Part:BBa_K3771050

J23100-sfGFP

Description

This composite part consists of the constitutive promoter J23100 (BBa_J23100) and sfGFP (BBa_K1321337). It serves as a positive control for another two oxidative-stress-induced promoters in our design, which should not alter its downstream gene expression under oxidative stress.

Usage and Biology

We compared the relative fluorescence intensity of sfGFP expressed by different promoters under the stimulation by different concentrations of paraquat. As expected, the sfGFP expression level of J23100 shows no difference between the groups with and without paraquat. P_soxS with its activator, SoxR (BBa_K37710XX), is highly activated under oxidative stress and has the highest gene expression level compared to J23100, P_katG (BBa_K37710XX), and P_soxS (BBa_K37710XX) solely (Figure 1).

Fig.1. Comparison of different promoter strengths by relative fluorescence intensity after 4.5-hour incubation with paraquat in various concentrations.

References

1. ItalianiVCS, Silva NetoJF, BrazVS, Marques MV. Regulation of Catalase-Peroxidase KatG Is OxyR Dependent and Fur Independent in Caulobacter crescentus. Journal of Bacteriology. 2011;193(7):1734-1744. doi:10.1128/jb.01339-10

2. Keseler IM, Mackie A, Santos-Zavaleta A, et al. The EcoCyc database: reflecting new knowledge about Escherichia coli K-12. Nucleic Acids Res. 2017;45(D1):D543-D550. doi:10.1093/nar/gkw1003
Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 73