Difference between revisions of "Part:BBa K899011"

Revision as of 23:08, 26 September 2012

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. The characterization can be found on our wiki page if you follow this link: http://2012.igem.org/Team:SDU-Denmark/labwork/Testing. 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 wasn't present (this enzyme produces the base that FFT polymerizes).

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.

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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