Difference between revisions of "Part:BBa K2368011"

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__NOTOC__
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<h1>Introduction</h1>
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<p style="text-align: center"><partinfo>BBa_K2368011 short</partinfo></p>
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<p> This part sweetness receptor T1R3 fusion with the Flag tag, just like the picture showed below. </p>
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[[File:T-BIT-China-2017parts-45.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 1 The schematic diagram of T1R3-Flag</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 Flag 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-46.png|center|500px|默认文字]]
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<p style="text-align: center"><span>Fig. 2 The schematic diagram of making T1R3-Flag</span></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 Flag tag. Then, fused T1R3 with the Flag using PCR. The length of sequence is 50bp. </p>
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===Usage and Biology===
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<h2>Sequence and Features</h2>
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<partinfo>BBa_K2368011 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K2368011 parameters</partinfo>
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Latest revision as of 16:00, 28 October 2017


Introduction

Flag+T1R3 overlap

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

默认文字

Fig. 1 The schematic diagram of T1R3-Flag


Design

In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the Flag 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 T1R3-Flag

Experiment

At the beginning, we construct the specific primer that consists of the gene sequence of Flag tag. Then, fused T1R3 with the Flag using PCR. The length of sequence is 50bp.

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]