Part:BBa_K1149013
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.
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 250
Illegal AgeI site found at 1195 - 1000COMPATIBLE WITH RFC[1000]
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