Difference between revisions of "Part:BBa K2377009"
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===Usage and Biology=== | ===Usage and Biology=== | ||
+ | Improved target: | ||
+ | BBa_K1755301(K1755024、K1755305、K1755303) | ||
+ | |||
+ | 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 | ||
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Revision as of 17:58, 27 October 2017
zrap ribB
one element to sense zinc ion
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 625
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 515
Illegal AgeI site found at 557 - 1000COMPATIBLE WITH RFC[1000]