Composite

Part:BBa_K568001

Designed by: Team TU Munich 2011   Group: iGEM11_TU_Munich   (2011-09-01)
Revision as of 18:55, 21 September 2011 by Thorsten mueller (Talk | contribs)

Optogenetical AND-Gate red/blue light

This logical gate is based on amber stop-codon suppression via the non-canonical tRNA supD (BBa_K228001). The blue light sensor (BBa_K238013) induces the expression of a mRNA coding for a T7-polymerase with an amber mutation (BBa_K228000), that can only be translated by ribosomes if the correct supD tRNA is present. The supD tRNA is expressed when the red light sensor (BBa_K568000) is induced. Thus, only cells which receive both signals produce the desired substances.

design

Usage and Biology

If the part is hit by both far red light (705 nm) and blue light (465 nm) beams, the AND-Gate is turned on. Red light induces the autophosphorylation at the cytosolic site of cph8. This leads to phosphorylation of OmpR which subsequently binds to OmpC promotor and enables transcription of the supD t-RNA. Blue light leads to dimerisation of YcgF and binding of the repressor YcgE. The formation of the YcgE-YcgF complex leads to the unbinding of YcgE from the YcgZ promoter which activates the transcription of T7ptag (T7 polymerase with the amber stop codon mutation) if enough supD tRNA is available.

This AND-gate should ensure that expression of T7ptag is only induced when both wavelengths hit bacteria with the part.

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
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 644
    Illegal XhoI site found at 1786
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 971
    Illegal AgeI site found at 3817
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3820

Functional Parameters

n/aOptogenetical AND-Gate red/blue light

Induction with red light at 705 nm

Induction with blue light at 465 nm

Shut down of constant red light signalling with 650 nm

[edit]
Categories
Parameters
n/aOptogenetical AND-Gate red/blue light