Translational_Unit

Part:BBa_K2207019

Designed by: Shisheng Li   Group: iGEM17_ZJU-China   (2017-10-21)


TRPV1-antiEGFP-2A-EGFP-Ferritin light chain-flag-Ferritin heavy chain

TRPV1 is fused with the antibody of EGFP while EGFP is fused with Ferritin. These two chimeric protein are co-expressed using 2A sequence which will separate the long peptide into two independent proteins with the help of ribosome. As for Ferritin, its light chain and heavy chain are linked by a flag tag which is recognized by corresponding antibody.

Lacking in the device which can generate the radio frequencies of proper power and frequency, we can only use heat shock and capsaicin to test this system. We use Fluo 4-AM to indicate the concentration change of calcium when the our yeast suffers a heat shock or be treated with capsaicin. The result shows that our BY4741 transferred with TRPV1 can be sensed a higher calcium concentration than the control when it has been heated or treated by capsaicin. As Fig. indicated, when the transgenic BY4741 is heated, the relative intracellular calcium ion concentration increases 71.6% while it increases 61.9% when treated with capsaicin.

ZJU_China_MWF_fig9.jpeg

This control had not been heated or treated with capsaicin.

We also made another control group which was treated with capsaicin.

ZJU_China_temp_Rplot06.jpeg


As the result shows, when treated by capsaicin, the transgenic BY4741 shows a higher intracellular calcium concentration than the untransgenic group, for about 24% enhancement. The difference is not as high as the former one, which suggests there may be some calcium channel similar with TRPV1 that enhanced the basic level of intracellular calcium concentration.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 787
    Illegal PstI site found at 1183
    Illegal PstI site found at 1202
    Illegal PstI site found at 1870
    Illegal PstI site found at 2476
    Illegal PstI site found at 2557
    Illegal PstI site found at 4519
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 394
    Illegal NheI site found at 1402
    Illegal NheI site found at 2956
    Illegal PstI site found at 787
    Illegal PstI site found at 1183
    Illegal PstI site found at 1202
    Illegal PstI site found at 1870
    Illegal PstI site found at 2476
    Illegal PstI site found at 2557
    Illegal PstI site found at 4519
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1313
    Illegal BglII site found at 1814
    Illegal BglII site found at 2950
    Illegal BamHI site found at 2836
    Illegal XhoI site found at 2783
    Illegal XhoI site found at 3886
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 787
    Illegal PstI site found at 1183
    Illegal PstI site found at 1202
    Illegal PstI site found at 1870
    Illegal PstI site found at 2476
    Illegal PstI site found at 2557
    Illegal PstI site found at 4519
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 787
    Illegal PstI site found at 1183
    Illegal PstI site found at 1202
    Illegal PstI site found at 1870
    Illegal PstI site found at 2476
    Illegal PstI site found at 2557
    Illegal PstI site found at 4519
    Illegal AgeI site found at 2594
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 53
    Illegal BsaI site found at 4190
    Illegal BsaI.rc site found at 461
    Illegal BsaI.rc site found at 1594
    Illegal BsaI.rc site found at 2105
    Illegal BsaI.rc site found at 2431
    Illegal SapI.rc site found at 2508


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