Part:BBa_K2368024
Introduction
His+T1R3 overlap
This part sweetness receptor T1R3 fusion with the His tag, just like the picture showed below.
Fig.1 The schematic diagram of His+T1R3 overlap
Design
In order to show whether the sweet receptor T1R3 are translated and located correctly, we fusion the His 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 His+T1R3 overlap
Experiment
At the beginning, we construct the specific primer that consists of the gene sequence of His tag. Then, fused T1R3 with the HIS using PCR. The length of sequence is 187bp.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 12
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 95
- 1000COMPATIBLE WITH RFC[1000]
| None |