Molecular cloning and characterisation of the genes for a non-fimbrial adhesin from Escherichia coli

Barbara A. Hales, Helen Beverley-Clarke, Nicky J. High, Klaus Jann, Rachel Perry, Janina Goldhar, Graham J. Boulnois*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

A non-fimbrial adhesin (NFA-1) from the uropathogenic Escherichia coli strain 827 responsible for agglutination of human erythrocytes was cloned using the cos 4 cosmid vector. A clone was isolated which promoted haemagglutination and showed the same biological properties as the adhesin produced by the wild type strain. Both express adhesin at 37 °C, but not 18 °C nor in the presence of 1% glucose. Adhesin purified from the clone formed high molecular weight aggregates which were resolved to the 21 K dalton subunit protein seen in the wild type strain on denaturation. Binding to human kidney cells by the clone and the wild type E. coli, from which the genes were cloned, were compared in an ELISA assay and shown to be the same. The genes for the adhesin were isolated on a 15.5 kilobase BamHI-EcoRI fragment which was subjected to γδ mutagenesis. The NFA-1 operon was localised to a 6.5kb region of this fragment.

Original languageEnglish
Pages (from-to)9-17
Number of pages9
JournalMicrobial Pathogenesis
Volume5
Issue number1
DOIs
StatePublished - Jul 1988

Funding

FundersFunder number
Bundes Ministerium fur Forschung and Technologie
Wellcome Trust
Medical Research Council

    Keywords

    • adhesin
    • cloning
    • haemagglutination
    • mutagenesis
    • non-fimbrial
    • protein purification

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