Difference between revisions of "Part:BBa K5127014"
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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). | 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 | + | |
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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). | 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). | ||
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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"/> | <p style="text-align:center;"><img src="https://static.igem.wiki/teams/5127/parts/phdph-ci-1.jpg" width="400" height="auto"/> | ||
<br> | <br> | ||
| − | Figure 1. Plasmid design of HpdR-pHpdH-cI-GFP Created by biorender.com. | + | <i>Figure 1. Plasmid design of HpdR-pHpdH-cI-GFP Created by biorender.com.</i> |
</p> | </p> | ||
| + | </html> | ||
<HTML> | <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"/> | <p style="text-align:center;"><img src="https://static.igem.wiki/teams/5127/parts/phdph-ci-2.jpg" width="400" height="auto"/> | ||
<br> | <br> | ||
| − | Figure 2. The agarose gel electrophoresis results of the PCR products of pHpdH-CI construction. The materials used to construct pHpdH-CI. | + | <i>Figure 2. The agarose gel electrophoresis results of the PCR products of pHpdH-CI construction. The materials used to construct pHpdH-CI.</i> |
</p> | </p> | ||
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| − | + | ===Sequence and Features=== | |
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<partinfo>BBa_K5127014 SequenceAndFeatures</partinfo> | <partinfo>BBa_K5127014 SequenceAndFeatures</partinfo> | ||
Latest revision as of 12:41, 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.
Currently, we are still constructing this part and have not yet obtained the kinetics measurement.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1237
Illegal NotI site found at 408 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 392
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2386
- 1000COMPATIBLE WITH RFC[1000]