Difference between revisions of "Part:BBa K3490011"

 
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For biosafety, we use TetR-VP16(Tet-off) system to repress the expression of lysis. The tet-off system makes use of TetR protein to associate with VP16 protein first then bind with tetracycline and further bind to TetO region to repress the expression of Lysis. Once the bacteria leak out the contact lenses which will lack of tetracycline further induce bacteria to express lysis. Hence, we can prevent the bacteria from causing damage to our eyes.
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Our first kill switch design used TetR fused with VP16, tet-off system<sup>[1]</sup> to control the Colicin E7(BBa_K117000) as our biosafety measure. We will put tetracycline analog into the contact lens, this will prevent the VP16 to activate the lysis gene. If the bacteria leave the contact lens, where the environment has low concentration of tetracycline analog, the lysis gene will be activated.(Fig.1)
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<br>There are several options when it comes to tetracycline analog that can act on the tetR protein, changing its DNA binding affinity, however, according to Biomol GmbH-Life Science Shop, 50 mg of Anhydrotetracycline costs 54 US dollars[2]. In search of a cheaper alternative, we turned to tetracycline(TC)[3] and Chlortetracycline(CTC)[4], which has been reported to bind to <i>tetR</i> protein and affect its affinity to <i>tetO</i>.
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<img src="https://static.igem.org/mediawiki/parts/a/ad/T--NCKU_Tainan--BBa_K3490011.png" style="width:35%;">
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<br>Fig. 1 Our design about the killswitch using tet system
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:30, 25 October 2020


Causing bacteria lysis when the bacteria leak out the contact lenses.

Our first kill switch design used TetR fused with VP16, tet-off system[1] to control the Colicin E7(BBa_K117000) as our biosafety measure. We will put tetracycline analog into the contact lens, this will prevent the VP16 to activate the lysis gene. If the bacteria leave the contact lens, where the environment has low concentration of tetracycline analog, the lysis gene will be activated.(Fig.1)
There are several options when it comes to tetracycline analog that can act on the tetR protein, changing its DNA binding affinity, however, according to Biomol GmbH-Life Science Shop, 50 mg of Anhydrotetracycline costs 54 US dollars[2]. In search of a cheaper alternative, we turned to tetracycline(TC)[3] and Chlortetracycline(CTC)[4], which has been reported to bind to tetR protein and affect its affinity to tetO.



Fig. 1 Our design about the killswitch using tet system





Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 755
    Illegal NheI site found at 1068
    Illegal NheI site found at 1091
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]