Composite

Part:BBa_K4115039

Designed by: Kaijun Wang   Group: iGEM22_ShanghaiTech_China   (2022-09-30)
Revision as of 09:24, 3 October 2022 by Yurj (Talk | contribs)


J23101-B0034-luxR-B0015-Lux pR-B0034-sfGFP-B0015

J23101-B0034-luxR-B0015-Lux pR-B0034-sfGFP-B0015
Function gene regulator
Use in E.coli
RFC standard RFC 10, RFC 10 compatible
Backbone pBBR1
Submitted by (ShanghaiTech_China)

This composite part is used to detect the response of LuxR (BBa_C0062) to AHL signaling molecules. When LuxR receives the signal of AHL, it can regulate the downstream Lux pR (BBa_R0062) to make superfolder GFP (BBa_K4115000) express.

Then we can transform the plasmid into the A.caulinodans ORS571 to construct communications and regulations between E.coil and S.elongatus through quorum sensing signals.

Proof of function

We encapsulated E. coli with the CP and E. coli with LuxI into alginate gel beads. The E. coli with LuxI secreted the 3OC6 HSL signal molecule, and the E. coli with LuxR received the Signal and regulate the expression of downstream sfGFP, making it express. So we can detect green fluorescence in the latter E. coli under the irradiation of ultraviolet light.

LuxR regulations between E.coli.jpg

Figure 1: sfGFP is expressed under the regulation of LuxR between E.coli in a single gel bead. Two parallel groups are given and the above two are control groups.

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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 985


[edit]
Categories
Parameters
None