Composite

Part:BBa_K2505030:Design

Designed by: Hazuki Hasegawa   Group: iGEM17_TokyoTech   (2017-10-17)


Ptet-rbs-traI (K34G)-tt


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The gene traI(K34G) is derived from Agrobacterium tumefaciens and encode a enzyme necessary for synthesizing Quorum Sensing signaling molecules ([N-]acyl-homoserine lactones, AHLs), 3OC8 HSL (hereafter C8), in E. coli. This part constitutively produces C8. We introduced point mutation to traI gene and the productivity of C8 was improved by approximately 3-fold. We introduced this part to E. coli then E. coli could produced enough C8 to induce transcription of human cells.

The mutation was introduced wild type traI(BBa_K553001).

The DNA sequences of traI (K34G) is optimized for expressing in E. coli considering the codon usage.


Source

Mutation was newly introduced on the BBa_K553001 by PCR.

References

[1] Pavan Kumar Reddy Kambam, Daniel J. Sayut, Yan Niu, Dawn T. Eriksen, Lianhong Sun (2008) Directed evolution of LuxI for enhanced OHHL production. Biotechnology and Bioengineering Volume 101, Issue 2 1 October 2008 Pages 263-272

[2] MATTHEW R. PARSEK, DALE L. VAL, BRIAN L. HANZELKA, JOHN E. CRONAN, E. P. GREENBERG (1999) Acyl homoserine-lactone quorum-sensing signal generation. Proc. Natl. Acad. Sci. USA Vol. 96, pp. 4360-4365, April 1999 Biochemistry