Difference between revisions of "Part:BBa K5477023"
 
(4 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K5477023 short</partinfo>
 
<partinfo>BBa_K5477023 short</partinfo>
  
−
&lt;a href="https://parts.igem.org/Part:BBa_K3793004"&gt;BBa_K3793004&lt;/a&gt;
+
The pSTE12-AhR receptor module is a composite designed to harness the natural ligand-sensing ability of the Aryl Hydrocarbon Receptor (AhR), integrated with the yeast promoter pSTE12 [https://parts.igem.org/Part:BBa_K5477001 |BBa_K5477001]. In this module, AhR [https://parts.igem.org/Part:BBa_K3793004| BBa_K3793004]is expressed under the control of the pSTE12 promoter. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.
  
  
−
The pSTE12-AhR receptor module is a composite designed to harness the natural ligand-sensing ability of the Aryl Hydrocarbon Receptor (AhR), integrated with the yeast promoter pSTE12 &lt;a href="https://parts.igem.org/Part:BBa_K5477001"&gt;BBa_K5477001&lt;/a&gt;. In this module, AhR &lt;a href="https://parts.igem.org/Part:BBa_K3793004"&gt;BBa_K3793004&lt;/a&gt; is expressed under the control of the pSTE12 promoter. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.
+
In our pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequence in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device:  
−
 
+
−
 
+
−
In our pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequences in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device:  
+
  
  

Latest revision as of 19:20, 26 September 2024


pSTE12-AhR - receptor module

The pSTE12-AhR receptor module is a composite designed to harness the natural ligand-sensing ability of the Aryl Hydrocarbon Receptor (AhR), integrated with the yeast promoter pSTE12 |BBa_K5477001. In this module, AhR BBa_K3793004is expressed under the control of the pSTE12 promoter. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.


In our pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequence in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device:


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 927
    Illegal NheI site found at 732
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 927
    Illegal XbaI site found at 55
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 262
    Illegal BsaI.rc site found at 443


References

1. Carambia, A., Schuran, F.A. The aryl hydrocarbon receptor in liver inflammation. Semin Immunopathol 43, 563–575 (2021). https://doi.org/10.1007/s00281-021-00867-8

2. Mandal A, Biswas N, Alam MN. Implications of xenobiotic-response element(s) and aryl hydrocarbon receptor in health and diseases. Hum Cell. 2023 Sep;36(5):1638-1655. doi: 10.1007/s13577-023-00931-5. Epub 2023 Jun 17. PMID: 37329424.