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Cloning and analysis of the gene for the major outer membrane lipoprotein from Pseudomonas aeruginosa
Author(s) -
Cornelis P.,
Bouia A.,
Belarbi A.,
Guyonvarch A.,
Kammerer B.,
Hannaert V.,
Hubert J. C.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb00187.x
Subject(s) - biology , bacterial outer membrane , peptide sequence , open reading frame , molecular cloning , amino acid , signal peptide , plasmid , pseudomonas aeruginosa , sequence analysis , gene , biochemistry , microbiology and biotechnology , cloning (programming) , cloning vector , genetics , escherichia coli , bacteria , computer science , programming language
Summary The gene for the Pseudomonas aeruginosa outer membrane lipoprotein I was isolated from a genomic library in the phage λ 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 (8kD) that was found to be associated with the outer membrane. Sequence anatysis 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.