Difference between revisions of "Team:SUSTech Shenzhen/Part Collection"
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| − | Our | + | Our '''Part Collection''' includes: |
* Original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>); | * Original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>); | ||
* Five mutated TRPC5 plasmids(<partinfo>BBa_K1943022</partinfo>, <partinfo>BBa_K1943023</partinfo>, <partinfo>BBa_K1943024</partinfo>, <partinfo>BBa_K1943025</partinfo>, <partinfo>BBa_K1943026</partinfo>). | * Five mutated TRPC5 plasmids(<partinfo>BBa_K1943022</partinfo>, <partinfo>BBa_K1943023</partinfo>, <partinfo>BBa_K1943024</partinfo>, <partinfo>BBa_K1943025</partinfo>, <partinfo>BBa_K1943026</partinfo>). | ||
Transient receptor potential channel 5 (TRPC5) is a mechanosensitive (MS) channel[1] which takes charge of hearing in our project. Future teams could use original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>) to sense sound or other kinds of mechanical forces. Furthermore, we randomly mutated the ankyrin region of TRPC5 channel by PCR. We expected the ankyrin region to be a crucial part for sense mechanical forces based on the structure of TRPC5[2].(Detailed methods are shown in our notebook.) | Transient receptor potential channel 5 (TRPC5) is a mechanosensitive (MS) channel[1] which takes charge of hearing in our project. Future teams could use original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>) to sense sound or other kinds of mechanical forces. Furthermore, we randomly mutated the ankyrin region of TRPC5 channel by PCR. We expected the ankyrin region to be a crucial part for sense mechanical forces based on the structure of TRPC5[2].(Detailed methods are shown in our notebook.) | ||
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<html><a href="/ Team:SUSTech_Shenzhen/Notebook/Molecular" class="btn btn-success">Learn More</a></html> | <html><a href="/ Team:SUSTech_Shenzhen/Notebook/Molecular" class="btn btn-success">Learn More</a></html> | ||
Revision as of 06:25, 19 October 2016
Part Collection
Contribution
Our Part Collection includes:
- Original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>);
- Five mutated TRPC5 plasmids(<partinfo>BBa_K1943022</partinfo>, <partinfo>BBa_K1943023</partinfo>, <partinfo>BBa_K1943024</partinfo>, <partinfo>BBa_K1943025</partinfo>, <partinfo>BBa_K1943026</partinfo>).
Transient receptor potential channel 5 (TRPC5) is a mechanosensitive (MS) channel[1] which takes charge of hearing in our project. Future teams could use original TRPC5 plasmid (<partinfo>BBa_K1943021</partinfo>) to sense sound or other kinds of mechanical forces. Furthermore, we randomly mutated the ankyrin region of TRPC5 channel by PCR. We expected the ankyrin region to be a crucial part for sense mechanical forces based on the structure of TRPC5[2].(Detailed methods are shown in our notebook.)
These mutated TRPC5 plasmids(<partinfo>BBa_K1943022</partinfo>, <partinfo>BBa_K1943023</partinfo>, <partinfo>BBa_K1943024</partinfo>, <partinfo>BBa_K1943025</partinfo>, <partinfo>BBa_K1943026</partinfo>) have all been sequenced. Because of limited time, we have not yet transfected them into cells to observe whether the channels are more sensitive to sound and mechanical force or not. Future teams are welcomed to use and finish testing them.
The mutated amino acid sites are as listed:
| Part | Original amino acids | Amino acid after mutation |
|---|---|---|
| <partinfo>BBa_K1943022</partinfo> | Trs(240)+Ala(27) | Glu(240)+Thr(27) |
| <partinfo>BBa_K1943023</partinfo> | Ala(27) | Thr(27) |
| <partinfo>BBa_K1943024</partinfo> | Met(134)+Ala(27) | Ile(134)+Thr(27) |
| <partinfo>BBa_K1943025</partinfo> | Glu(236)+Ala(27) | Lys(236)+Thr(27) |
| <partinfo>BBa_K1943026</partinfo> | Gln(128)+Ala(27) | Arg(128)+Thr(27) |
TRPC5 sequence contains 1 EcoRⅠ RE site and 2 PstⅠ RE sites, which are unacceptable for BioBricks. Therefore, we did site-mutation by PCR for three times. Detailed information are shown in our notebook. Learn More
The primers we used are as listed:
| Mutated RE sites | Primer design |
|---|---|
| PstⅠ-1 | |
| PstⅠ-2 | |
| EcoRⅠ |
