Part:BBa_K1379004

S. pneumoniae σ^x CDS

Gene sequence of competence protein comX which produce SigmaX factor. This sigmaX is the inducer for PcelA promoter (BBa_K1379002), and Phelicase promoter (BBa_K1379003) encoding GFP. This sigmaX protein will bind to 8 base pairs of PcelA or Phelicase promoter and trigger gene expression. The sigmaX gene was cloned from E. coli NCTC 7465 strain.

Characterization

To test the functionality of Sigma X protein, Construct consisting SigmaX generator-PcelA promoter-GFP generator (BBa_K1379005), and SigmaX generator-Phelicase promoter-GFP generator (BBa_K1379007) are built. In order to prove that SigmaX induce these two promoters, GFP expression were observed under microscope with positive control (BBa_I20260), and negative control consisting only PcelA-GFP without SigmaX generator BBa_K1379002, and Phelicase-GFP without SigmaX generator BBa_K1379003

Figure 1. PcelA and Phelicase promoter induced by SigmaX protein drives GFP expression. While the same construct without SigmaX protein did not give any GFP signals. Another negative control which is only protein sigmaX without PcelA or Phelicase also did not give any GFP signals. Reference promoter BBa_J23100 + GFP is used as positive control. Scale bar = 5mm.

Sequence and Features

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 298