Coding

Part:BBa_K3852005

Designed by: Yuan Cao   Group: iGEM21_BIT-China   (2021-10-01)
Revision as of 11:05, 17 October 2021 by Caoyuan (Talk | contribs) (→‎Experiment)


T1R1

The protein encoded by this gene is a G protein-coupled receptor and is a component of the heterodimeric amino acid taste receptor T1R1+3. The T1R1+3 receptor responds to L-amino acids but not to D-enantiomers or other compounds. Most amino acids that are perceived as sweet activate T1R1+3, and this activation is strictly dependent on an intact T1R1+3 heterodimer. Multiple transcript variants encoding different isoforms have been found for this gene.

T1R1 is essentially a recognition receptor for the nitrogen signal in the body, and it may recognize the umami substance through its amino group.In our experiments, we used this gene to express umami taste receptors in Saccharomyces cerevisiae.

Usage and Biology

Putative taste receptor. TAS1R1/TAS1R3 responds to the umami taste stimulus (the taste of monosodium glutamate). Sequence differences within and between species can significantly influence the selectivity and specificity of taste responses.We acquire it from synthesis company.

Experiment

After literature research, we found that the taste receptors in the human body are made up of T1R1 and T1R3.So we plan to introduce T1R1 and T1R3 into the Saccharomyces cerevisiae. The first step is to integrate the two genes into pESC-LEU, and then use semi-lactose induction to allow the two genes to be expressed in the Saccharomyces cerevisiae at the same time.

The design of the plasmid is shown below:

Figure1. pESC-LEU with T1R1 and T1R3
Figure2. Design diagram (where SS is Ste2 signal peptide sequence)
Figure3. Running glue picture after PCR (right-most strip is T1R1)
Figure4. A colony that grows after transferring plasmids that express umami taste receptors

Reference

[1] Huang Yulin, Lu Dingqiang, Liu Hai et al. Preliminary research on the receptor-ligand recognition mechanism of umami by an hT1R1 biosensor[J] Food & Function.2019,10(3): 1280-1287.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2491
    Illegal XbaI site found at 15
    Illegal XbaI site found at 854
    Illegal XbaI site found at 944
    Illegal XbaI site found at 1262
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2491
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2491
    Illegal BglII site found at 218
    Illegal BglII site found at 1820
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2491
    Illegal XbaI site found at 15
    Illegal XbaI site found at 854
    Illegal XbaI site found at 944
    Illegal XbaI site found at 1262
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2491
    Illegal XbaI site found at 15
    Illegal XbaI site found at 854
    Illegal XbaI site found at 944
    Illegal XbaI site found at 1262
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1440


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