Difference between revisions of "Part:BBa K3734009"

 
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<partinfo>BBa_K3734009 short</partinfo>
  
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                INSR
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                <p>Insulin receptor  (INSR)is a membrane protein that bid to insulin, feel insullin concentration and activate the downstream MAPK phosphorylation pathway. We have designed a carrier CMV-INSR-EF1A-ZsGreen containing INSR to express INSR, and its expression can be characterized by the green fluorescent protein ZsGreen. Experiments have proved that INSR is successfully expressed in our engineering cells.
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                1.Experiment
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                1.1 Method
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                <p>Observation of green fluorescent protein ZsGreen under laser confocal microscope.
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                1.2 Result
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                    <img width="400px" src="https://2021.igem.org/wiki/images/a/a7/T--CSU_CHINA--tupian11.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 Electrophoretic diagram of INSR PCR product</p>
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                <p>INSR is successfully expressed and working in cell 293T
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                    <img width="400px" src="https://2021.igem.org/wiki/images/e/ef/T--CSU_CHINA--tupian12.png"></p>
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                <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.2 Green fluorescence of INSR expression</p>
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                2.Caution
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                <p>Because the INSR sequence is very long, it is difficult for DH5α receptor cells containing recombinant enzymes to meet the requirements during cloning, and receptor cells of recombinant enzymes may need to be removed.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3734009 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K3734009 parameters</partinfo>
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Revision as of 14:35, 20 October 2021

INSR


INSR

Insulin receptor  (INSR)is a membrane protein that bid to insulin, feel insullin concentration and activate the downstream MAPK phosphorylation pathway. We have designed a carrier CMV-INSR-EF1A-ZsGreen containing INSR to express INSR, and its expression can be characterized by the green fluorescent protein ZsGreen. Experiments have proved that INSR is successfully expressed in our engineering cells.

1.Experiment

1.1 Method

Observation of green fluorescent protein ZsGreen under laser confocal microscope.

1.2 Result

Fig.1 Electrophoretic diagram of INSR PCR product

INSR is successfully expressed and working in cell 293T

Fig.2 Green fluorescence of INSR expression

2.Caution

Because the INSR sequence is very long, it is difficult for DH5α receptor cells containing recombinant enzymes to meet the requirements during cloning, and receptor cells of recombinant enzymes may need to be removed.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 551
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2055
    Illegal BglII site found at 8438
    Illegal BamHI site found at 2401
    Illegal XhoI site found at 809
    Illegal XhoI site found at 2566
    Illegal XhoI site found at 3579
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 537
    Illegal NgoMIV site found at 2639
    Illegal AgeI site found at 7134
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2267
    Illegal BsaI site found at 2577
    Illegal BsaI.rc site found at 502
    Illegal BsaI.rc site found at 744
    Illegal BsaI.rc site found at 1319
    Illegal BsaI.rc site found at 1775
    Illegal BsaI.rc site found at 2097
    Illegal BsaI.rc site found at 3674
    Illegal BsaI.rc site found at 3849
    Illegal SapI site found at 790
    Illegal SapI site found at 8692
    Illegal SapI.rc site found at 3856
    Illegal SapI.rc site found at 9122