Part:BBa_K4781000:Design
A DSF (Diffusible Signal Factor) sensing module
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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
Experimental verification
After assembly, SDS Page was performed to check the presence of these proteins
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.