Device

Part:BBa_K1475010

Designed by: Daniel Weltz Pedersen   Group: iGEM14_SDU-Denmark   (2014-10-07)
Revision as of 20:17, 15 October 2014 by Danie12 (Talk | contribs)

dxs+LIMS1+appY regulated by the tet promoter

This part contains the dxs gene from Escherichia coli encoding the thiamine-diphosphate dependant 1-deoxyxylulose-5-phosphate synthase, the LIMS1 gene from Citrus limon encoding (R)-limonene synthase ([http://www.brenda-enzymes.org/enzyme.php?ecno=4.2.3.20 BRENDA:EC 4.2.3.20]) and the appY ([http://www.uniprot.org/uniprot/P05052 UniProt:P05052]) gene from Escherichia coli encoding a transcriptional regulator related to anaerobic metabolism. These genes are under the control of the TetR promoter.
Overexpression of 1-deoxyxylulose-5-phosphate synthase ([http://www.brenda-enzymes.org/enzyme.php?ecno=2.2.1.7 BRENDA:EC 2.2.1.7]) and appY has been shown to increase production of carotenoids and other terpenoids [1] (limonene is a terpenoid) and has thus been included in this part by the 2008 Edinburgh team, find the original part here: Part:BBa_K118024.


Usage and Biology

The dxs gene from Escherichia coli encodes the 1-deoxyxylulose-5-phosphate synthase, which requires thiamine-diphosphate to function. It catalyses the conversion of pyruvate and D-glyceraldehyde 3-phosphate into 1-deoxy-D-xylulose 5-phosphate and CO2. [1][2]

The LIMS1 gene encodes the (R)-limonene synthase from Citrus limon. (R)-limonene synthase conversion of geranyl diphosphate into (R)-limonene and diphosphate. [3]

appY is a transcriptional regulator that induces the transcription of acid phosphatase (appA) together with other proteins. appY transcriptional repression functions. [4]


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 SapI.rc site found at 808
    Illegal SapI.rc site found at 2068


References

1. Leighanne A. Brammer et al.: 1-Deoxy-d-xylulose 5-Phosphate Synthase Catalyzes a Novel Random Sequential Mechanism. J Biol Chem. Oct 21, 2011; 286(42): p. 36522–36531. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3196101/
2. BRENDA, 2014: EC 2.2.1.7 - 1-deoxy-D-xylulose-5-phosphate synthase. http://www.brenda-enzymes.org/enzyme.php?ecno=2.2.1.7&Suchword=&organism%5B%5D=Escherichia+coli&show_tm=0
3. BRENDA, 2014: EC 4.2.3.20 - (R)-limonene synthase. http://www.brenda-enzymes.org/enzyme.php?ecno=4.2.3.20&Suchword=&organism%5B%5D=Citrus+limon&show_tm=0
4. UniProt, P05052- APPY_ECOLI, http://www.uniprot.org/uniprot/P05052

[edit]
Categories
Parameters
None