Difference between revisions of "Part:BBa K2368011"
| Line 2: | Line 2: | ||
__NOTOC__ | __NOTOC__ | ||
<h1>Introduction</h1> | <h1>Introduction</h1> | ||
| − | <partinfo> | + | <partinfo>BBa_K2368024 short</partinfo> |
<p> This part sweetness receptor T1R3 fusion with the FLAG tag, just like the picture showed below. </p> | <p> This part sweetness receptor T1R3 fusion with the FLAG tag, just like the picture showed below. </p> | ||
| − | + | ||
| − | <p> Fig.1 The schematic diagram of FLAG+T1R3 overlap </p> | + | [[File:T-BIT-China-2017parts-45.png|center|500px|默认文字]] |
| + | <p style="text-align: center">Fig.1 The schematic diagram of FLAG+T1R3 overlap</p> | ||
| + | |||
| + | |||
<h1>Design</h1> | <h1>Design</h1> | ||
| − | <p> In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the FLAG tag to N-terminal of T1R3. The reason fused tag to N-terminal is that the C-terminal of T1R3 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 T1R3 is located at the right position.</p> | + | <p> In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the FLAG tag to N-terminal of T1R3. The reason fused tag to N-terminal is that the C-terminal of T1R3 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 T1R3 is located at the right position. </p> |
| − | + | ||
| − | <p> Fig.2 Electrophoresis of FLAG+T1R3 overlap.</p> | + | |
| + | [[File:T-BIT-China-2017parts-46.png|center|500px|默认文字]] | ||
| + | <p style="text-align: center">Fig.2 Electrophoresis of FLAG+T1R3 overlap. </p> | ||
| + | |||
<h1>Experiment</h1> | <h1>Experiment</h1> | ||
<p> At the beginning, we construct the specific primer that consists of the gene sequence of FLAG tag. Then, fused T1R3 with the FLAG 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 FLAG tag. Then, fused T1R3 with the FLAG using PCR. The length of sequence is 50bp. </p> | ||
| Line 17: | Line 23: | ||
<!-- --> | <!-- --> | ||
<h2>Sequence and Features</h2> | <h2>Sequence and Features</h2> | ||
| − | <partinfo> | + | <partinfo>BBa_K2368009 SequenceAndFeatures</partinfo> |
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
| − | <partinfo> | + | <partinfo>BBa_K2368009 parameters</partinfo> |
<!-- --> | <!-- --> | ||
Revision as of 18:24, 25 October 2017
Introduction
His+T1R3 overlap
This part sweetness receptor T1R3 fusion with the FLAG tag, just like the picture showed below.
Fig.1 The schematic diagram of FLAG+T1R3 overlap
Design
In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the FLAG tag to N-terminal of T1R3. The reason fused tag to N-terminal is that the C-terminal of T1R3 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 T1R3 is located at the right position.
Fig.2 Electrophoresis of FLAG+T1R3 overlap.
Experiment
At the beginning, we construct the specific primer that consists of the gene sequence of FLAG tag. Then, fused T1R3 with the FLAG 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]