Difference between revisions of "Part:BBa K2368011"

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[[File:T-BIT-China-2017parts-45.png|center|500px|默认文字]]
 
[[File:T-BIT-China-2017parts-45.png|center|500px|默认文字]]
<p style="text-align: center">Fig.1 The schematic diagram of FLAG+T1R3 overlap</p>
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[[File:T-BIT-China-2017parts-46.png|center|500px|默认文字]]
 
[[File:T-BIT-China-2017parts-46.png|center|500px|默认文字]]
<p style="text-align: center">Fig.2 Electrophoresis of FLAG+T1R3 overlap. </p>
 
  
 
<h1>Experiment</h1>
 
<h1>Experiment</h1>

Revision as of 11:16, 26 October 2017


Introduction

His+T1R3 overlap

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

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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.


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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]