Part:BBa_I718002:Experience

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Applications of BBa_I718002

• We transformed DGAT gene in pBluescript SK minus vector (Stratagene) (ampicilline resistance and pLac promoter) (pKS::DGAT) into E.coli chemically competent (DH5alpha). We used Nile Red fluorescence dye in the medium (5µg/mL medium) to observe lipid inclusions in different conditions (0.4mM IPTG induction in LB medium with or without sodium oleate 2mM).
  

- Line 1 represents E.coli transformed with pKS::DGAT; and Line 2 the negative control (E.coli transformed by part B0015).
- Columns 1 and 2 are LB medium without sodium oleate supplementation; columns 3 and 4 are LB with sodium oleate supplementation (2mM).
- Columns 1 and 3 are without IPTG; columns 3 and 4 with IPTG induction (0.4mM).

We can observe lipid inclusion into E.coli transformed by pKS::DGAT with IPTG induction. With or without adding oleate we do not observe significant differences.

• We could think that only dead cells are fluorescent (DGAT could kill the cells) but with cell death marker (green) we can see that cell death is not increased with DGAT.

User Reviews

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