Difference between revisions of "Part:BBa K2336037"
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2336037 short</partinfo>
 
<partinfo>BBa_K2336037 short</partinfo>
Line 6: Line 5:
  
 
<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. When we use LBT exchange the Fe3+ binding site in the PmrB. Then the whole sensing system from Salmonella can sense the concentration of lanthanide ions in surroundings. 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.
+
LBT（lanthanide binding tag） is a kind of protein which can bind with the lanthanide ions. When we use LBT exchange the Fe3+ binding site in the PmrB. 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.
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
Line 13: Line 12:
  
 
<h1>'''Characterization'''</h1>
 
<h1>'''Characterization'''</h1>
−
This is a part for sensing part. LBT can bind the lanthanide ions and cause the fluorescence by the sensing system which can show the concentration of lanthanide. In our experiment,we let the engineering bacterium expresse GFP which means that the lanthanide ions in the water be sensed.
+
This is a part for sensing part. LBT can bind the lanthanide ions and excite the 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.
  
 
<h1>'''Improvement'''</h1>
 
<h1>'''Improvement'''</h1>
−
As we didn’t have enough time, we couldn’t let all the 12 LBT be exchanged successfully. After competition of this year, HUST-China will get others LBTs to be exchanged and form the whole system. If we make it, we will link gthe sensing part and capturing part to form the whole circuit which can sense the concentration of lanthanide ions and capture the lanthanide ions in the meantime. When our engineering bacterium finish their task, they can be recycle by silicon board.
+
As we didn’t have enough time, we couldn’t let all the 12 LBT 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.

Revision as of 17:00, 31 October 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. When we use LBT exchange the Fe3+ binding site in the PmrB. 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. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2116
    Illegal XhoI site found at 2155
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1704
    Illegal AgeI site found at 2881
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2804
    Illegal SapI.rc site found at 691


Characterization

This is a part for sensing part. LBT can bind the lanthanide ions and excite the 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

As we didn’t have enough time, we couldn’t let all the 12 LBT 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.