Difference between revisions of "Part:BBa K398331"

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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.
 
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.
  
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[[Image:T--SEU-Nanjing-China--GROUP.png|600px|center|<b>Figure 1.</b> ABTS assay cuvettes after 16 hours at room temperature, containing 100µl culture supernatant (clones 1-14, day 2), 100µl 2mM ABTS and 800µl citric acid-phosphate buffer pH 4.0.  Cuvette labelled 31 contains only ABTS and buffer, cuvette labelled 32 contains only supernatant and buffer. Cuvette labelled C contains the wild-type control supernatant.]]
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[[File:T--SEU-Nanjing-China--GROUP.png|200px|thumb|left|alt text]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 01:58, 6 October 2019

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.

alt text

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