Composite

Part:BBa_K1897015:Design

Designed by: Ang Shi Hui   Group: iGEM16_NUS_Singapore   (2016-10-10)


LuxR + LuxI


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 1852
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 50
    Illegal AgeI site found at 1065
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There is one HA tag available for characterisation of the LuxR and LuxI proteins respectively produced via western blot. Note that the stop codons for LuxR BBa_C0062 and LuxI BBa_C0061 is shifted to after the HA tag.

Also, the transcriptional terminators rrnBT1 (from BBa_B0010) + BBa_B0012 for LuxR was derived from BBa_B0015, with the first 8 base pairs removed.

Source

LuxR was obtained from biobrick part BBa_C0062, which was derived from Vibrio fischeri. The promoter was synthesized based on the sequence obtained from BBa_J23119. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

LuxI was obtained from biobrick part BBa_C0061, which was derived from Vibrio fischeri. The promoter was synthesized based on the sequence obtained from BBa_R0062. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

References

Choi, S. H., & Greenberg, E. P. (1992). Genetic evidence for multimerization of LuxR, the transcriptional activator of Vibrio fischeri luminescence. Molecular marine biology and biotechnology, 1(6), 408-413.

Fuqua, W. C., Winans, S. C., & Greenberg, E. P. (1994). Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators. Journal of bacteriology, 176(2), 269.

Shadel, G. S., & Baldwin, T. O. (1991). The Vibrio fischeri LuxR protein is capable of bidirectional stimulation of transcription and both positive and negative regulation of the luxR gene. Journal of bacteriology, 173(2), 568-574.

Trott, A. E. (2000). Amino Acid Residues in LuxR Critical for its Mechanism of Transcriptional Activation during Quorum Sensing (Doctoral dissertation, Virginia Polytechnic Institute and State University).