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research-article

Protein secretion in Pseudomonas aeruginosa: molecular cloning and characterization of the xcp-1 gene.

Laboratoire de Chimie Bactérienne, Centre National de la Recherche Scientifique, Marseilles, France.

ABSTRACT

Pleiotropic mutations (xcp) affecting the secretion of proteins in Pseudomonas aeruginosa have been previously characterized and mapped at three different loci on the chromosome. The xcp-1 gene, which is located at the 0-min region, was isolated from a genomic bank containing DNA from P. aeruginosa PAO1. The recombinant cosmid pLX25 complemented the xcp::Tn5-751 insertion mutation previously described. The xcp-1 gene was located on pLX25 by mapping the insertion point of transposon Tn5-751 and by deletion and subcloning analysis. The xcp-1 gene was expressed when transcription was initiated from a tac promoter, and a 26,000-dalton protein was identified in Escherichia coli minicells. The Xcp-1 protein was associated with the membrane fraction of E. coli. A 30-kilodalton outer membrane protein was also affected by the xcp::Tn5-751 mutation in P. aeruginosa. The possible correlation between Xcp-1 and this protein is discussed.

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