Part:BBa_K1149009
Cutinase-Like Enzyme
Encodes a lipase that hydrolyses compound polylactic acid.
Source organism: Cryptococcus sp. strain S-2
Characterisation
Our Cutinase-like enzyme (CLE) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Xylose.
CLE growth assay. E. coli (MG1655) transformed with CLE BBa_K1149009 were grown with either 0% or 2% Xylose to induce CLE and sfGFP expression. Induction with xylose shows growth inhibition. Ideally you would normalise the fluorescence with OD600 to give fluorescence/cell in order to get the actual difference in induction expression. A two-tailed t-test shows that p = 0.4308 > 0.05, therefore there is no statistical difference between induced and uninduced. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
CLE induction assay. E. coli (MG1655) transformed with CLE BBa_K1149009 were grown with either 0% or 2% Xylose to induce CLE and sfGFP expression. CLE induction does not seem to increase the amount of fluorescence emitted by the GFP. Growth was at 37°C 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.
References:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 99
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 464
Illegal AgeI site found at 1229 - 1000COMPATIBLE WITH RFC[1000]
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