Difference between revisions of "Part:BBa K1149009"

 
 
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<partinfo>BBa_K1149009 short</partinfo>
 
<partinfo>BBa_K1149009 short</partinfo>
  
Degrades PLA.
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Encodes a lipase that hydrolyses compound polylactic acid.
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Source organism: Cryptococcus sp. strain S-2
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<h2>Characterisation</h2>
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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.
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https://static.igem.org/mediawiki/2013/thumb/5/59/CLE.png/450px-CLE.png
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'''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.
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https://static.igem.org/mediawiki/2013/thumb/d/d8/CLE_fluor.png/450px-CLE_fluor.png
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'''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.
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<b>Conclusion: There is no growth inhibition caused by induction, nor is there any significant fluorescence.</b>
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<h2>References: </h2>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

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.

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 in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

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 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]