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CLONING AND EXPRESSION A COLONOCYTE-SPECIFIC FIMBRIAL ADHESIN FROM AN ESCHERICHIA COLI STRAIN ISOLATED FROM AN INFANT WITH SEVERE DYSENTERY-LIKE DIARRHOEA

E.coli strain 469-3 (021 :NM) expresses mannose-resistant haemagglutination (MRHA) of human erythrocytes, adheres to cultured human epithelial (HEp-2) cells and to the brush border of human colonic (but not duodenal) enterocytes. The adhesin, an aggregate of a 14 kilodalton protein subunit, has now been identified by electron microscopy and consists of fine ∼2-nm diameter fibrils. The chromosomally located genetic determinants of the adhesin were isolated by cosmid cloning and expressed in E.coli K12. Several recombinant cosmids expressing a MRHA phenotype were identified and 1 such clone was used to subclone smaller DNA fragments able to confer the same MRHA and adherence properties as the parent strain. A 16.4 kilobase chromosomal DNA fragment cloned in pBR322 (pGTH1) expressed MRHA of human erythrocytes, adhered to HEp-2 cells and to human colonocyte brush borders. The identity of the cloned adhesin was confirmed by biochemical, genetic, electron microscopical and immunological comparison with the adhesin synthesized by 469-3.