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Abstract
| The bacterial species Escherichia coli is responsible for the majority of urinary
tract infections in humans, in part due to attachment of bacteria to uroepithelial
cells via type 1 fimbriae. This study focused on one gene involved in type 1
pilus expression, fimE. FimE regulates expression of type 1 fimbriae by changing
the position of the structural gene’s promoter to prevent transcription. To
determine what genes may be involved in regulating fimE, an E. coli strain
lacking fim and lac genes was transformed with a plasmid containing the fimE
promoter hooked to a promoterless lacZ gene. The aims of this study were:
1) to randomly insert transposons into genes within the genome of this strain,
2) to screen the mutants generated for their ability to affect fimE expression as
measured by the amount of b-galactosidase produced, and 3) to identify by
DNA sequencing one or more genes affecting fimE expression. A total of 554
random transposon mutants were generated and screened for their ability to
affect fimE expression. Of these mutants, three reproducibly showed downregulation
of fimE, and one showed up-regulation. Further testing will be needed
to characterize the nature of their behavior, and eventual DNA sequencing
will elucidate which exact genes were mutated. |
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