Difference between revisions of "Part:BBa K364304"

 
 
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The Tetracycline Response Element (TRE) is recognized and bound by the Tetracycline repressor (TetR) protein. The TRE consists of 7 repeats separated by spacer sequences and placed upstream of CMV minimal promoter that has basal expession in the abscence of bound TetR. Tetracycline derivatives (e.g. doxycycline) bind TetR and render it incapable of binding to TRE, thereby forcing the expression of target genes.
 
The Tetracycline Response Element (TRE) is recognized and bound by the Tetracycline repressor (TetR) protein. The TRE consists of 7 repeats separated by spacer sequences and placed upstream of CMV minimal promoter that has basal expession in the abscence of bound TetR. Tetracycline derivatives (e.g. doxycycline) bind TetR and render it incapable of binding to TRE, thereby forcing the expression of target genes.
  
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[[Image:Team Debrecen TRE.jpg|800px|thumb|center|Picture of gel electrophoresis: TRE-CMV in pSB1C3 resulting a 321 bp long insert]]
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        Supplementary Experiment
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        <p>Group: CSU_CHINA 2021
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        <p>Author: Xingjun Zhao
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        <p>Summary: TRE is a DNA structure domain that combined with TetR. It is a part of Tet-off system. Our first purpose of using it is to make phosphorylated ELK1 to locate and activate the expression of downstream genes (miRNA). The second reason we
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            used it is because of the Tet-off system consists of TetR and TRE can be blocked by tetracycline. Tetracycline will stop the combination between TetR and TRE, stop the system from working. This provides a mandatory brake system for our loop.
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            We can make sure the security and prevent the hypoglycemia caused by abnormal feedback through exogenous tetracycline.
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            <img width="400px" src="https://2021.igem.org/wiki/images/1/1b/T--CSU_CHINA--pic6.png"></p>
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        <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 Under Laser confocal microscopy, fluorescence of ZsGreen and mCherry expression downstream of Tet-Off system</p>
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        <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.2 Transcriptional level analysis of downstream mCherry in TEt-OFF system without tetracycline</p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 18:44, 20 October 2021

TRE-CMV

The Tetracycline Response Element (TRE) is recognized and bound by the Tetracycline repressor (TetR) protein. The TRE consists of 7 repeats separated by spacer sequences and placed upstream of CMV minimal promoter that has basal expession in the abscence of bound TetR. Tetracycline derivatives (e.g. doxycycline) bind TetR and render it incapable of binding to TRE, thereby forcing the expression of target genes.

Picture of gel electrophoresis: TRE-CMV in pSB1C3 resulting a 321 bp long insert

Supplementary Experiment

Group: CSU_CHINA 2021

Author: Xingjun Zhao

Summary: TRE is a DNA structure domain that combined with TetR. It is a part of Tet-off system. Our first purpose of using it is to make phosphorylated ELK1 to locate and activate the expression of downstream genes (miRNA). The second reason we used it is because of the Tet-off system consists of TetR and TRE can be blocked by tetracycline. Tetracycline will stop the combination between TetR and TRE, stop the system from working. This provides a mandatory brake system for our loop. We can make sure the security and prevent the hypoglycemia caused by abnormal feedback through exogenous tetracycline.

Fig.1 Under Laser confocal microscopy, fluorescence of ZsGreen and mCherry expression downstream of Tet-Off system

Fig.2 Transcriptional level analysis of downstream mCherry in TEt-OFF system without tetracycline

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]