Difference between revisions of "Part:BBa K2377009"

 
 
(5 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2377009 short</partinfo>
 
<partinfo>BBa_K2377009 short</partinfo>
  
one element to sense zinc ion
+
Improved target:
 +
BBa_K1755301
 +
 
 +
Improved form:
 +
 
 +
We replaced a sensor element(Copper-Responsive Two-Component System BBa_K190017)for the Zinc-Responsive Two-Component regulatory system (contains σ54 promoter).
 +
 
 +
Mechanism of changed sensor
 +
The mechanism of Zinc sensor is related to a intrinsic signaling pathway in Ecoli.
 +
Which is known that many sensing proteins ZraS immobilized on the membrane of cell.
 +
And when Zn2+ show up in periplasmic space, the ions will combine with ZraS and trigger it autophosphorylation. (ZraS*)
 +
The phosphorylation signal will transmit from ZraS* to ZraR, then ZraR* come to ZraR(HydG) binding site[1] that will make σ54promoter start downstream transcription.
 +
(ZraS* ,ZraR*  means activated condition.)
 +
 +
https://static.igem.org/mediawiki/parts/thumb/2/21/Lanzhou_Parts_2.png/800px-Lanzhou_Parts_2.png
 +
 
 +
So, we substitute Zinc-Responsive Two-Component regulatory system for the copper one in BBa_K1755301. Successfully construct a new heavy metal sensor.
 +
 
 +
Function of improved new part
 +
Improved new part is BBa_K2377009, which can detect the concentration of Zn2+ and transform it into electric signal.
 +
 
 +
The downstream protein ribB (3,4-dihydroxy-2-butanone 4-phosphate synthase ,ribB) could catalyze the conversion of D-ribulose 5-phosphate formate to 3,4-dihydroxy-2-butanone 4-phosphate. This step is part of the biosynthesis pathway of riboflavin, an electron carrier in microfunction circuit (MFC).
 +
 
 +
So when the whole gene circuit works, it will result in a consequence that once the concentration of Zn2+ in environment increased, more riboflavin will be produced to impact the voltage of MFC.
 +
 
 +
Through this new part we realized monitoring heavy metal Zn2+ automatically, quantitatively and efficiently.
 +
 
 +
The figure below shows the comparison of the part BBa_K1755301 (left) with the new improved part BBa_K2377009 (right), the riboflavin concentration versus heavy metal ion concentration.
 +
 
 +
https://static.igem.org/mediawiki/parts/thumb/e/e9/Lanzhou_Parts_1.png/800px-Lanzhou_Parts_1.png
 +
 
 +
And this is the improve target biobrick designed by 2015 LZU-China iGEM team
 +
 
 +
https://parts.igem.org/Part:BBa_K1755301
 +
 
 +
https://parts.igem.org/Part:BBa_K1755303
 +
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
 +
  
 
<!-- -->
 
<!-- -->

Latest revision as of 22:59, 31 October 2017


zrap ribB

Improved target: BBa_K1755301

Improved form:

We replaced a sensor element(Copper-Responsive Two-Component System BBa_K190017)for the Zinc-Responsive Two-Component regulatory system (contains σ54 promoter).

Mechanism of changed sensor The mechanism of Zinc sensor is related to a intrinsic signaling pathway in Ecoli. Which is known that many sensing proteins ZraS immobilized on the membrane of cell. And when Zn2+ show up in periplasmic space, the ions will combine with ZraS and trigger it autophosphorylation. (ZraS*) The phosphorylation signal will transmit from ZraS* to ZraR, then ZraR* come to ZraR(HydG) binding site[1] that will make σ54promoter start downstream transcription. (ZraS* ,ZraR* means activated condition.)

800px-Lanzhou_Parts_2.png

So, we substitute Zinc-Responsive Two-Component regulatory system for the copper one in BBa_K1755301. Successfully construct a new heavy metal sensor.

Function of improved new part Improved new part is BBa_K2377009, which can detect the concentration of Zn2+ and transform it into electric signal.

The downstream protein ribB (3,4-dihydroxy-2-butanone 4-phosphate synthase ,ribB) could catalyze the conversion of D-ribulose 5-phosphate formate to 3,4-dihydroxy-2-butanone 4-phosphate. This step is part of the biosynthesis pathway of riboflavin, an electron carrier in microfunction circuit (MFC).

So when the whole gene circuit works, it will result in a consequence that once the concentration of Zn2+ in environment increased, more riboflavin will be produced to impact the voltage of MFC.

Through this new part we realized monitoring heavy metal Zn2+ automatically, quantitatively and efficiently.

The figure below shows the comparison of the part BBa_K1755301 (left) with the new improved part BBa_K2377009 (right), the riboflavin concentration versus heavy metal ion concentration.

800px-Lanzhou_Parts_1.png

And this is the improve target biobrick designed by 2015 LZU-China iGEM team

https://parts.igem.org/Part:BBa_K1755301

https://parts.igem.org/Part:BBa_K1755303


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 625
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 515
    Illegal AgeI site found at 557
  • 1000
    COMPATIBLE WITH RFC[1000]