Coding

Part:BBa_K3170602

Designed by: Haodong Yan   Group: iGEM19_LZU-CHINA   (2019-10-20)


OR 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 removed the target sequence of tobacco etch virus protease (TEVp), the target sequence of tobacco vein vein virus protease (TVMVp) and YFP on the right side of the CFP with the terminator removed, and combined them into OR Boolean logic gates.
  • In the OR logic gate, when both TEVp and TVMVp proteases are absent, FRET occurs, and the fluorescence emitted by the CFP absorption excitation light is absorbed by YFP, and the YFP emits yellow 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 CFP is separated from the YFP. The two are far apart, failing to perform FRET, and YFP does not emit light. The CFP absorbs the excitation light and emits cyan fluorescence.
  • For OR 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

T--LZU-CHINA--OR.png
  • OR 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 or logic gate pathway shows that when TEVP and TVMVP are present alone, the fluorescence value at 515 nm is high, and when both are present, the fluorescence value is very low, indicating that two proteins are required to function in the pathway alone.

Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 694
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 740
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 764
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None