Difference between revisions of "Part:BBa K4767013"
 
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Uses a factor Pars(BBa_K4767001) ,strong RBS(BBa_J34801), DNA binding transcriptional repressor <i>arsR</i>(BBa_J15101), <i>luxR</i>(△2-162)(BBa_K4767000) TT(BBa_B015) ,PluxⅠ(BBa_R0062) and <i>gfp</i>(BBa_E0040).This composite part is a BioBrick.
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Uses a factor P<i><sub>ars</sub></i>(BBa_K4767001) ,strong RBS(BBa_J34801), DNA binding transcriptional repressor <i>arsR</i>(BBa_J15101), <i>luxR</i>(△2-162)(BBa_K4767000) TT(BBa_B0015) ,P<sub><i>luxⅠ</i></sub>(BBa_R0062) and <i>gfp</i>(BBa_E0040).This composite part is a BioBrick.
  
  
 
===Usage and Biology===
 
===Usage and Biology===
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In order to construct a positive feedback circuit which does not require 3OC6HSL produced by LuxI, we engineered LuxR by deleting 2-262 amino acids in the N-terminal domain (AHL binding domain) and reserving a C-terminal domain with the function of activating transcription, obtaining a resulting regulator LuxR(△2-162).  LuxR(△2-162) can active the gene transcription driven by the <i>lux</i> promoter in the absence of AHL. To construct the amplifier, we cloned <i>gfp</i> and LuxR(Δ2-162) behind the <i>lux</i> promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the PluxI promoter and activate its own transcription.  
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In order to construct a positive feedback circuit which does not require 3OC6HSL produced by LuxI, we engineered LuxR by deleting 2-262 amino acids in the N-terminal domain (AHL binding domain) and reserving a C-terminal domain with the function of activating transcription, obtaining a resulting regulator LuxR(△2-162).  LuxR(△2-162) can active the gene transcription driven by the <i>lux</i> promoter in the absence of AHL. To construct the amplifier, we cloned <i>gfp</i> and LuxR(Δ2-162) behind the <i>lux</i> promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the P<sub><i>luxⅠ</i></sub> promoter and activate its own transcription.  
  
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<center>[[File:BBa_K4767013.jpg]]</center>
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<center>https://static.igem.wiki/teams/4767/wiki/part/img-1178.png</center>
  
 
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Latest revision as of 08:30, 8 October 2023


Pars-RBS-arsR-RBS-luxR(△2-162)-TT-PluxⅠ-RBS-luxR(△2-162)-RBS-gfp -TT


Uses a factor Pars(BBa_K4767001) ,strong RBS(BBa_J34801), DNA binding transcriptional repressor arsR(BBa_J15101), luxR(△2-162)(BBa_K4767000) TT(BBa_B0015) ,PluxⅠ(BBa_R0062) and gfp(BBa_E0040).This composite part is a BioBrick.


Usage and Biology

In order to construct a positive feedback circuit which does not require 3OC6HSL produced by LuxI, we engineered LuxR by deleting 2-262 amino acids in the N-terminal domain (AHL binding domain) and reserving a C-terminal domain with the function of activating transcription, obtaining a resulting regulator LuxR(△2-162). LuxR(△2-162) can active the gene transcription driven by the lux promoter in the absence of AHL. To construct the amplifier, we cloned gfp and LuxR(Δ2-162) behind the lux promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the PluxⅠ promoter and activate its own transcription.

img-1178.png

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 270
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 929
    Illegal BsaI.rc site found at 1952

References

Xiaoqiang Jia, Bu Rongrong, Zhao Tingting, et al. Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection[J]. Applied and environmental microbiology, 2019, 85(11): 1.

Nistala, G.J., et al., A modular positive feedback-based gene amplifier. J Biol Eng, 2010. 4: p. 4.