Difference between revisions of "Part:BBa K5127014"
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This part is constructed for the characterization of the HpdR-cI-GFP butyrate biosensor in Nissle 1917. | This part is constructed for the characterization of the HpdR-cI-GFP butyrate biosensor in Nissle 1917. | ||
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| − | === | + | |
| + | ===Usage and Biology==== | ||
| + | When butyrate is present, it binds to HpdR, changing its conformation so it could bind and activate the pHpdH promoter, resulting in expression of downstream genes, in which GFP downstream of the plam promoter will be expressed if butyrate concentration is relatively low. This is an inverse logic of the part pHpdR (BBa_K2560304). | ||
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| + | ===Design and build of butyrate biosensor using pHpdH-cI==== | ||
| + | To integrate the butyrate biosensor into our ultimate project goal, we designed the CI gene into the pHpdH sensing system plasmid. In this system, when butyrate levels are low, the output of GFP will be higher (Figure 1). This design enables us to monitor and respond to butyrate concentration effectively within our system. We contructed this plasmid by using Golden Gate Assembly (Figure 1). | ||
| + | <HTML> | ||
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| + | <p style="text-align:center;"><img src="https://static.igem.wiki/teams/5127/parts/phdph-ci-1.jpg" width="400" height="auto"/> | ||
| + | <br> | ||
| + | Figure 1. Plasmid design of HpdR-pHpdH-cI-GFP Created by biorender.com. | ||
| + | </p> | ||
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| + | <HTML> | ||
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| + | <p style="text-align:center;"><img src="https://static.igem.wiki/teams/5127/parts/phdph-ci-2.jpg" width="400" height="auto"/> | ||
| + | <br> | ||
| + | Figure 2. The agarose gel electrophoresis results of the PCR products of pHpdH-CI construction. The materials used to construct pHpdH-CI. | ||
| + | </p> | ||
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| + | Currently, we are still constructing this part and have not yet obtained the kinetics measurement. | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K5127014 SequenceAndFeatures</partinfo> | <partinfo>BBa_K5127014 SequenceAndFeatures</partinfo> | ||
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Revision as of 08:07, 2 October 2024
Butyrate sensor with NOT gate (pHpdH-ci-HpdR)
This part is constructed for the characterization of the HpdR-cI-GFP butyrate biosensor in Nissle 1917.
Usage and Biology=
When butyrate is present, it binds to HpdR, changing its conformation so it could bind and activate the pHpdH promoter, resulting in expression of downstream genes, in which GFP downstream of the plam promoter will be expressed if butyrate concentration is relatively low. This is an inverse logic of the part pHpdR (BBa_K2560304).
Design and build of butyrate biosensor using pHpdH-cI=
To integrate the butyrate biosensor into our ultimate project goal, we designed the CI gene into the pHpdH sensing system plasmid. In this system, when butyrate levels are low, the output of GFP will be higher (Figure 1). This design enables us to monitor and respond to butyrate concentration effectively within our system. We contructed this plasmid by using Golden Gate Assembly (Figure 1).
Figure 1. Plasmid design of HpdR-pHpdH-cI-GFP Created by biorender.com.
Figure 2. The agarose gel electrophoresis results of the PCR products of pHpdH-CI construction. The materials used to construct pHpdH-CI.