Cloning of genes determining the production of mannose-resistant fimbriae in a uropathogenic strain of Escherichia coli belonging to serogroup O6.
Chromosomal DNA from a uropathogenic strain of Escherichia coli was partially digested with the restriction enzyme EcoRI. The partial digests were ligated into a cosmid containing an ampicillin-resistant determinant and packaged into lambda phage particles. An ampicillin-resistant transductant of E. coli HB101 was found to possess mannose-resistant hemagglutinating activity associated with a 50-kilobase-pair plasmid. Subcloning of the mannose-resistant fimbrial genes revealed that the genetic determinants were encoded by a 6.9-kilobase-pair DNA fragment of a recombinant plasmid. Chimeric plasmids smaller in size were unable to transform E. coli to fimbrial production. Physical maps of the recombinant plasmids were prepared showing restriction endonuclease sites within the inserted DNA fragments. The hemagglutinating activities of the wild-type strain and of the recombinant derivative were compared. Both strains agglutinated human erythrocytes in the presence of D-mannose to the same degree and also failed to produce fimbriae after incubation at 18 degrees C. Also, both strains were agglutinated by antifimbrial serum at a high titer, whereas no such activity was observed when a strain of E. coli which did not possess a plasmid was used.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics* / Q000648:ultrastructure; D010861:Fimbriae, Bacterial; D005798:Genes, Bacterial; D006384:Hemagglutination; D010957:Plasmids
D004262:DNA Restriction Enzymes