Difference between revisions of "Part:BBa K3739030"
Line 18: Line 18:
  
 
'''Fig.1.''' The result of colony PCR. Plasmid pET28a(+).
 
'''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.
 
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.

Revision as of 20:02, 21 October 2021


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.

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.

T--XMU-China--K3739030 (K3739030 pET28a, regular PCR).png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 406
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 258
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 157
    Illegal SapI site found at 475
    Illegal SapI.rc site found at 838