Difference between revisions of "Part:BBa K1850000"
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<partinfo>BBa_K1850000 short</partinfo> | <partinfo>BBa_K1850000 short</partinfo> | ||
| − | + | This part contains the ''fimH'' adhesin under control of a titratable rhamnose promoter. The FimH protein is a subunit of a naturally occurring structure in some strains of ''E. coli'' called Type 1 Pili. These hairlike appendages ypically manifest as organelles on the surface of pathogenic E. coli which are responsible for urinary tract infections in humans. The FimH adhesin is found at the tip of the pilus. This part can be cotransformed with <partinfo>BBa_K1850013</partinfo>, which contains the rest of the ''fim'' operon, and induced to produce type 1 pili. | |
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| + | We established that our engineered system could control pili expression by performing an agglutination (link to first description and graphics in strains and assays). Following our protocol, we mixed cultures of our induced and uninduced plasmid-containing fimB knockouts with S. cervisiae yeast along with controls. We found that our plasmids recovered agglutination in the negative control strain and that this agglutination was dependent on the addition of inducer molecules. This shows both that we have control over expression of the biosynthetic machinery and the fimH adhesin, and that the resulting pili were functional as determined by a standard assay for mannose binding. This assay was performed in biological triplicates with the same result. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 23:10, 18 September 2015
pRha - fimH
This part contains the fimH adhesin under control of a titratable rhamnose promoter. The FimH protein is a subunit of a naturally occurring structure in some strains of E. coli called Type 1 Pili. These hairlike appendages ypically manifest as organelles on the surface of pathogenic E. coli which are responsible for urinary tract infections in humans. The FimH adhesin is found at the tip of the pilus. This part can be cotransformed with BBa_K1850013, which contains the rest of the fim operon, and induced to produce type 1 pili.
We established that our engineered system could control pili expression by performing an agglutination (link to first description and graphics in strains and assays). Following our protocol, we mixed cultures of our induced and uninduced plasmid-containing fimB knockouts with S. cervisiae yeast along with controls. We found that our plasmids recovered agglutination in the negative control strain and that this agglutination was dependent on the addition of inducer molecules. This shows both that we have control over expression of the biosynthetic machinery and the fimH adhesin, and that the resulting pili were functional as determined by a standard assay for mannose binding. This assay was performed in biological triplicates with the same result.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]