Composite

Part:BBa_K4732001

Designed by: Matthew Hanson   Group: iGEM23_Alma   (2023-10-10)
Revision as of 14:13, 12 October 2023 by Matt 2001 (Talk | contribs)


DDT biosensor (RtER-araC-RFP)

RFP under the control of Rainbow trout estrogen receptor by way of araC: This is our biosensor for xenoestrogens like DDT. In the presence of such molecules, rtER will prevent expression of the araC gene, which in turn relieves repression of RFP. In this case, araC is an inverter module allowing for a red color signal in response to the xenoestrogen.

Characterization of DDT biosensor - Alma 2023

The DDT biosensor works by rtER binding to DDT (which has characteristics similar to estrogen). This causes for the estrogen response element to bond to the receptor stoping transcription of the araC repressor. The araC repressor stops expression of RFP. With no transcription of the araC repressor, RFP can be transcribed and expressed. Tests to determine if the parts of the DDT biosensor worked. Arabinose is a sugar that represses the araC repressor. Due to this, red fluorescence should be seen from the biosensor. Initial tests used 5% arabinose at different volumes added to six different plates, then left over night. The results of the plating are shown in the figure below:


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1914
    Illegal NheI site found at 1937
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 908
    Illegal BamHI site found at 570
    Illegal BamHI site found at 3161
    Illegal XhoI site found at 464
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 3774
    Illegal AgeI site found at 3886
  • 1000
    COMPATIBLE WITH RFC[1000]


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