Difference between revisions of "Part:BBa K1122680"

 
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<partinfo>BBa_K1122680 short</partinfo>
 
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This part enables expression of ferric uptake repressor (Fur) using IPTG induction
 
This part enables expression of ferric uptake repressor (Fur) using IPTG induction
  
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===Usage and Biology===
 
===Usage and Biology===
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1122680 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1122680 SequenceAndFeatures</partinfo>
  
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===Cloning===
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Part was cloned from ''B. subtilis'' gDNA. Reaction product was analysed on an agarose gel (Figure 1)
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Figure 1. Agarose gel shows a PCR product of a desired size
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===Functional analysis===
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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
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[[File:Fur_graph]]
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Figure 2. Survival of cells induce and noninduced to express Fur in varying iron concentrations.
  
 
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Revision as of 16:26, 4 October 2013

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)

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

File:Fur graph

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