Difference between revisions of "Part:BBa K1149006"
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| + | <h2>Characterisation</h2> | ||
| + | Our PulA construct was tested for growth upon induction by addition of Arabinose. | ||
| − | + | https://static.igem.org/mediawiki/2013/thumb/c/c1/PueB.png/450px-PueB.png | |
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| + | '''PulA growth assay.''' ''E. coli'' (MG1655) transformed with PulA (BBa_K1149006) were grown with either 0 μM or 6 μM Arabinose to induce PulA expression. There was no growth inhibition as p = 0.7648 > 0.05, therefore induction did not cause significant growth decrease. Growth was at 37°C 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 in PulA. </b> | ||
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| + | <h2>References: </h2> | ||
<p>[http://www.sciencedirect.com/science/article/pii/S0964830598000687 Cloning and expression in Escherichia coli of a polyurethane-degrading enzyme from Pseudomonas fluorescens]</p> | <p>[http://www.sciencedirect.com/science/article/pii/S0964830598000687 Cloning and expression in Escherichia coli of a polyurethane-degrading enzyme from Pseudomonas fluorescens]</p> | ||
Revision as of 22:39, 2 October 2013
pelB-pulA-flag expression
Expression of a plastic degradation enzyme from Pseudomonas fluorescens under arabinose inducible promoter. The protein in codon optimized for expression in ecoli, is flag tagged and has an Nterminal pelB tag for secretion. Encodes polyurethane esterase, break down ester bonds in polyurethane.
PUR degradation by esterase:
Characterisation
Our PulA construct was tested for growth upon induction by addition of Arabinose.
PulA growth assay. E. coli (MG1655) transformed with PulA (BBa_K1149006) were grown with either 0 μM or 6 μM Arabinose to induce PulA expression. There was no growth inhibition as p = 0.7648 > 0.05, therefore induction did not cause significant growth decrease. 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 in PulA.
References:
[http://www.sciencedirect.com/science/article/pii/S0964830598000687 Cloning and expression in Escherichia coli of a polyurethane-degrading enzyme from Pseudomonas fluorescens]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 804
Illegal BamHI site found at 65 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1099
- 1000COMPATIBLE WITH RFC[1000]