Infection and Immunity · 1983 · 179 citations · 31 references
BiosynthesisEngineeringStructural GeneVirulence FactorProtein BiosynthesisMolecular BiologyMolecular GeneticsDigalactoside-binding AdhesinMicrobiologyMolecular MicrobiologyMedicinePap PiliPap Pili MonomerBiomolecular EngineeringMicrobial Genetics
The uropathogenic strain Escherichia coli J96 mediates mannose-resistant hemagglutination owing to production of a digalactoside-binding adhesin. A cosmid clone from this strain has been isolated that, when harbored in E. coli K-12, expressed Pap pili and this adhesin (R. Hull et al., Infect. Immun. 33:933-938, 1981). By transposon mutagenesis and by the construction of a number of hybrid plasmid derivatives, we have demonstrated that about 8.5 kilobases of DNA is required to generate a mannose-resistant hemagglutination-positive phenotype in E. coli K-12 strain P678-54. The structural gene for the Pap pili monomer, papA, has been identified and mapped close to the promotor-proximal end of the Pap operon. Although strain P678-54 that harbored a Tn5 insertion within papA showed a mannose-resistant hemagglutination-positive phenotype, it was negative in a competitive enzyme-linked immunosorbent assay with anti-Pap pilus serum. This could mean that a Pap adhesin is encoded by a region on the Pap operon that is distinct from papA.
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A rapid alkaline extraction procedure for screening recombinant plasmid DNA
H.C. Birnboim, Janine Doly · Nucleic Acids Research · 1979 · 14.8K citations · Full text
Hakon Leffler, C. Svanborg Edén · FEMS Microbiology Letters · 1980 · 514 citations · Full text
Chemical Identification, Biochemistry, Microbial Pathogens +9