Regulatory

Part:BBa_K950003

Designed by: Jan Rudolph   Group: iGEM12_Tuebingen   (2012-07-29)

yeast suc2 promotor

In Saccharomyces cerevisiae the expression of SUC2 genes (which encode invertase) is heavily repressed by high levels of glucose (Lutfiyya and Johnston, 1996). The proteins which repress SUC2 genes in this case are MIG1 and MIG2 whereby MIG1 is a stronger repressor than MIG2 (Lutfiyya and Johnston, 1996). MIG1 and MIG2 both are zinc-finger-containing repressor proteins with binding-sites in the SUC2 promoter (Psuc2). MIG1 has its highest affinity to the GC-rich SUC2-A site which is between position -505 and -483 in the SUC2 promoter (Nehlin and Ronne, 1990, Lutfiyya and Johnston, 1996). The release of repression is mediated by the protein kinase SNF1 (sucrose nonfermenting) in yeast (Celenza and Carlson, 1986, Carlson, 1998).

The MIG1 gene is also available from the registry! See Part BBa_K950009.

 

Increased promoter activity of Psuc2 and Panb1 in repressor deficient strains. Fluorescence was detected using a RFP filter set (ET Bandpass 470/40, ET Bandpass 572/35).

 

Promoter activity of Psuc2 and Panb1 in repressor deficient strains. The promoter activity was enhanced by a factor of 2.6 for Psuc2 and 13.8 for Panb1. 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 triplicates.

 

Alterations of mOrange expression upon galactose induction of yeast strain W303 containing Psuc2-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.

 

Alterations of mOrange expression upon galactose induction of mig1 deficient yeast containing Psuc2-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.


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