Difference between revisions of "Part:BBa K5477043"

 
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<partinfo>BBa_K5477043 short</partinfo>
 
<partinfo>BBa_K5477043 short</partinfo>
  
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In this system, the pRET2-LexA-ER&#945;(LBD) receptor module responds to the presence of estrogen or estrogen-like compounds, in this case BPA. When these ligands bind to the ER&#945; LBD, it triggers a transcriptional response by allowing the LexA-ER&#945; chimeric activator to bind to the Lex6Op sequences in the pLex6Op-pLEU2-NanoLuc reporter module. This activates the pLEU2 promoter, leading to the expression of NanoLuc, which produces a luminescent signal. The intensity of the luminescence is proportional to the concentration of the estrogenic compound in the environment, providing a measurable output of receptor activation.
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===Summary===
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In this system, the pRET2-LexA-ERα(LBD) receptor module responds to the presence of estrogen or estrogen-like compounds, in this case BPA. When these ligands bind to the ERα LBD, it triggers a transcriptional response by allowing the LexA-ERα chimeric activator to bind to the Lex6Op sequences in the pLex6Op-pLEU2-NanoLuc reporter module. This activates the pLEU2 promoter, leading to the expression of NanoLuc, which produces a luminescent signal. The intensity of the luminescence is proportional to the concentration of the estrogenic compound in the environment, providing a measurable output of receptor activation.
  
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===Usage and Biology===
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In this biosensor system, the combination of receptor and reporter modules is designed to monitor the activity of Estrogen Receptor Alpha (ERα) in response to estrogenic compounds, such as estrogen or bisphenol A (BPA), and generate a luminescent signal through the expression of NanoLuc.
  
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<i>Why do we want to detect BPA in breast milk? </i>
  
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<h2>Receptor Module</h2>
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===Usage and Biology===
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1.pRET2-LexA-ERα(LBD) [https://parts.igem.org/Part:BBa_K5477027| BBa_K5477027]: This receptor module utilizes the pRET2 promoter to drive the expression of a fusion protein composed of the LexA DNA-binding domain (DBD) and the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ERα). The LexA-ERα(LBD) fusion allows the system to detect estrogen or estrogen-like molecules such as BPA. When BPA binds to the ERα LBD, it induces a conformational change in the receptor, activating the LexA DBD to bind to specific LexA operator sequences (Lex6Op).
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<h2>Reporter Module</h2>
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1. pLex6Op-pLEU2-NanoLuc [https://parts.igem.org/Part:BBa_K5477031| BBa_K5477031]: The reporter module contains Lex6Op, a cassette of six LexA operator sequences that serve as the binding sites for the LexA DBD in the receptor module. When LexA-ERα(LBD) is activated by estrogen binding, the LexA DBD binds to these operator sequences, activating the pLEU2 promoter. The pLEU2 promoter then drives the expression of the NanoLuc reporter gene. NanoLuc is a highly sensitive luciferase enzyme that produces bioluminescence when its substrate is added, allowing the system to output a quantifiable luminescent signal that directly correlates with the presence of estrogen or BPA.
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===Results===
  
 
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<partinfo>BBa_K5477043 parameters</partinfo>
 
<partinfo>BBa_K5477043 parameters</partinfo>
 
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===References===

Revision as of 11:05, 27 September 2024


Biosensor device I for detection of BPA

Summary

In this system, the pRET2-LexA-ERα(LBD) receptor module responds to the presence of estrogen or estrogen-like compounds, in this case BPA. When these ligands bind to the ERα LBD, it triggers a transcriptional response by allowing the LexA-ERα chimeric activator to bind to the Lex6Op sequences in the pLex6Op-pLEU2-NanoLuc reporter module. This activates the pLEU2 promoter, leading to the expression of NanoLuc, which produces a luminescent signal. The intensity of the luminescence is proportional to the concentration of the estrogenic compound in the environment, providing a measurable output of receptor activation.

Usage and Biology

In this biosensor system, the combination of receptor and reporter modules is designed to monitor the activity of Estrogen Receptor Alpha (ERα) in response to estrogenic compounds, such as estrogen or bisphenol A (BPA), and generate a luminescent signal through the expression of NanoLuc.

Why do we want to detect BPA in breast milk?

Receptor Module

1.pRET2-LexA-ERα(LBD) BBa_K5477027: This receptor module utilizes the pRET2 promoter to drive the expression of a fusion protein composed of the LexA DNA-binding domain (DBD) and the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ERα). The LexA-ERα(LBD) fusion allows the system to detect estrogen or estrogen-like molecules such as BPA. When BPA binds to the ERα LBD, it induces a conformational change in the receptor, activating the LexA DBD to bind to specific LexA operator sequences (Lex6Op).

Reporter Module

1. pLex6Op-pLEU2-NanoLuc BBa_K5477031: The reporter module contains Lex6Op, a cassette of six LexA operator sequences that serve as the binding sites for the LexA DBD in the receptor module. When LexA-ERα(LBD) is activated by estrogen binding, the LexA DBD binds to these operator sequences, activating the pLEU2 promoter. The pLEU2 promoter then drives the expression of the NanoLuc reporter gene. NanoLuc is a highly sensitive luciferase enzyme that produces bioluminescence when its substrate is added, allowing the system to output a quantifiable luminescent signal that directly correlates with the presence of estrogen or BPA.


Results

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1606
    Illegal PstI site found at 1790
    Illegal PstI site found at 1961
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 182
    Illegal PstI site found at 1790
    Illegal PstI site found at 1961
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1740
    Illegal BglII site found at 2751
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1606
    Illegal PstI site found at 1790
    Illegal PstI site found at 1961
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1606
    Illegal PstI site found at 1790
    Illegal PstI site found at 1961
    Illegal AgeI site found at 37
  • 1000
    COMPATIBLE WITH RFC[1000]


References