Difference between revisions of "Part:BBa K3832007"

 
Line 3: Line 3:
 
<partinfo>BBa_K3832007 short</partinfo>
 
<partinfo>BBa_K3832007 short</partinfo>
  
−
This part is a kind of toggle-switch circuit, which can turn into bistable statement of protein expression in different inducing conditions. Two Promoter-Repressor gene systems (lacUV5 promoter-lacI and Plambda-CI857) are contained in this circuit. Once being induced by IPTG, the downstream genes of lacUV5 will expression stably, while those in the downstream of Plambda would be repressed. At temperatures above 42 degrees Celsius, gene expression moves into the opposite situation. With RFP CDS following the lacUV5 promoter and GFP CDS following the Plambda promoter, the effects of bistability can be more easily represented. Additional genes can be added after the GFP/RFP to achieve its bistable expression.
+
This part is an updated version of the traditional toggle-switch developed by James J. Collins's group in 2000(Gardner et al. Nature, 2000), which contains two feedbacks each controls the other, and can achieve bistability of protein expression in different inducing conditions.Two promoter-repressor systems (lacUV5 promoter-LacI and lambda-CI857) with sfGFP and mRFP respectively, was constructed in our updated version, to monitor the two states of the circuit. With induction of IPTG, the downstream genes of lacUV5, that is, CI protein and mRFP will expressed, while those in the downstream of lambda promoter (LacI and sfGFP) will be repressed. Even without IPTG induction after several hours, the lack of LacI expression will result in the stability of red fluorescence. At temperatures above 42 degrees Celsius, gene expression will be flipped into another state, the stable expression of LacI and sfGFP, and the state will maintain even without heat. The GFP and RFP can be altered with other functional genes such as tyrptohan synthesitic genes to achieve the bistable expression and synthesis of tyrptohan.
  
  

Revision as of 03:55, 16 October 2021


Toggle-switch circuit with GFP and RFP

This part is an updated version of the traditional toggle-switch developed by James J. Collins's group in 2000(Gardner et al. Nature, 2000), which contains two feedbacks each controls the other, and can achieve bistability of protein expression in different inducing conditions.Two promoter-repressor systems (lacUV5 promoter-LacI and lambda-CI857) with sfGFP and mRFP respectively, was constructed in our updated version, to monitor the two states of the circuit. With induction of IPTG, the downstream genes of lacUV5, that is, CI protein and mRFP will expressed, while those in the downstream of lambda promoter (LacI and sfGFP) will be repressed. Even without IPTG induction after several hours, the lack of LacI expression will result in the stability of red fluorescence. At temperatures above 42 degrees Celsius, gene expression will be flipped into another state, the stable expression of LacI and sfGFP, and the state will maintain even without heat. The GFP and RFP can be altered with other functional genes such as tyrptohan synthesitic genes to achieve the bistable expression and synthesis of tyrptohan.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2854
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2854
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2854
    Illegal AgeI site found at 1891
    Illegal AgeI site found at 3436
    Illegal AgeI site found at 3548
  • 1000
    COMPATIBLE WITH RFC[1000]