Measurement

Part:BBa_K398331

Designed by: Kira Schipper   Group: iGEM10_TU_Delft   (2010-10-07)
Revision as of 01:51, 6 October 2019 by Bread (Talk | contribs) (Team SEU-Nanjing-China's 2019 Characterization)

pCaiF-B0032 measurement device

pCaiF is a promoter region from E.coli containing the binding site for the global regulator Crp which is expressed during periods of starvation. Under, for example, glucose limiting circumstances pCaiF activates transcription. pCaiF is tightly regulated by cAMP levels.


The characterization and results of BBa_K398331 are described on the BBa_K398326 (promoter) BioBrick page and on the [http://2010.igem.org/Team:TU_Delft/Project/sensing TU Delft iGEM Team 2010 wiki]



Team SEU-Nanjing-China's 2019 Characterization

[http://https://2019.igem.org/Team:SEU-Nanjing-China Team SEU-Nanjing-China ] measured this part BBa_K398331.Our aim is to characterize the pCaiF promoter. We are very interested in this promoter. According to the description of this part, pCaiF is a natural promoter found in E. coli K12 , which regulates the expression of genes involved in the degradation of non-glucose carbon sources. The promoter is regulated by the level of cAMP. Because decomposition product of glucose auses decrease of cAMP, on the contrary, at low glucose level, the level of cAMP increases. CRP binds to cAMP to form complex cAMP-CRP. cAMP-CRP binds to pCaiF and activates the transcription of downstream components. Therefore, we hope to observe the experimental results: the expression of GFP in high glucose should be lower than that in low glucose.

1、We prepared LB medium with 5 different concentration of glucose, each concentration containing triplicate. The 5 concentration of glucose are 0.5g/L, 1g/L, 2g/L, 5g/L, 10g/L, LB medium without glucose as control. The E.coli grew in 15mL medium.

T--SEU-Nanjing-China--GROUP.png
T--SEU-Nanjing-China--GROUP.png


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.rc site found at 722


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