Part:BBa_K3127996

This part contains LasI ，LasR ，CcaR，CcaS，ho1，pcyA and ccdB.

It is a part of the Light Controlled Bacteria Co-culture system.This part contains LasI ，LasR ，CcaR，CcaS，ho1，pcyA and ccdB. When receiving the green light, this part can release 30C6HSL and produce CcdB protein.

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 658
  Illegal NheI site found at 715
  Illegal NheI site found at 1577
  Illegal NheI site found at 1600
  Illegal NheI site found at 3388
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2007
  Illegal XhoI site found at 1431
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3089
  Illegal NgoMIV site found at 3749
  Illegal NgoMIV site found at 5490
  Illegal AgeI site found at 3946
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 6358
  

Characterization

Figure 1/2 shows the trends of each bacteria in the LCBC-system. We got the data through the enzyme-labelled instrument and analyzed it to get the figure. The y-axis is the number of cells, and the x-axis is fluorescence intensity. There are two kinds of E.coli. One kind of E.coli carried sfGFP on the genome. The other kind of E.coli carried RFP on the genome. Cells were individually cultured in the LB medium for 12 hours before common inoculated in 12-well plates in LB medium in the first 13 hours. Figure 1 shows the different fluorescent intensity of E.coli under the light of 640-650 nm wavelength for the first 13 hours and the light of 460-465 nm wavelength for the next 12 hours.Figure2 shows the different fluorescent intensity of E.coli under the light of 640-650 nm wavelength for the first 13 hours and the light of 530-535 nm wavelength for the next 12 hours.

Figure 6: We can see that after 12 hours of blue light exposure, the number of E. coli bacteria carrying RFP dropped by half.

Figure 7: After 12 hours of green light exposure, the number of E.coli carrying GFP decreased by half.

Figure 3/4 shows the change in the ratio of two bacteria in LCBC system. We used CLSM to observe and process data. The second row shows the ratio of the two bacteria which have been exposed to the light of 640-650 nm wavelength for 12 hours and then to the light of 460-465 nm wavelength for the same time. The third row shows the proportion of the two bacteria which have been under the light of 640-650 nm wavelength for 12 hours and the light of 530-535 nm wavelength for another 12 hours.