Cloning and analysis of the gene for the major outer membrane lipoprotein from Pseudomonas aeruginosa.
The gene for the Pseudomonas aeruginosa outer membrane lipoprotein I was isolated from a genomic library in the phage lambda EMBL3 vector and subsequently subcloned in the low copy-number, wide host-range plasmid vector, pKT240. The cloned gene was highly expressed, resulting in the production of a low molecular-weight protein (8 kD) that was found to be associated with the outer membrane. Sequence analysis showed an open reading frame of 83 amino acids with a putative N-terminal hydrophobic signal peptide of 19 residues immediately followed by the lipoprotein consensus sequence, GLY-CYS-SER-SER (residues 19-22). The predicted amino acid composition of the mature polypeptide and that of the purified lipoprotein I of P. aeruginosa (Mizuno and Kageyama, 1979) were identical. In contrast with other Gram-negative outer membrane lipoproteins, conformation predictions suggested that the mature protein was a single alpha helix.
Cornelis
P
P
Laboratoire de Microbiologie, CNRS, Université Louis Pasteur, Strasbourg, France.
Bouia
A
A
Belarbi
A
A
Guyonvarch
A
A
Kammerer
B
B
Hannaert
V
V
Hubert
J C
JC
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D000918:Antibody Specificity; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins; D004755:Enterobacteriaceae / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000276:immunology; D012329:RNA, Bacterial / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C031908:H-NS protein, bacteria; D012329:RNA, Bacterial; D011994:Recombinant Proteins