1887

Abstract

A Tn mutant library was generated to identify genes involved in the biogenesis of fimbriae. A fimbria-deficient mutant was isolated by negative selection using an immunomagnetic separation technique with specific anti-fimbriae polyclonal antibodies (pAbs). The transposon was inserted in an ORF, called , which encoded a protein of unknown function. The transposon prevented the transcription of as well as two genes located downstream, which are designated and and which form the operon. Sequence analyses of CspA and CspB revealed that both proteins possessed the classic cell-wall-anchoring motif (LPXTG) of Gram-positive bacterial surface proteins. Recombinant CspA (rCspA) and CspB (rCspB) proteins were generated in and used to produce pAbs. Immunolocalization experiments showed that anti-rCspB, but not anti-rCspA antibodies specifically recognized fimbriae. Our results suggested that the operon encoded predicted cell-surface proteins, one of which, CspB, was associated with the fimbriae.

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2004-01-01
2020-10-19
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