Difference between revisions of "Part:BBa K2368024"

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__NOTOC__
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<h1>Introduction</h1>
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<p style="text-align: center"><partinfo>BBa_K2368024 short</partinfo></p>
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<p> This part sweetness receptor T1R3 fusion with the His tag, just like the picture showed below. </p>
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[[File:T-BIT-China-2017parts-42.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 1 The schematic diagram of His+T1R3 overlap</p>
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<h1>Design</h1>
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<p> In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the His tag to N-terminal of T1R3. The reason fused tag to N-terminal is that the C-terminal of T1R3 interacts with the G protein α subunit, which is a crucial factor during signal deliver through G protein signal pathway. And the N-terminal is outside the cell, so it can be detected without killing it, using immunofluorescence can figure out whether T1R3 is located at the right position. </p>
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[[File:T-BIT-China-2017parts-43.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 2 The schematic diagram of making His+T1R3 overlap</p>
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<h1>Experiment</h1>
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<p> At the beginning, we construct the specific primer that consists of the gene sequence of His tag. Then, fused T1R3 with the HIS using PCR. The length of sequence is 187bp. </p>
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<h2>Sequence and Features</h2>
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<partinfo>BBa_K2368024 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_K2368024 parameters</partinfo>
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Latest revision as of 16:02, 28 October 2017


Introduction

His+T1R3 overlap

This part sweetness receptor T1R3 fusion with the His tag, just like the picture showed below.

默认文字

Fig. 1 The schematic diagram of His+T1R3 overlap


Design

In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the His tag to N-terminal of T1R3. The reason fused tag to N-terminal is that the C-terminal of T1R3 interacts with the G protein α subunit, which is a crucial factor during signal deliver through G protein signal pathway. And the N-terminal is outside the cell, so it can be detected without killing it, using immunofluorescence can figure out whether T1R3 is located at the right position.


默认文字

Fig. 2 The schematic diagram of making His+T1R3 overlap


Experiment

At the beginning, we construct the specific primer that consists of the gene sequence of His tag. Then, fused T1R3 with the HIS using PCR. The length of sequence is 187bp.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 12
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 95
  • 1000
    COMPATIBLE WITH RFC[1000]