Difference between revisions of "Part:BBa K4115039"

Revision as of 12:43, 4 October 2022

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

This composite part is used to detect the response of LuxR (BBa_C0062) to AHL signaling molecules [1]. When LuxR receives the signal of AHL, it can regulate the downstream Lux pR (BBa_R0062) to make superfolder GFP (BBa_K4115000) express.

This part is togther used with LuxI (BBa_C0161), an AHL(3OC6 HSL here) sender.We can transform the plasmid into the A.caulinodans ORS571 to construct communications and regulations system between E. coli and S.elongatus HL7942 through quorum sensing signals.

Proof of function

We encapsulated E. coli with this composite part 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.

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.

Quantitative analysis

We set three groups,one is the blank group, where only E. coli with LuxR is added, the second is the group where 10^-7M 3OC6 HSL is added, and the third one is the group where the supernatant of LuxI is added. We conclude the data by dividing the fluorescence intensity by the OD600 and get Figure 2.

Figure 2: Different 3O6C HSL concentrations are inputed to test LuxR response intensity.

Sequence and Features

References

[1] Kylilis, N., Tuza, Z.A., Stan, GB. et al. Tools for engineering coordinated system behaviour in synthetic microbial consortia. Nat Commun 9, 2677 (2018). https://doi.org/10.1038/s41467-018-05046-2