Difference between revisions of "Part:BBa K3734011"

 
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ELK is a kind of protein that can activate downstream gene expression after being regulated by phosphorylation.
 
ELK is a kind of protein that can activate downstream gene expression after being regulated by phosphorylation.
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                ELK
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                <p>ELK is generally dormant, when INSR accepts insulin to activate the MAPK pathway, Elk is phosphated into an active state, which can activate the expression of the gene downstream TRE.
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                1.Pattern diagram
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                    <img width="400px" src="https://2021.igem.org/wiki/images/d/d1/T--CSU_CHINA--tupian13.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 The model diagram TetR-ELK</p>
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                2.Experiments
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                2.1 Method
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                <p>We used Western Blot to detect phosphorylation of ERK1/ERK2 in the phosphorylation pathway with ERK antibodies.
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                2.2 Results
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                <img class="image" src="https://2021.igem.org/wiki/images/1/1f/T--CSU_CHINA--tupian16.png">
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                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.2 ERK phosphorylation changes with different insulin treatment</p>
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                <img class="image" src="https://2021.igem.org/wiki/images/b/b3/T--CSU_CHINA--tupian17.png">
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                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.3 ERK phosphorylation changes with different insulin treatment</p>
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                3.Caution
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                <p>Despite the length and the complication of phosphorylation pathway, the phosphorylation pathway of protein is a very short process, it is usually completed within minutes even seconds. The time of cracking the cells and collecting protein
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                    must be controlled precisely when detecting.
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                Reference：
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                <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.
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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===

Revision as of 10:08, 20 October 2021


ELK

ELK is a kind of protein that can activate downstream gene expression after being regulated by phosphorylation.

ELK

ELK is generally dormant, when INSR accepts insulin to activate the MAPK pathway, Elk is phosphated into an active state, which can activate the expression of the gene downstream TRE.

1.Pattern diagram

Fig.1 The model diagram TetR-ELK

2.Experiments

2.1 Method

We used Western Blot to detect phosphorylation of ERK1/ERK2 in the phosphorylation pathway with ERK antibodies.

2.2 Results

Fig.2 ERK phosphorylation changes with different insulin treatment

Fig.3 ERK phosphorylation changes with different insulin treatment

3.Caution

Despite the length and the complication of phosphorylation pathway, the phosphorylation pathway of protein is a very short process, it is usually completed within minutes even seconds. The time of cracking the cells and collecting protein must be controlled precisely when detecting.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 124
  • 1000
    COMPATIBLE WITH RFC[1000]