Difference between revisions of "Part:BBa K2570011"
 
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<partinfo>BBa_K2570011 short</partinfo>
 
<partinfo>BBa_K2570011 short</partinfo>
  
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In our project, we combined temperature promoter dsrA and salt-concentration promoter ProU as a logical gate controlled by salt-concentration and temperature to realize the aim of producing antibacterial, PLA.
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In our project, we combined temperature promoter ''dsrA''[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2570015 BBa_K2570015] and salt-concentration promoter ''proU''[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2467001 BBa_K2467001] as a logical gate controlled by salt-concentration and temperature to realize the aim of producing antibacterial, PLA.
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dsrA is a temperature control promoter. The temperature range of dsrA is from 25 degree to 42 degree. With temperature increase, the promote efficiency is getting higher. [1]The salt-concentration promoter ProU provides sensitivity to the salt concentration to human skin. ProU is a promoter existed in E.coli, induced by low salt concentration. And the HNS is a global-regulated protein which could bind to genome and specifically inhibit ProU.[2]
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''dsrA''[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2570015 BBa_K2570015] is a temperature control promoter. The temperature range of ''dsrA'' is from 25 degree to 42 degree. With temperature increase, the promote efficiency is getting higher. [1]The salt-concentration promoter ''proU''[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2467001 BBa_K2467001] provides sensitivity to the salt concentration to human skin. ''proU'' is a promoter existed in ''E.coli'', induced by low salt concentration. And the H-NS[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2570016 BBa_K2570016] is a global-regulated protein which could bind to genome and specifically inhibit ''proU''.[2]
  
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In our design, our engineering bacteria usually put in a spray container and store at room temperature. When temperature stays below 37 degree, the temperature promoter will not work.  
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In our design, our engineering bacteria usually put in a lotion container and store at room temperature. When temperature stays below 37 degree, the temperature promoter will not work.  
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[[File:T--FJNU-China--room_temperature.gif|600px|thumb|center|Fig.1 Compared with the negative control (the empty pSB1C3 vector).]]
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[[File:T--FJNU-China--room_temperature.jpg|600px|thumb|center|Fig.1 our circuits in room temperature.]]
  
  
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The function of PlacI has became a constitutive promoter. The global regulate protein HNS will be express to inhibit proU. Then the logical gate will be “off” state.
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The function of ''PlacI'' [https://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 BBa_R0011] has become a constitutive promoter. The global regulate protein H-NS will be expressed to inhibit ''proU''. Then the logical gate will be “off” state.
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On the other hand, when we use the spray on the skin, temperature reached 37 degree, the repressor protein lacI will be expressed to inhibit PlacI, the HNS won’t be revealed. Then, once people sweat, the Na+ and Cl- will induce proU, the logical gate became “on” state.
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[[File:T--FJNU-China--on_skin.gif|600px|thumb|center|Fig.1 Compared with the negative control (the empty pSB1C3 vector).]]
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On the other hand, when we use the lotion on the skin, temperature reached 37 degree, the repressor protein LacI [https://parts.igem.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012] will be expressed to inhibit ''PlacI'', the H-NS won’t be revealed. Then, once people sweat, the Na+ and Cl- will induce ''proU'', the logical gate became “on” state.
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[[File:T--FJNU-China--on_skin.jpg|600px|thumb|center|Fig.1 our circuits on people skin.]]
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Firstly we use GFP to characterize the function of our circuit. And after finishing the construction of the whole circuits, we set 3 gradient of salt concentration, 0M, 0.05M, 0.3M, and 3 gradient of temperature, 25℃, 32.5℃ and 37℃, detected the fluorescence level and draw the line of different culture conditions.
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[[File:T--FJNU-China--flourencence_level_different_salt_and_tem_2.png|600px|thumb|center|Fig.1 fluorescence level at different salt concentration and temperature.]]
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[[File:T--FJNU_China--flourensence_line_bar.jpg|600px|thumb|center|Fig.2 the line of fluorescence level.]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 09:04, 15 October 2018


dsrA+lacI+PlacI+hns+ProU+GFP

In our project, we combined temperature promoter dsrABBa_K2570015 and salt-concentration promoter proUBBa_K2467001 as a logical gate controlled by salt-concentration and temperature to realize the aim of producing antibacterial, PLA. dsrABBa_K2570015 is a temperature control promoter. The temperature range of dsrA is from 25 degree to 42 degree. With temperature increase, the promote efficiency is getting higher. [1]The salt-concentration promoter proUBBa_K2467001 provides sensitivity to the salt concentration to human skin. proU is a promoter existed in E.coli, induced by low salt concentration. And the H-NSBBa_K2570016 is a global-regulated protein which could bind to genome and specifically inhibit proU.[2]

In our design, our engineering bacteria usually put in a lotion container and store at room temperature. When temperature stays below 37 degree, the temperature promoter will not work.

Fig.1 our circuits in room temperature.


The function of PlacI BBa_R0011 has become a constitutive promoter. The global regulate protein H-NS will be expressed to inhibit proU. Then the logical gate will be “off” state.

On the other hand, when we use the lotion on the skin, temperature reached 37 degree, the repressor protein LacI BBa_C0012 will be expressed to inhibit PlacI, the H-NS won’t be revealed. Then, once people sweat, the Na+ and Cl- will induce proU, the logical gate became “on” state.

Fig.1 our circuits on people skin.

Firstly we use GFP to characterize the function of our circuit. And after finishing the construction of the whole circuits, we set 3 gradient of salt concentration, 0M, 0.05M, 0.3M, and 3 gradient of temperature, 25℃, 32.5℃ and 37℃, detected the fluorescence level and draw the line of different culture conditions.

Fig.1 fluorescence level at different salt concentration and temperature.
Fig.2 the line of fluorescence level.


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 1587
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3324
    Illegal SapI.rc site found at 154
    Illegal SapI.rc site found at 1973