Part:BBa_K4645016

J23100-B0030-vqmA-B0017-Pqtip-tetR-B0015-PTet-amcyan-B0017

Verification of Not-Gate.

Usage and Biology

We transformed this biobrick into E. coli to regulate expression of genes. With the growing of bacteria, DPO accumulates. While the concentration of DPO reached the threshold value, the expression of NeuA, NeuB and TetR would be activated. Then TetR shut down the expression of amcyan in downstream.

Figure 1. The circuit we assembled.

Characterization

Blank BL21, BL21 transformed with this biobrick, were cultured in microplate reader 37°C, 220 rpm for 10 hours and detected OD600, fluorescence intensity (458 nm excitation light, 489 nm emission light) every 10 minutes.

Figure 2. Fluorescence intensity / OD600 in BL21 and luxR-tetR

The result showed that in the early growth stage, transformed bacteria expressed amcyan fluorescent protein continually. When the group density reached the threshold value, the expression of amcyan fluorescent protein began to be blocked up.

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
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 599
  Illegal AgeI site found at 2547
  
• 1000
  COMPATIBLE WITH RFC[1000]