Team:Tokyo Tech/Project
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- | [[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assay.png| | + | [[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assay.png|200px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] |
- | [[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assayparts.png| | + | [[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assayparts.png|300px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] |
[[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assaydeta.png|200px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] | [[File:gene circuit positive feedback tokyotech 3OC12HSL→pLasLuXI-PLux assaydeta.png|200px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] | ||
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==positive feedback assay== | ==positive feedback assay== | ||
[[File:gene circuit positive feedback tokyotech positive feedback assay.png|300px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] | [[File:gene circuit positive feedback tokyotech positive feedback assay.png|300px|thumb|left|fig5,3OC6HSL→pLuxLasI-PLas]] |
Revision as of 09:11, 3 September 2012
Project OVERVIEW
story
The stage is city Verona, Italy in the 14th century.Romeo met Juliette. Two persons fall to love instantly.Romeo will be involved in argument of a street with a friend, and Romeo which flew into a rage over the best friend having been killed will kill Capulet wife's nephew. The grand duke of Verona carries out Romeo on the crime of banishment. Lawrence who was able to ask Juliette for help stands the trick using the poison of suspended animation so that he may mate her with Romeo. However, this plan did not get across to banished Romeo well. Therefore, Juliette who Romeo which thought that Juliette died drank poison at her grave, and woke up to death from a state of suspended animation just behind that also follows the back with the dirk of Romeo.
Gene circuit
In this project,we define the signal which E.coli takes out as the romantic feeling of Romeo and Juliette.
The story which we reproduce is divided into three stages.
First,in positive feedback circuit, there are two types of E. coli, Cell R(Romeo) and Cell J(Juliette). In cell R (fig2), production of LasI is activated by LuxR/3OC6HSL complex and In cell J (fig2), production of LuxI is activated by LasR/3OC12HSL complex. This means that 3OC12HSL produced by cell R and 3OC6HSL produced by cell J indirectly activate each other’s production. we express the story of "Romeo met Juliette. Two persons fall to love instantly.", using "positive feedback circuit" as love which burns vigorously.
Second,"band detect system(fig3)" is a mechanism which controls production of a signal in Cell J body, when the amount of secretion of the signal by Cell R reaches more than fixed. we express the story of "Juliette for help stands the trick using the poison of suspended animation so that he may mate her with Romeo.", using a "band detect system" as Juliette's suspended animation .
Third,if Cell R detects reduction of the signal from Cell J, control of a suicide gene will separate in Cell R body. As a result, the signals which Cell R produces in connection with reduction and it in the number of individuals of Cell R decrease in number. Finally, by the same system as Cell R, a suicide gene also reveals Cell J and signals further decrease in number. we express the story of "Juliette who Romeo which thought that Juliette died drank poison at her grave, and woke up to death from a state of suspended animation just behind that also follows the back with the dirk of Romeo.", using a suicide mechanism as suicide of Romeo and Juliette.
Finaly,we argued about feasibility by performing a simulation using modeling about whether all the above stories are materialized.
Result
In order to make the story of "Romeo and Juliette", we constructed and characterized several important parts and devices. First we characterized (Parts name), both of which are existing parts. Second, we designed and characterized a new (parts name). Third, we designed and characterized a NEW BioBrick device (Parts name),(explain). Forth, we confirmed that (mechanism). Finally, we simulated our project and confirmed the feasibility of our system.
Positive feedback mechanism
LasR generator
3OC6HSL→pLuxLasI-PLas
3OC12HSL→pLasLuXI-PLux
positive feedback assay
Band detect system
Lux-Tet hybrid promoter assay
Suicide mechanism
Las→Lac assay, Lux→Lac assay
modeling
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