Difference between revisions of "Part:BBa K2368009"
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| − | [[File:T-BIT-China- | + | [[File:T-BIT-China-2017yhy-111.png|center|500px|默认文字]] |
| − | <p style="text-align: center"> | + | <p style="text-align: center">Fig.1 The schematic diagram of making T1R2-His</p> |
<h1>Experiment</h1> | <h1>Experiment</h1> | ||
<p> 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. </p> | <p> 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. </p> | ||
Revision as of 09:11, 28 October 2017
Introduction
His+T1R2 overlap
This part sweetness receptor T1R2 fusion with the His tag, just like the picture showed below.
fig.1 Fig.1 The schematic diagram of T1R2-His
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.
Fig.1 The schematic diagram of making T1R2-His
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]