Difference between revisions of "Part:BBa K2368015"
 
(7 intermediate revisions by 2 users not shown)
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<h1>Introduction</h1>
 
<h1>Introduction</h1>
−
<partinfo>BBa_K2368015 short</partinfo>
+
<p style="text-align: center"><partinfo>BBa_K2368015 short</partinfo></p>
 +
<p>This part is the overlap of that sweetness receptor T1R2 combines with the GFP tag. </p>
 +
<p> The T1R2 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 T1R2 expressed correctly. As a result of that, we can perform the fusion between the N-terminal of T1R2 and GFP tag.</P>
 +
<P> The result of this part is that the expression of the yellow cyanidin protein to detect the expression and the location of the T1R2.</p>
 +
<h1>Design</h1>
 +
<p>Firstly, we constructed the specific primers that consists of the overlap regions of the T1R2 and GFP. Then, we combined the overlap of T1R3 with the GFP by PCR.</p>
 +
[[File:T-BIT-China-2017parts-27.png|center|500px|默认文字]]
 +
<p style="text-align: center">Fig.1 The schematic diagram of GFP+T1R2 overlap</p>
 +
[[File:T-BIT-China-2017parts-28.png|center|500px|默认文字]]
 +
<p style="text-align: center">Fig. 2 The PCR method of GFP+T1R2 overlap</p>
 +
 
 +
<h1>Experiment</h1>
 +
<p>This part is used to detect the expression and the location of the T1R2.</p>
 +
[[File:T-BIT-China-2017parts-29.png|center|300px|默认文字]]
 +
<p style="text-align: center">Fig.3 Electrophoresis of GFP+T1R2 overlap. </p>
  
−
<p>In order to confirm the expression and location of sweetness receptor, T1R2 and T1R3, we decide to construct the fusion protein T1R2-GFP (E0040). 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 &#945;subunit ,which is a crucial factor during signal deliver through G protein signal pathway, we perform the fusion between the N-terminal of T1R2 and C-terminal of GFP. And hope to show the location of T1R2 through fluorescence microscope for detecting GFP.</p>
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 10: Line 23:
  
 
<!-- -->
 
<!-- -->
−
<h2>Sequence and Features<h2>
+
<h2>Sequence and Features</h2>
 
<partinfo>BBa_K2368015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2368015 SequenceAndFeatures</partinfo>
  

Latest revision as of 19:11, 26 October 2017


Introduction

T1R2-GFP

This part is the overlap of that sweetness receptor T1R2 combines with the GFP tag.

The T1R2 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 T1R2 expressed correctly. As a result of that, we can perform the fusion between the N-terminal of T1R2 and GFP tag.

The result of this part is that the expression of the yellow cyanidin protein to detect the expression and the location of the T1R2.

Design

Firstly, we constructed the specific primers that consists of the overlap regions of the T1R2 and GFP. Then, we combined the overlap of T1R3 with the GFP by PCR.

默认文字

Fig.1 The schematic diagram of GFP+T1R2 overlap

默认文字

Fig. 2 The PCR method of GFP+T1R2 overlap

Experiment

This part is used to detect the expression and the location of the T1R2.

默认文字

Fig.3 Electrophoresis of GFP+T1R2 overlap.


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]