Difference between revisions of "Part:BBa K2467000"
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To improve part BBa_K1659211, DsbADspB (dispersin exported to the extracellular using the DsbA signal sequence (KKIWLALAGLVLAFSASA)), we fused it to emGFP in order to measure secreted protein using fluorescence. We amplified the DsbADspB fragment and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emGFP vector which resulted in fluorescently tagged DspADspB. This fragment will then be moved to pSB1C3. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB. | To improve part BBa_K1659211, DsbADspB (dispersin exported to the extracellular using the DsbA signal sequence (KKIWLALAGLVLAFSASA)), we fused it to emGFP in order to measure secreted protein using fluorescence. We amplified the DsbADspB fragment and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emGFP vector which resulted in fluorescently tagged DspADspB. This fragment will then be moved to pSB1C3. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB. | ||
| − | Projects dealing with disruption of Biofilms can easily follow the release of the dispersin enzyme into the extracellular environment to degrade the biofilm polysaccharides | + | Projects dealing with disruption of Biofilms can easily follow the release of the dispersin enzyme into the extracellular environment to degrade the biofilm polysaccharides |
| + | |||
| + | The degradation function of DspB-MagR, immobilized DspB-MagR and Fe3O4@SiO2 was tested on the biofilm of staphylococcus aureus. Fe3O4@SiO2 showed no effect to detaching bacterial biofilms, DspB-MagR was able to remove 10% while the loaded DspB-MagR was able to remove 50%. The immobilized protein was also tested for biofilms of other bacterium like the Bacillus Cereus and Staphylococcus sp and showed significant removal rate. These results proved that using Fe3O4@SiO2 to immobilize the DspB-MagR recombinant protein is an alternative method for biofilm removal. | ||
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Revision as of 13:22, 20 October 2021
DsbA DspB emGFP
To improve part BBa_K1659211, DsbADspB (dispersin exported to the extracellular using the DsbA signal sequence (KKIWLALAGLVLAFSASA)), we fused it to emGFP in order to measure secreted protein using fluorescence. We amplified the DsbADspB fragment and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emGFP vector which resulted in fluorescently tagged DspADspB. This fragment will then be moved to pSB1C3. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB. Projects dealing with disruption of Biofilms can easily follow the release of the dispersin enzyme into the extracellular environment to degrade the biofilm polysaccharides
The degradation function of DspB-MagR, immobilized DspB-MagR and Fe3O4@SiO2 was tested on the biofilm of staphylococcus aureus. Fe3O4@SiO2 showed no effect to detaching bacterial biofilms, DspB-MagR was able to remove 10% while the loaded DspB-MagR was able to remove 50%. The immobilized protein was also tested for biofilms of other bacterium like the Bacillus Cereus and Staphylococcus sp and showed significant removal rate. These results proved that using Fe3O4@SiO2 to immobilize the DspB-MagR recombinant protein is an alternative method for biofilm removal.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 316
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 472