Part:BBa_K2368011
Introduction
Flag+T1R3 overlap
This part sweetness receptor T1R3 fusion with the Flag tag, just like the picture showed below.
Fig. 1 The schematic diagram of T1R3-Flag
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 The schematic diagram of making T1R3-Flag
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]
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