PMID:20351142

Citation	Puttamreddy, S, Cornick, NA and Minion, FC (2010) Genome-wide transposon mutagenesis reveals a role for pO157 genes in biofilm development in Escherichia coli O157:H7 EDL933.Infect. Immun. 78:2377-84
Abstract	Enterohemorrhagic Escherichia coli O157:H7, a world-wide human food-borne pathogen, causes mild to severe diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. The ability of this pathogen to persist in the environment contributes to its dissemination to a wide range of foods and food processing surfaces. Biofilms are thought to be involved in persistence, but the process of biofilm formation is complex and poorly understood in E. coli O157:H7. To better understand the genetics of this process, a mini-Tn5 transposon insertion library was constructed in strain EDL933 and screened for biofilm-negative mutants using a microtiter plate assay. Ninety-five of 11,000 independent insertions (0.86%) were biofilm negative, and transposon insertions were located in 51 distinct genes/intergenic regions that must be involved either directly or indirectly in biofilm formation. All of the 51 biofilm-negative mutants showed reduced biofilm formation on both hydrophilic and hydrophobic surfaces. Thirty-six genes were unique to this study, including genes on the virulence plasmid pO157. The type V secreted autotransporter serine protease EspP and the enterohemolysin translocator EhxD were found to be directly involved in biofilm formation. In addition, EhxD and EspP were also important for adherence to T84 intestinal epithelial cells, suggesting a role for these genes in tissue interactions in vivo.
Links	PubMed Online version:10.1128/IAI.00156-10
Keywords	Biofilms; Cell Adhesion; Cell Line; DNA Transposable Elements; Epithelial Cells; Escherichia coli O157; Escherichia coli Proteins; Genes, Bacterial; Genetic Complementation Test; Humans; Mutagenesis, Insertional; Plasmids
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Main Points of the Paper

Please summarize the main points of the paper.
The goal of this study was to gain additional insights into biofilm formation in E. coli O157:H7. A global mutational approach with a mini-Tn5 transposon was used to study the process of biofilm formation in strain EDL933, a strong bio- film-forming strain.

Materials and Methods Used

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Phenotype Annotations

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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: O157:H7 Substrain: EDL933	Genotype of Reference Strain:espP Genotype of Experimental Strain : Genotype of Reference Strain:espP	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: O157:H7 Substrain: EDL933	Genotype of Reference Strain:hisD	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: O157:H7 Substrain: EDL933	Genotype of Reference Strain: acrE	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Taxon: Escherichia coli O157:H7 EDL933	Genotype of Reference Strain: yaiH	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Taxon: Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:ynaJ	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:ecpD	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:aroC	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:cls	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:hlyB	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:glpQ	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:relA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:waaL	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:wbdP/per	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:waaD	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:ydeH	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:csgB/csgA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:yfeA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:waaJ	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:csgG	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:galU/hns	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:manC/wbdQ	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:csgA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:tolQ	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:waaP	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:hlyC/hlyA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:ychM	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:yahE/yahF	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:fcl	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:ybgC	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
a mutation or genetic difference within a strain	Escherichia coli O157:H7 EDL933	Genotype of Reference Strain:dsbA	Reference Condition:		biofilm formation			its transposon mutant shows biofilm-negative phenotype
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References

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