Signalling

Part:BBa_K2033002:Design

Designed by: Brady Dennison   Group: iGEM16_Arizona_State   (2016-08-15)
Revision as of 23:51, 13 October 2016 by Jixu2471 (Talk | contribs)


isovaleryl-HSL, 3-methyl-N-[(3S)-2-oxooxolan-3-yl]butanamide Sender- BjaI


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 294
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The BjaI part arises from the soil bacterium Bradyrhizobium japonicum. The designed part by Ryan Muller was cloned into competent DH5AT E. coli cells. These were ligated into the psb1C3 vector and plated, producing the following gel:

T--Arizona State--Gel4.jpg

An optical density test was conducted on the produced BjaI construct to determine if the AHL is produced. The plate reader ran an 8-hour read from 580-610nm, indicating the presence of the mCherry fluorescent molecule. The AHL gene lies upstream of the mCherry gene, so successful production of mCherry is a good indicator that the AHL molecule is being produced. A positive growth curve was found for the BjaI construct over the 8-hour read.

T--Arizona State--BJARFP.png

Source

Bradyrhizobium japonicum

References

Lindemann, A., G. Pessi, A. L. Schaefer, M. E. Mattmann, Q. H. Christensen, A. Kessler, H. Hennecke, H. E. Blackwell, E. P. Greenberg, and C. S. Harwood. "Isovaleryl-homoserine Lactone, an Unusual Branched-chain Quorum-sensing Signal from the Soybean Symbiont Bradyrhizobium Japonicum." Proceedings of the National Academy of Sciences 108.40 (2011): 16765-6770. Web.