TY - JOUR
T1 - Development of a genetic manipulation system for Haemophilus parasuis
AU - Bigas, Anna
AU - Garrido, M. Elena
AU - Pérez De Rozas, Ana M.
AU - Badiola, Ignacio
AU - Barbé, Jordi
AU - Llagostera, Montserrat
PY - 2005/2/25
Y1 - 2005/2/25
N2 - 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. © 2004 Elsevier B.V. All rights reserved.
AB - 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. © 2004 Elsevier B.V. All rights reserved.
KW - DNA transformation
KW - Genetic manipulation
KW - Haemophilus parasuis
KW - Knockout mutants
U2 - 10.1016/j.vetmic.2004.10.015
DO - 10.1016/j.vetmic.2004.10.015
M3 - Article
SN - 0378-1135
VL - 105
SP - 223
EP - 228
JO - Veterinary Microbiology
JF - Veterinary Microbiology
IS - 3-4
ER -