Coding

Part:BBa_K200006:Design

Designed by: Royah Vaezi, David Roche   Group: iGEM09_Imperial College London   (2009-08-12)
Revision as of 14:09, 1 September 2009 by Droche (Talk | contribs)

OtsB: Part 2 of 2 coding for trehalose producing enzymes

Sequence codes for trehalose phosphatase enzyme. This enzyme is the second of two required for the conversion of glucose to trehalose.
This enzyme catalyses the following reaction:
alpha,alpha-trehalose 6-phosphate + H2O -> alpha,alpha-trehalose + phosphate

Trehalose is a disaccharide formed from two glucose molecules. Throughout nature, trehalose is associated with resistance to dessication and cold shock [http://www.pnas.org/content/99/15/9727.full.pdf+html 1], and is naturally produced in Escherichia Coli. We hope that by upregulating the trehalose production pathways in E.coli we can increase trehalose concentrations within our cell, thereby conferring some resistance to protein degredation in our system. This would allow easy transport and storage of the final product.



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 455
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PCR from Escherichia Coli BL21(DE3)

Primers Used:
Forward:
GCTCTAGATGACAGAACCGTTAACCG
Recomended Temperatures for PCR : 54.5C(without overhang) and 64.8C(with overhang)

Reverse:
GGACTAGTATTAGATACTACGACTAAACGAC
Recomended Temperatures for PCR : 54.7C(without overhang) and 64.2C(with overhang)

Biobrick Overhang shown in Bold

Source

Coding sequence isolated from e-coli

References