Part:BBa_K2012002

PleD from Caulobacter crescentus, a response regulator with a diguanylate cyclase (DGC) domain.

Intracellular c-di-GMP (Hengge 2009) concentration has been regulated by two functionally opposing enzymes, the diguanylate cyclases (DGCs) containing a GGDEF domain, and phosphodiesterases (PDEs) containing either an EAL or HD-GYP domain.

PleD (Wassmann, Chan et al. 2007) from Caulobacter crescentus, a response regulator with a diguanylate cyclase (DGC) domain, in its activated form.

Figure 1. Expression of pleD. For demonstrating expression of pleD, we used Congo red staining assay. As previous mentioned, high concentration of c-di-GMP could induce E. coli synthesize exopolysaccharides, and Congo red binding is a complex phenotype that reflects various outer membrane and surface properties including the presence of adhesive structures such as curli fimbria which are involved in biofilm formation. Wild type: DE3 contain pET28b plasmid, colony which was stained red color contain pET-pleD plasmid.

Reference:

Hengge, R. (2009). "Principles of c-di-GMP signalling in bacteria." Nat Rev Microbiol 7(4): 263-273.

Wassmann, P., et al. (2007). "Structure of BeF3- -modified response regulator PleD: implications for diguanylate cyclase activation, catalysis, and feedback inhibition." Structure 15(8): 915-927.

Assembly Compatibility:

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

  Illegal BamHI site found at 14
  Illegal BamHI site found at 1172
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 430
  Illegal NgoMIV site found at 577
  Illegal AgeI site found at 913
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 87