Difference between revisions of "Part:BBa K2368009"

Line 6: Line 6:
 
]
 
]
 
[[File:T-BIT-China-2017parts-38.png|center|500px|默认文字]]
 
[[File:T-BIT-China-2017parts-38.png|center|500px|默认文字]]
<p style="text-align: center">Fig.1 The schematic diagram of HIS+T1R2 overlap</p>
+
 
  
  
Line 14: Line 14:
  
 
[[File:T-BIT-China-2017parts-39.png|center|500px|默认文字]]
 
[[File:T-BIT-China-2017parts-39.png|center|500px|默认文字]]
<p style="text-align: center">Fig.2 Electrophoresis of HIS+T1R2 overlap. </p>
 
  
 
<h1>Experiment</h1>
 
<h1>Experiment</h1>

Revision as of 11:12, 26 October 2017


Introduction

His+T1R2 overlap

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

]

默认文字


Design

In order to show whether the sweet receptor T1R2 are translated and located correctly, we fusion the HIS 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

Experiment

At the beginning, we construct the specific primer that consists of the gene sequence of HIS tag. Then, fused T1R2 with the HIS 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]