Part:BBa_K3739030
blrA-GFP
blrA can be activated by blue light and produce reactive oxygen species(ROS).
Biology
Flavin-combined fluorescent protein, coded by a kind of optogenetic toolbox LOV(light-oxygen voltage)-based photosensitizer gene, can induce the light-driven killing of bacteria. BlrA, originating from Bacillus subtilis, is a blue light-induced GTP-binding receptor, which possesses the LOV domain and autofluorescence. LOV domain enables it to be activated by blue light and produce ROS, which can damage the bacterial structure.
Gfp codes for a green fluorescence protein.The fluorescence density of GFP can reflect the expression level of BlrA.
Usage
To intuitionally show whether blrA expresses or not, it is linked to a GFP through a 3GS linker. The growth condition and the level of BlrA can be reflected by the intensity of green fluorescence.
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Fig.1. The result of colony PCR. Plasmid pET28a(+).
We take blrA as the core of our kill switch, combine it with promotor, terminator and a series of necessary elements. The whole part is inserted into a plasmid backbone and finally transformed into E.coli to gain enough correct and functional circuits. The circuits are transformed into Vibrio.natriegens. We choose growth curve and CFU to measure the killing effect of BlrA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 406
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 258
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 157
Illegal SapI site found at 475
Illegal SapI.rc site found at 838
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