.10.015