Difference between revisions of "Part:BBa K2368023"
 
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<h1>Introduction</h1>
 
<h1>Introduction</h1>
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<partinfo>BBa_K2368023 short</partinfo>
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<p style="text-align: center"><partinfo>BBa_K2368023 short</partinfo></p>
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<p> This part sweetness receptor T1R2 fusion with the Myc tag, just like the picture showed below. </p>
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[[File:T-BIT-China-2017parts-40.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 1 The schematic diagram of Myc+T1R2 overlap</p>
  
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<p>In order to confirm the expression and location of sweetness receptor, T1R2 and T1R3, we decide to construct T1R2-Myc tag. This tag can be recognized by specific monoclonal antibodies then we use fluorescence second antibody to interact with monoclonal antibodies to show the T1R2’s expression and location. </p>
 
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<p>Because the T1R2 is belonged to C-class family G protein coupling receptor (GPCR) and the carboxyl terminal of GPCR interacts with the G protein αsubunit ,which is a crucial factor during signal deliver through G protein signal pathway, we perform the fusion between the N-terminal of T1R2 and Myc tag. And hope to show the location of T1R2 through immunofluorescence for detecting Myc tag.</p>
 
  
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<h1>Design</h1>
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<p> In order to show whether the sweet receptor T1R2 are translated and located correctly, we fusion the Myc tag to N-terminal of T1R2. The reason fused tag to N-terminal is that the C-terminal of T1R2 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 T1R2 is located at the right position. </p>
  
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[[File:T-BIT-China-2017parts-41.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 2 The schematic diagram of making Myc+T1R2 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 Myc tag. Then, fused T1R2 with the Myc using PCR. The length of sequence is 349 bp. </p>
 
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===Usage and Biology===
 
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<h2>Sequence and Features</h2>
 
<h2>Sequence and Features</h2>
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<partinfo>BBa_K2368023 SequenceAndFeatures</partinfo>
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===Functional Parameters===
 
===Functional Parameters===
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Latest revision as of 16:04, 28 October 2017


Introduction

Myc+T1R2 overlap

This part sweetness receptor T1R2 fusion with the Myc tag, just like the picture showed below.

默认文字

Fig. 1 The schematic diagram of Myc+T1R2 overlap


Design

In order to show whether the sweet receptor T1R2 are translated and located correctly, we fusion the Myc tag to N-terminal of T1R2. The reason fused tag to N-terminal is that the C-terminal of T1R2 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 T1R2 is located at the right position.


默认文字

Fig. 2 The schematic diagram of making Myc+T1R2 overlap

Experiment

At the beginning, we construct the specific primer that consists of the gene sequence of Myc tag. Then, fused T1R2 with the Myc using PCR. The length of sequence is 349 bp.

Sequence and Features


Assembly Compatibility:
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    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
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    COMPATIBLE WITH RFC[21]
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    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]