Creation of an isogenic P1-deficient mutant of Haemophilus influenzae biogroup aegyptius.
Haemophilus influenzae biogroup aegyptius, the causative agent of Brazilian purpuric fever (BPF), expresses a heat-modifiable 48 kDa outer membrane protein, P1, which is conserved in most Brazilian case-clone isolates. To study the role of P1 in pathogenesis of BPF we constructed via homologous recombination an isogenic P1-deficient mutant of H. influenzae biogroup aegyptius. The procedure involved a modification of Hererot's method for development of competence. Modifications included variations in the growth conditions, use of cAMP, specific characteristics of the donor DNA, and antibiotic selection. P1-deficient mutants were confirmed by SDS-PAGE, loss of reactivity with a specific monoclonal antibody on Western blot, restriction analysis and Southern blot. Our results establish the first successful transformation of homologous DNA into H. influenzae biogroup aegyptius.
Segada
L M
LM
Department of Microbiology, State University of New York at Buffalo, New York, USA.
Lesse
A J
AJ
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli; D017353:Gene Deletion; D005798:Genes, Bacterial; D015698:Genomic Library; D006193:Haemophilus influenzae / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D013489:Suppression, Genetic
D001425:Bacterial Outer Membrane Proteins; C067559:outer membrane protein P1, Haemophilus influenzae type b