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Regulatory

Part:BBa_K950000

Designed by: Jakob Matthes   Group: iGEM12_Tuebingen   (2012-07-10)

yeast fet3 promotor

The FET3 protein is an essential element of Saccharomyces cerevisiae’s iron metabolism. FET3 is a multicopper oxidase that is responsible for high-affinity ferrous iron (Fe(II)) uptake in yeast (Askwith et al., 1994). The promoter of the FET3 gene (Pfet3) of Saccharomyces cerevisiae is metalloregulated, i.e. repressed at high intracellular iron levels (Lyons et al., 2004).

Interestingly, Pfet3 is also repressed by active membrane progesterone receptors (Smith et al., 2008) - see Parts BBa_K950006 and BBa_K950007 - and other proteins of the PAQR family (Kupchak et al, 2007).

 

Basal expression level of mOrange under the control of the promotors Pfet3, Panb1 and Psuc2. Non-transformed yeast was used as negative control. Fluorescence was detected using a RFP filter set (ET Bandpass 470/40, ET Bandpass 572/35).

 

Alterations of mOrange expression in low-iron environment of yeast strain W303 containing Pfet3-mOrange-pTUM100. Fluorescence was measured using a plate reader (Ex: 548nm +/-9nm, Em: 581nm +/-20nm) and normalized to OD600 of the cell suspension. All measurements were performed as duplicates. As shown in the diagram, the promotor activity of Pfet3 has increased after 4 h of incubation in low-iron medium compared to cells which were grown in normal synthetic complete medium. After 8 h of incubation the activity had increased 3-fold.


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
    COMPATIBLE WITH RFC[1000]



[edit]
Categories
//chassis/eukaryote/yeast
//promoter
//regulation/negative
Parameters
None