Part:BBa_K4781000:Design

A DSF (Diffusible Signal Factor) sensing module

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 398
  Illegal NgoMIV site found at 2669
  Illegal AgeI site found at 1230
  Illegal AgeI site found at 1709
  Illegal AgeI site found at 1793
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 447
  Illegal BsaI.rc site found at 311
  

Design

Had to ensure that the sensory domain of RpfC of Xanthomonas Campestris is similar to the sensory domain of the RpfC of Xanthomonas oryzae. Had to conduct blast to examine its similarity. Similarly blast was conducted to compare RpfG too.

Usage

File:Aiim-dete.png

This two-component regulatory system was used to sense DSF and subsequently was modified to use it for the detection of DSF leading to its reporting with the purple color.

Experimental verification

After assembly, SDS Page was performed to check the presence of these proteins

File:SDS-PAGE-igem-iiser-kolkata.png

This two-component regulatory system was used to sense DSF and subsequently was modified to use it for the detection of DSF leading to its reporting with the purple color.

Source

Used an inducible promoter to express RpfC and RpfG protein under a strong RBS which will work together to form the two component regulatory system that senses DSF.

References

1. Slater, Holly, et al. "A two‐component system involving an HD‐GYP domain protein links cell–cell signalling to pathogenicity gene expression in Xanthomonas campestris." Molecular microbiology 38.5 (2000): 986-1003.