Part:BBa_K3490011
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE 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 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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