Composite

Part:BBa_K899011

Designed by: Andreas Kjær   Group: iGEM12_SDU-Denmark   (2012-09-26)
Revision as of 23:03, 26 September 2012 by Andkj11 (Talk | contribs)

FFT (BBa_K899001) with double RBS (BBa_K899009)

This is a composite part containing our fructan:fructan fructosyltransferase (FFT) gene with a "double" RBS containing both the shine-dalgarno and kozak sequence. This is the main part of our project, and also what we submitted to iGEM. The part was characterized in a pJET vector (http://www.bioinfo.pte.hu/f2/pict_f2/pJETmap.pdf) and we found a weird looking substance that resembled sugar structure in the growth medium. When we expressed FFT in E. coli top 10 strains they grew as fast as regular top 10 bacteria, probably because the sucrose:sucrose fructosyltransferase (SST) enzyme wasnt present (this enzyme produces the base that FFT polymerizes).

SDUiGEM-2012-Graf_GC.png

When staining for inulin we observed a slight increase in inulin even though the SST enzyme wasn't present. This is probably because there was something in the substrate that enabled FFT to polymerize the sugar.

<center>InulinStainingpic2.png

Figure 2 - From top to bottom, left to right: Top10 negative test, top 10 with inulin added, FFT, SST. First 2 rows is the first round, third and fourth row indicates second experiment. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 534
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 388
  • 1000
    COMPATIBLE WITH RFC[1000]


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