Revision as of 18:25, 25 October 2017

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 Electrophoresis of 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