Molecular cloning and characterisation of the genes for a non-fimbrial adhesin from Escherichia coli.
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 degrees C, but not 18 degrees 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 gamma delta mutagenesis. The NFA-1 operon was localised to a 6.5kb region of this fragment.
Hales
B A
BA
Department of Microbiology, University of Leicester, U.K.
Beverley-Clarke
H
H
High
N J
NJ
Jann
K
K
Perry
R
R
Goldhar
J
J
Boulnois
G J
GJ
D018830:Adhesins, Escherichia coli; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D003360:Cosmids; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D006386:Hemagglutination Tests; D006801:Humans; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D014170:Transformation, Genetic
D018830:Adhesins, Escherichia coli; D001425:Bacterial Outer Membrane Proteins