A new locus, pilE, required for the binding of type 1 piliated Escherichia coli to erhythrocytes |
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Authors: | Lisa Maurer Paul E Orndorff |
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Institution: | Department of Microbiology, Pathology and Parasitology, North Carolina State University, School of Veterinary Medicine, 4700 Hillsborough Street, Raleigh, NC 27606, U.S.A. |
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Abstract: | Abstract We have identified a new locus required for mediating the binding of type 1 piliated Escherichia coli cells to guinea pig erythrocytes. This locus, pilE , was discovered after restrition site mutagenesis of a cloned segment of DNA containing the chromosomal pil region from a clinical strain of E. coli. pilE Mutants failed to agglutinate guinea pig erythrocytes but expressed pili were morphologically and antigenically indistinguishable from the parental ( pilE +) strain. Construction of a chromosomal pilE mutation in E. coli K-12 was accomplished by introducing a restriction fragment containing a pilE lesion into the chromosome of a recBC sbcB host strain. Mutations in pilE could be complimented in trans by the addition of a cloned segment of DNA containing the parental pilE locus. Lesions in any of the genes required for pilus assembly also produced a hemagglutination minus phenotype suggesting that both the product(s) specified by the pilE locus and pili were required for hemagglutination. Hemagglutination experiments using partially purified pili also supported this suggestion. |
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Keywords: | Pili adhesin agglutination site-directed mutagenesis recBC sbcB mutants |
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