Difference between revisions of "Part:BBa K2336039"
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<h1>'''Usage and biology'''</h1> | <h1>'''Usage and biology'''</h1> | ||
| − | LBT（lanthanide binding tag） is a kind of protein which can bind with the lanthanide ions. We exchange the Fe3+ binding site in pmrB system with LBT. Then the whole sensing system from Salmonella can sense the concentration of lanthanide ions nearby. When the concentration of lanthanide ions reach the threshold, the lanthanide can be bound with LBT. Then the pmrB will activate pmrA by phosphorylating it. Phosphorylated PmrA is going to activate promoter pmrC in next step. In the end, GFP behind pmrC will be expressed. You can get the relative concentration of lanthanide according to the fluorescence intensity. | + | <p>LBT（lanthanide binding tag） is a kind of protein which can bind with the lanthanide ions. We exchange the Fe3+ binding site in pmrB system with LBT. Then the whole sensing system from Salmonella can sense the concentration of lanthanide ions nearby. When the concentration of lanthanide ions reach the threshold, the lanthanide can be bound with LBT. Then the pmrB will activate pmrA by phosphorylating it. Phosphorylated PmrA is going to activate promoter pmrC in next step. In the end, GFP behind pmrC will be expressed. You can get the relative concentration of lanthanide according to the fluorescence intensity.Figure 1 shows the circuit of PmrA-PmrB(LBT11)-PmrC-GFP. |
| − | + | [[File:ABC.png|400px|thumb|center|Figure1:PmrA-PmrB(LBT11)-PmrC-GFP]] | |
| − | + | </p> | |
| − | + | ||
| − | + | ||
<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
| − | This is a part for sensing part. LBT11 can bind the lanthanide ions and excite fluorescence by the sensing system, which can show the concentration of lanthanide. In our experiment, we let the engineering bacterium express GFP, which means that the lanthanide ions in the water can be sensed. | + | <p>This is a part for sensing part. LBT11 can bind the lanthanide ions and excite fluorescence by the sensing system, which can show the concentration of lanthanide. In our experiment, we let the engineering bacterium express GFP, which means that the lanthanide ions in the water can be sensed.</p> |
<h1>'''Improvement'''</h1> | <h1>'''Improvement'''</h1> | ||
| − | We designed 12 LBTs to exchange the iron ion binding site, because we aim to test their binding force to select the most sensitive system. As we didn’t have enough time, we couldn’t let all the 12 LBTs be exchanged successfully. After competition of this year, HUST-China will get other LBTs exchanged to form the whole system. If we make it, we will link the sensing part and capture part to form the whole circuit which can sense the concentration of lanthanide ions and capture the lanthanide ions right after it. When our engineering bacteria finish their task, they can be recycle by silicon board. | + | <p>We designed 12 LBTs to exchange the iron ion binding site, because we aim to test their binding force to select the most sensitive system. As we didn’t have enough time, we couldn’t let all the 12 LBTs be exchanged successfully. After competition of this year, HUST-China will get other LBTs exchanged to form the whole system. If we make it, we will link the sensing part and capture part to form the whole circuit which can sense the concentration of lanthanide ions and capture the lanthanide ions right after it. When our engineering bacteria finish their task, they can be recycle by silicon board.</p> |
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| + | <!-- --> | ||
| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K2336037 SequenceAndFeatures</partinfo> | ||
Revision as of 12:51, 1 November 2017
PmrA-PmrB(LBT5)-PmrC-GFP
The sensing part of our circuit. LBT will be a part of membrane protein pmrB. The lanthanon can be bound with LBT. Then pmrA will be phosphorylated and activate pmrC. In the end, GFP behind pmrC will be expressed.
Usage and biology
LBT（lanthanide binding tag） is a kind of protein which can bind with the lanthanide ions. We exchange the Fe3+ binding site in pmrB system with LBT. Then the whole sensing system from Salmonella can sense the concentration of lanthanide ions nearby. When the concentration of lanthanide ions reach the threshold, the lanthanide can be bound with LBT. Then the pmrB will activate pmrA by phosphorylating it. Phosphorylated PmrA is going to activate promoter pmrC in next step. In the end, GFP behind pmrC will be expressed. You can get the relative concentration of lanthanide according to the fluorescence intensity.Figure 1 shows the circuit of PmrA-PmrB(LBT11)-PmrC-GFP.
Characterization
This is a part for sensing part. LBT11 can bind the lanthanide ions and excite fluorescence by the sensing system, which can show the concentration of lanthanide. In our experiment, we let the engineering bacterium express GFP, which means that the lanthanide ions in the water can be sensed.
Improvement
We designed 12 LBTs to exchange the iron ion binding site, because we aim to test their binding force to select the most sensitive system. As we didn’t have enough time, we couldn’t let all the 12 LBTs be exchanged successfully. After competition of this year, HUST-China will get other LBTs exchanged to form the whole system. If we make it, we will link the sensing part and capture part to form the whole circuit which can sense the concentration of lanthanide ions and capture the lanthanide ions right after it. When our engineering bacteria finish their task, they can be recycle by silicon board.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2116
Illegal XhoI site found at 2155 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1704
Illegal AgeI site found at 2881 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2804
Illegal SapI.rc site found at 691