Detection of the outer membrane lipoprotein I and its gene in fluorescent and non-fluorescent pseudomonads: implications for taxonomy and diagnosis.
The open reading frame of the OprI lipoprotein gene from Pseudomonas aeruginosa was amplified by polymerase chain reaction (PCR) starting from purified DNA or colony lysates. A fragment of the expected size (249 bp) was detected in all P. aeruginosa strains from various clinical and geographical origins. The gene could only be amplified in pseudomonads of rRNA group I which are considered to be the authentic genus Pseudomonas. Digestions with HaeIII, PvuII and SphI of the amplified fragments demonstrated a sequence variation in the oprI gene. Colony, dot and Western blots with two monoclonal antibodies (mAbs) against the lipoprotein I confirmed our PCR results. These findings open interesting perspectives for the molecular taxonomy of the genus Pseudomonas and the development of diagnostic tools.
De Vos
D
D
Vrije Universiteit Brussel, Belgium.
Lim
A
A
De Vos
P
P
Sarniguet
A
A
Kersters
K
K
Cornelis
P
P
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005453:Fluorescence; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics; D006801:Humans; D008074:Lipoproteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011549:Pseudomonas / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D011552:Pseudomonas Infections / Q000175:diagnosis; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000145:classification / Q000235:genetics; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008074:Lipoproteins; C060507:outer membrane lipoprotein I, Bacteria