1887

Abstract

Immunological screening of a cosmid library led to the identification of clones producing an 18 kDa outer-membrane protein. This protein reacted in Western blots with a polyclonal antiserum against outer-membrane proteins of and with a monoclonal antibody (MA1-6) specific for OprL, the peptidoglycan-associated outer-membrane lipoprotein (PAL). Sequencing of pOML7, a subclone expressing revealed an ORF of 504 bp encoding a polypeptide with a typical lipoprotein signal recognition sequence. Another ORF was found upstream of with homology to the ToIB protein of and Downstream of a second ORF, of 321 bp, was found (), encoding a protein with a signal peptide and with no homology with proteins of known biological function. After the stop codon of a rho-independent terminator sequence was detected which is part of the PA01 insertion element IS222. OprL showed homologies with all known PALs from Gram-negative bacteria, especially in the C-terminal part. mAb MA1-6 reacted with cells in immunofluorescence, and with cells expressing which had an abnormal, elongated morphology, an indication that production of the protein perturbed the division process.

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1997-05-01
2021-10-22
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