Difference between revisions of "Part:BBa K2368020"
 
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<h1>Introduction</h1>
 
<h1>Introduction</h1>
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<partinfo>BBa_K2368020 short</partinfo>
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<p style="text-align: center"><partinfo>BBa_K2368020 short</partinfo></p>
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<p> This part is the overlap of that sweetness receptor T1R3 combines with the BCP tag.  </p>
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<p> The T1R3 belongs to the C-class family of G protein coupling receptor (GPCR). And the C-terminal of GPCR that interacts with the G protein α subunit , which is a crucial factor during signal deliver through G protein signal pathway. Also, it is necessary for us to show whether the T1R3 expressed correctly. As a result of that, we can perform the fusion between the N-terminal of T1R3 and BCP tag.</P>
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<P> The result of this part is that the expression of the yellow cyanidin protein to detect the expression and the location of the T1R3.</p>
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<h1>Design</h1>
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<p> Firstly, we constructed the specific primers that consists of the overlap regions of the T1R3 and BCP. Then, we combined the overlap of T1R3 with the BCP by PCR.</p>
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[[File:T-BIT-China-2017parts-30.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 1 The schematic diagram of BCP+T1R3 overlap</p>
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[[File:T-BIT-China-2017parts-31.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 2 The PCR method of BCP+T1R3 overlap</p>
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<h1>Experiment</h1>
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<p>This part is used to detect the expression and the location of the T1R3.</p>
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[[File:T-BIT-China-2017parts-32.png|center|500px|默认文字]]
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<p style="text-align: center">Fig. 3 Electrophoresis of BCP+T1R3 overlap.Line two is the target. </p>
  
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<p>In order to confirm the expression and location of sweetness receptor, T1R2 and T1R3, we decide to construct the fusion protein, T1R3- amilCP (K592010). The part K592009 is blue chromoprotein and can exhibit strong blue color when expressed. Because the T1R3 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 T1R3 and C-terminal of amilCP. And hope to show the location of T1R3 through fluorescence microscope for detecting amilCP.</p>
 
  
 
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Latest revision as of 16:03, 28 October 2017


Introduction

BCP+T1R3 overlap

This part is the overlap of that sweetness receptor T1R3 combines with the BCP tag.

The T1R3 belongs to the C-class family of G protein coupling receptor (GPCR). And the C-terminal of GPCR that interacts with the G protein α subunit , which is a crucial factor during signal deliver through G protein signal pathway. Also, it is necessary for us to show whether the T1R3 expressed correctly. As a result of that, we can perform the fusion between the N-terminal of T1R3 and BCP tag.

The result of this part is that the expression of the yellow cyanidin protein to detect the expression and the location of the T1R3.

Design

Firstly, we constructed the specific primers that consists of the overlap regions of the T1R3 and BCP. Then, we combined the overlap of T1R3 with the BCP by PCR.

默认文字

Fig. 1 The schematic diagram of BCP+T1R3 overlap

默认文字

Fig. 2 The PCR method of BCP+T1R3 overlap

Experiment

This part is used to detect the expression and the location of the T1R3.

默认文字

Fig. 3 Electrophoresis of BCP+T1R3 overlap.Line two is the target.


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]