Difference between revisions of "Part:BBa K1149009"

Latest revision as of 23:42, 2 October 2013

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 in LB 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 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.

References:

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 99
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 464
Illegal AgeI site found at 1229
1000
COMPATIBLE WITH RFC[1000]

@@ Line 11: / Line 11: @@
 https://static.igem.org/mediawiki/2013/thumb/5/59/CLE.png/450px-CLE.png
-'''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 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 in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
 https://static.igem.org/mediawiki/2013/thumb/d/d8/CLE_fluor.png/450px-CLE_fluor.png
-'''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.
+'''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 in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
 <b>Conclusion: There is no growth inhibition caused by induction, nor is there any significant fluorescence.</b>