Regulatory
pCaiF

Part:BBa_K398326:Design

Designed by: Hugo Federico Cueto Rojas   Group: iGEM10_TU_Delft   (2010-06-14)
Revision as of 14:09, 27 October 2010 by Hugo 87 (Talk | contribs) (→‎Design Notes)

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Promoter of the CaiF protein


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
    COMPATIBLE WITH RFC[1000]


Design Notes

pCaiF is a natural promoter found in E. coli K12, pCaiF is part of the translational unit of the protein CaiF which is a transcriptional regulator of carnitine metabolism under anaerobiosis and glucose limitation. From the original sequence, we took the cAMP-Crp and RNApol sigma 70 binding sites. Under low glucose levels, the cellular cAMP (cyclic Adenosine Mononucleotide Phosphate) levels are high and thus Crp (Catabolite gene activator protein) will bind easier to this molecule forming the complex cAMP-Crp. This complex acts as a transcriptional regulator of over 180 proteins involved in metabolism of secondary carbon sources.

The original purpose of pCaiF for our project was to serve as a promotor for our proteins when glucose or other carbon sources easier to degrade are not in the medium, in that way cells will grow faster and once a certain cell density is reached they can start to degrade other carbon sources like alkanes.


Schematic drawing of the CaiF regulon in E. coli K12 (taken from [http://BioCyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU141 EcoCyc]).

This promoter binds to RNA polymerase, sigma 70 (sigma D) factor.

Schematic drawing of the RNApoly binding region of pCaiF.


And the sequence to which crp binds is the following:

Schematic drawing of the crp binding region of pCaiF.

Source

You can find the original sequence and more data about pCaiF in the following [http://BioCyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU141 link]

References

  1. EcoCyc [http://BioCyc.org/ECOLI/NEW-IMAGE?type=ENZYME&object=CPLX0-226]
  2. Eichler K, Buchet A, Lemke R, Kleber HP, Mandrand-Berthelot MA (1996). "Identification and characterization of the caiF gene encoding a potential transcriptional activator of carnitine metabolism in Escherichia coli." J Bacteriol 1996;178(5);1248-57. PMID: 8631699