Composite

Part:BBa_K1149006

Designed by: Margarita Kopniczky   Group: iGEM13_Imperial_College   (2013-09-13)
Revision as of 23:46, 2 October 2013 by Jps09 (Talk | contribs)

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: PUR_esterase_theoretical_pathway_2.jpg

Characterisation

Our PulA construct was tested for growth upon induction by addition of Arabinose.

450px-PueB.png

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 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 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 804
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1099
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//function/degradation
Parameters
None