Translational_Unit

Part:BBa_K1122680

Designed by: Aleksandra Lewicka, Jan Lyczakowski   Group: iGEM13_Edinburgh   (2013-08-30)

Plac - LacZ - Fur

This part enables expression of ferric uptake repressor (Fur) using IPTG induction


Usage and Biology

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 BsaI site found at 769

Cloning

Part was cloned from B. subtilis gDNA. Reaction product was analysed on an agarose gel (Figure 1)

Fur gel.png

Figure 1. Agarose gel shows a PCR product of a desired size

Functional analysis

It was assumed that over-expression of Fur could increase bacterial tolerance to increased iron concentration due to enhanced perception of environmental [iron]. This was analysed experimentally be culturing cells induced and not induced to express Fur in LB nmedium supplemented with increasing concentrations of iron. Results obtained (figure 2)indicate that indeed the cells that contain more copies of Fur protein survive better in medium with increased iron levels

Fur graph.png

Figure 2. Survival of cells induce and noninduced to express Fur in varying iron concentrations.


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Parameters
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