Difference between revisions of "Part:BBa K3734012"

 
 
(2 intermediate revisions by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3734012 short</partinfo>
 
<partinfo>BBa_K3734012 short</partinfo>
  
 
TRE is a DNA structure domain that combined with TetR.
 
TRE is a DNA structure domain that combined with TetR.
−
 
+
<html>
 +
<head>
 +
           
 +
</head>
 +
       
 +
<body>
 +
<h2 class="pageContent-main__title">
 +
                <!--put tile here, <h2>title</h2>, class="pageContent-main__title" means it is the main title-->
 +
                TRE
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p>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 phosphated 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.
 +
                </p>
 +
                <!--put text here, <p>content</p>-->
 +
            </div>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                1.Pattern diagram
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p style="text-align: center;">
 +
                    <img width="400px" height="400" src="https://2021.igem.org/wiki/images/c/c0/T--CSU_CHINA--tupian18.png"></p>
 +
                <!--put image's url here-->
 +
                <br>
 +
                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The model diagram of TRE's work</p>
 +
            </div>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                2.Experiment
 +
            </h2>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                2.1 Method
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p>Phosphated ELK will activate the expression of downstream report gene mCherry after the combination of TRE and TetR. We used lase confocal microscope to observe the expression.
 +
                </p>
 +
                <!--put text here, <p>content</p>-->
 +
            </div>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                2.2 Results
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p>TRE works well.
 +
                </p>
 +
                <p style="text-align: center;">
 +
                    <img width="400px" src="https://2021.igem.org/wiki/images/5/54/T--CSU_CHINA--tupian14.png"></p>
 +
                <!--put image's url here-->
 +
                <br>
 +
                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.2 Under Laser confocal microscopy, fluorescence of mCherry expression downstream of Tet-Off system</p>
 +
            </div>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                3.Caution
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p>If the green fluorescent protein is expressed too much, and if the green fluorescence is too strong, it may cause a series of colors. In order to ensure the accuracy of the experimental results, the level of noise reduction should be raised.
 +
                </p>
 +
            </div>
 +
            <h2 class="pageContent-main__title pageContent-main__subtitle">
 +
                <!--class="pageContent-main__title" means it is the sub title-->
 +
                Reference：
 +
            </h2>
 +
            <div class="pageContent-main__textBox">
 +
                <!--all the content must included in this div-->
 +
                <p>[1]Haifeng Ye, Mingqi Xie, Shuai Xue.Self-adjusting synthetic gene circuit for correcting insulin resistance[J].Nat Biomed Eng.2017 Jan;1(1):0005.
 +
                </p>
 +
            </div>
 +
</body>
 +
</html>
 
<!-- 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 10:21, 20 October 2021

TRE

TRE is a DNA structure domain that combined with TetR.

TRE

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 phosphated 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.

1.Pattern diagram


Fig.1 The model diagram of TRE's work

2.Experiment

2.1 Method

Phosphated ELK will activate the expression of downstream report gene mCherry after the combination of TRE and TetR. We used lase confocal microscope to observe the expression.

2.2 Results

TRE works well.


Fig.2 Under Laser confocal microscopy, fluorescence of mCherry expression downstream of Tet-Off system

3.Caution

If the green fluorescent protein is expressed too much, and if the green fluorescence is too strong, it may cause a series of colors. In order to ensure the accuracy of the experimental results, the level of noise reduction should be raised.

Reference：

[1]Haifeng Ye, Mingqi Xie, Shuai Xue.Self-adjusting synthetic gene circuit for correcting insulin resistance[J].Nat Biomed Eng.2017 Jan;1(1):0005.

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]