Difference between revisions of "Part:BBa K5107002"

 
 
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<partinfo>BBa_K5107002 short</partinfo>
 
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HRE5 is the 5 times repeats of th HRE minimal to increase the interaction surface of the hormone receptor with the DNA.
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HRE5 is the five tandem repeats of the HRE minimal:([https://parts.igem.org/Part:BBa_K5107000 BBa_K5107000]) to increase the interaction surface of the hormone receptor with the DNA.
  
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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Previously, in-cell systems have successfully used five tandem repeats of the REs for different human hormonal receptors<html><a href="#ref1">[1]</a></html>.
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Inserted in plasmids such as those we used for our in cell-assay, it is shown efficient to express Lac-Z when the associated hormone-receptor complex is present in the cell. In this manner, hormone work as trancription activator.
  
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K5107002 SequenceAndFeatures</partinfo>
 
  
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<center>
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    <figure>
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        <img src="https://static.igem.wiki/teams/5107/eng-incellsystem.webp"width="500" height="500">
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        <figcaption><b>Figure 1:</b>Schematic overview of the in cell system which uses the Lac-Z as reporter gene when hormone/edc i spresent in the cell.</figcaption>
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    </figure>
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</center>
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</html>
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===Assemply===
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*These elements are part of the plasmid fist presented by (Miller et all,2010)<html><a href="#ref1">[1]</a></html>. For the list of the plasmid check [https://parts.igem.org/Part:BBa_K5107000 BBa_K5107000].
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*We used specific primers to amplify the REs from two of these plasmids(<strong>pRR-ERalpha-5Z,pRR-MR-5Z</strong>). The plamsids were a gift from Charles Miller (Addgene plasmid # 23061, # 23059 ; http://n2t.net/addgene:23061,http://n2t.net/addgene:23059  ; RRID:Addgene_23061,RRID:Addgene_23059).
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{| class="wikitable"
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|+ Primers for HRE5 and ERE5
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|-
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! !! Forward Primer !! Reverse Primer
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|-
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| HRE5 || ataggtagaNCCGTCTGGTACAGGGTGTTCTT || atgtgggNCCCCGGGAAAAAGAACACCCTGTA
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|-
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| ERE5 || ataggtagaNCCCAGGTCAGAGTGACCTG || atgtgggNCCGACCCCGGGAAAAAGC
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|}
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*After that, USER Cloning was performed so the final constucts are derived. [https://parts.igem.org/Part:BBa_K5107006 BBa_K5107006] and [https://parts.igem.org/Part:BBa_K5107012 BBa_K5107012]
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===Sequence and Features===
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<partinfo>BBa_K5107002 SequenceandFeatures</partinfo>
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K5107002 parameters</partinfo>
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<partinfo>BBa_K5107000 parameters</partinfo>
 
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===References===
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<html>
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<ol>
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    <li id="ref1">
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        Miller, C. A., Tan, X., Wilson, M., Bhattacharyya, S., & Ludwig, S. (2010). 
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        Single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays. <i>Plasmid</i>, <i>63</i>(2), 73–78. https://doi.org/10.1016/j.plasmid.2009.11.003
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Latest revision as of 09:14, 30 September 2024


HRE5


HRE5 is the five tandem repeats of the HRE minimal:(BBa_K5107000) to increase the interaction surface of the hormone receptor with the DNA.


Usage and Biology

Previously, in-cell systems have successfully used five tandem repeats of the REs for different human hormonal receptors[1]. Inserted in plasmids such as those we used for our in cell-assay, it is shown efficient to express Lac-Z when the associated hormone-receptor complex is present in the cell. In this manner, hormone work as trancription activator.


Figure 1:Schematic overview of the in cell system which uses the Lac-Z as reporter gene when hormone/edc i spresent in the cell.

Assemply

  • These elements are part of the plasmid fist presented by (Miller et all,2010)[1]. For the list of the plasmid check BBa_K5107000.
  • We used specific primers to amplify the REs from two of these plasmids(pRR-ERalpha-5Z,pRR-MR-5Z). The plamsids were a gift from Charles Miller (Addgene plasmid # 23061, # 23059 ; http://n2t.net/addgene:23061,http://n2t.net/addgene:23059  ; RRID:Addgene_23061,RRID:Addgene_23059).


Primers for HRE5 and ERE5
Forward Primer Reverse Primer
HRE5 ataggtagaNCCGTCTGGTACAGGGTGTTCTT atgtgggNCCCCGGGAAAAAGAACACCCTGTA
ERE5 ataggtagaNCCCAGGTCAGAGTGACCTG atgtgggNCCGACCCCGGGAAAAAGC


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



References

  1. Miller, C. A., Tan, X., Wilson, M., Bhattacharyya, S., & Ludwig, S. (2010). Single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays. Plasmid, 63(2), 73–78. https://doi.org/10.1016/j.plasmid.2009.11.003