Part:BBa_K3170600

NOR Boolean logic gate

Compotent of the protease-protease regulation system

• One of the logic gates of the core protein pathway designed using Boolean logic. Some of them separated Porcine Parvovirus (PPVp) and then reconstituted by Link short sequence; the other part separated YFP from the terminator and linked to PPVp target sequence and CFP. The two parts are then joined by P2A, a self-cleaving small peptide, and combined into a NOR Boolean logic gate.
• In the NOR logic gate, when both TEVp and TVMVp proteases are absent, the C-terminus and N-terminus of PPVp are combined into PPVp, the PPVp target sequence is cleaved, and CFP and YFP are separated, which are far apart and cannot be FRET. YFP does not emit light, and the CFP absorbs the excitation light and emits cyan fluorescence. When any one of the proteases or both proteases is present, the target sequence of one of the proteases or the two proteases is cleaved, and the C-terminal and N-terminal portions of the PPVp cannot be combined into PPVp. Therefore, the PPVp target sequence is not cleaved, FRET occurs, the fluorescence emitted by CFP absorbs the excitation light is absorbed by YFP, and YFP emits yellow fluorescence.
• For NOR logic gates, there are two inputs: the presence or absence of two proteases, TEVp and TVMVp, when the protease is present, ie the input value is 1, otherwise 0; there is an output: CFP luminescence or not, when CFP illuminates, The output is 1, otherwise it is 0.

Experimental Results

• NOR logic gate. The reporter was used at a concentration of 150 ng, and the fluorescence value was measured at 515 nm. The norm gate showed that the ordinate was the normalized fluorescence intensity value. When TEVP and TVMVP are present, the fluorescence value at 515 nm is very low, and when both are absent, the fluorescence value is high, indicating that it is necessary to have two proteins at the same time in the pathway to function.

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 2996
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 283
  Illegal BamHI site found at 1066
  Illegal BamHI site found at 2264
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2288
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1102
  Illegal BsaI.rc site found at 1282