Part:BBa_K3170605

AND Boolean logic gate

Compotent of the protease-protease regulation system

• One of the logic gates of the core protein pathway designed using Boolean logic. We will remove the target sequence of tobacco etch virus protease (TEVp) and the target sequence of tobacco vein vein virus protease (TVMVp) from the CFP of the terminator, and then add a YFP to each of the TEVp target sequence and the TVMVp target sequence. (Where the left YFP does not contain a terminator), combined into an AND Boolean logic gate.
• In the AND logic gate, when both TEVp and TVMVp proteases are absent, FRET occurs, CFP absorbs the excitation light and the fluorescence emitted by YFP is absorbed, and YFP emits yellow fluorescence. When two proteases are present, the TEVp target sequence and the TVMVp target sequence are cleaved, CFP and two YFPs are separated, YFP is far from CFP, and FRET cannot be performed between the two, so YFP does not emit light, and CFP absorbs at this time. Excitation light emits cyan fluorescence. When only one protease is present, one of the protease target sequences is cleaved, one YFP is separated, and the other YFP is linked to CFP, FRET occurs, the fluorescence emitted by CFP is absorbed by YFP, and YFP emits yellow fluorescence.
• For AND 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 is illuminated The output value is 1, otherwise the output value is 0 if the CFP does not emit light or the light is weak.

Experimental Results

• AND logic gate. The reporter was used at a concentration of 150 ng, the fluorescence value was measured at 515 nm, and the ordinate was the normalized fluorescence intensity value. The and logic gate pathways show that when TEVP and TVMVP are present alone, the fluorescence value at 515 nm is very low, and when both are present, the fluorescence value is high, indicating that both proteins are required to function in the pathway.

Usage and Biology

Sequence and Features

Assembly Compatibility:

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

  Illegal NotI site found at 1449
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]