Analysis of pilin polymerization in a uropathogenic Escherichia coli strain containing two pyelonephritis-associated (pap)gene clusters.

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Publication Type thesis
School or College School of Medicine
Department Pathology
Author Wilkins, Diana Gaudin.
Title Analysis of pilin polymerization in a uropathogenic Escherichia coli strain containing two pyelonephritis-associated (pap)gene clusters.
Date 1988-06
Description E. coli strain C1212 expresses two distinct pili on its cell surface that is encoded by separate pyelonephritis-associated pili (pap) gene clusters. These gene clusters have been demonstrated to produce expression of pili composed of either 17 kilodalton (kd) or 21 pilin monomers. The potential for copolymerization of the two pilin monomers, resulting in production of hybrid pili, was examined here. Pilin structural genes (A-B) and assembly genes (C-G) from each cluster were isolated and ligated to pBR325 and pACYC184 plasmid vectors, respectively. These vectors, harboring specific pap genes, were utilized to produce organisms carrying a set of pilin structural genes from one pap gene cluster and a set of assembly genes from the other (heterologous) pap gene cluster. Pilin monomers expressed by these organisms were examined by immunoelectron microscopy with 6nm and 18nm-diameter gold particles conjugated to a pap monoclonal antibody. Individual pilin monomers were also harvested and purified from overnight cultures of uropathogenic E. coli expressing either pilin-17 or pilin-21. Copolymerization of the two monomers was examined (in vitro) through specific iodination of pilin monomers, incubation and polymerization of monomer mixtures with 0.1M MgCl2, column chromatography of mixtures on Sepharose CL-6B and Sephadex G-100, and Immunoblots of pilin-17 and pilin-21 immunoprecipitates. Data gathered from complementation analysis of heterologous gene clusters and immunoprecipitates suggest that in vitro copolymerization of pilin monomers and hybrid pilin production can occur.
Type Text
Publisher University of Utah
Subject Microbiology; Urinary Organs; Polymers; Urinary Tract Infections
Subject MESH Pyelonephritis; Escherichia coli; Fimbriae, Bacterial; Genes, Bacterial
Dissertation Institution University of Utah
Dissertation Name MS
Language eng
Relation is Version of Digital reproduction of "Analysis of pilin polymerization in a uropathogenic Escherichia coli strain containing two pyelonephritis-associated (pap)gene clusters." Spencer S. Eccles Health Sciences Library. Print version of "Analysis of pilin polymerization in a uropathogenic Escherichia coli strain containing two pyelonephritis-associated (pap)gene clusters." available at J. Willard Marriott Library Special Collection. QR6.5 1988 .W54.
Rights Management © Diana Gaudin Wilkins.
Format Medium application/pdf
Identifier us-etd2,203
Source Original: University of Utah Spencer S. Eccles Health Sciences Library (no longer available).
ARK ark:/87278/s6q81tht
Setname ir_etd
Date Created 2012-04-23
Date Modified 2021-05-06
ID 192149
Reference URL https://collections.lib.utah.edu/ark:/87278/s6q81tht
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