Difference between revisions of "Part:BBa K1149013"
 
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bdh2: converts 3HB to acetoacetate
 
bdh2: converts 3HB to acetoacetate
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<h2>Characterisation</h2>
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Our bdh2 (extracellular) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Arabinose.
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https://static.igem.org/mediawiki/2013/thumb/f/ff/Bdh2.png/450px-Bdh2.png
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'''Bdh2 with pelB secretion tag growth assay.''' ''E. coli'' (MG1655) transformed with pelB-bdh2 BBa_K1149013 were grown with either 0 μM or 6 μM Arabinose to induce bdh2 and sfGFP expression. Bdh2 induction shows reduced growth of MG1655 but after 6h they reach the same OD, the decrease in growth is confirmed as not significant by a two-tailed t-test where p = 0.6964 > 0.05 (critical value). Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
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https://static.igem.org/mediawiki/2013/thumb/a/ab/Bdh2_fluor.png/450px-Bdh2_fluor.png
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'''Bdh2 with pelB secretion tag induction assay.''' ''E. coli'' (MG1655) transformed with pelB-bdh2 BBa_K1149013 were grown with either 0 μM or 6 μM Arabinose to induce bdh2 and sfGFP expression. We see that induction leads to stronger expression of bdh2 than without, although the promoter is leaky as the curve follows a similar pathway. Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
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<b>Conclusion: There is no growth inhibition caused by induction, nor is there any significant fluorescence induction.</b>
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<h2>References: </h2>
  
 
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Latest revision as of 23:47, 2 October 2013

bdh2: converts 3HB to acetoacetate

bdh2: converts 3HB to acetoacetate

Characterisation

Our bdh2 (extracellular) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Arabinose.

450px-Bdh2.png

Bdh2 with pelB secretion tag growth assay. E. coli (MG1655) transformed with pelB-bdh2 BBa_K1149013 were grown with either 0 μM or 6 μM Arabinose to induce bdh2 and sfGFP expression. Bdh2 induction shows reduced growth of MG1655 but after 6h they reach the same OD, the decrease in growth is confirmed as not significant by a two-tailed t-test where p = 0.6964 > 0.05 (critical value). Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

450px-Bdh2_fluor.png

Bdh2 with pelB secretion tag induction assay. E. coli (MG1655) transformed with pelB-bdh2 BBa_K1149013 were grown with either 0 μM or 6 μM Arabinose to induce bdh2 and sfGFP expression. We see that induction leads to stronger expression of bdh2 than without, although the promoter is leaky as the curve follows a similar pathway. Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

Conclusion: There is no growth inhibition caused by induction, nor is there any significant fluorescence induction.

References:

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 250
    Illegal AgeI site found at 1195
  • 1000
    COMPATIBLE WITH RFC[1000]