Difference between revisions of "Part:BBa K3771050"

 
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<br><b style="font-size:1.3rem">Description</b>
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<br>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.
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===Usage and Biology===
 
===Usage and Biology===
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<br><b style="font-size:1.3rem">Usage and Biology</b>
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<br>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. <i>P<sub>soxS</sub></i> with its activator, SoxR (BBa_K37710XX), is highly activated under oxidative stress and has the highest gene expression level compared to J23100,  <i>P<sub>katG</sub></i> (BBa_K37710XX), and <i>P<sub>soxS</sub></i> (BBa_K37710XX) solely (Figure 1).
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<img src="https://static.igem.org/mediawiki/parts/c/c2/T--NCKU_Tainan--sfGFP_Expression_%28PQ_%404.5h%29.png"​ style="width:35%;">
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<p align="center"> Fig.1. Comparison of different promoter strengths by relative fluorescence intensity after 4.5-hour incubation with paraquat in various concentrations.
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<br><b style="font-size:1.3rem">References</b>
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<br>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
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<br>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
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3771050 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3771050 SequenceAndFeatures</partinfo>

Revision as of 15:14, 17 October 2021


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. PsoxS with its activator, SoxR (BBa_K37710XX), is highly activated under oxidative stress and has the highest gene expression level compared to J23100, PkatG (BBa_K37710XX), and PsoxS (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