Composite
GFP-XylE

Part:BBa_K316006:Experience

Designed by: IC 2010 Team   Group: iGEM10_Imperial_College_London   (2010-10-21)
Revision as of 00:16, 28 October 2010 by Chris (Talk | contribs)

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K316006

  • This part can be used as an inducible reporter enzyme.GFP was added to the N-terminus of C2,3O monomer. The C2,3O enzyme is an obligate homotetramer where all 4 subunits are required for its activity. By adding GFP at the N-terminus of the each monomer, tetramerization of the enzyme is blocked, thus rendering it inactive. This modification is important as it makes the system inducible. The cleavage of the GFP by TEV protease is the input inducer signal, means that Catechol dioxygenase monomers are released so that they can form the active tetramer.

User Reviews

UNIQe90f93f1c0fe6432-partinfo-00000000-QINU

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Imperial College iGEM 2010

  • There is about a 10-fold reduction in the rate at which the protein product of GFP-XylE gene construct catalyses the reaction in comparison to the wild type XylE gene.
Graph shows production of HMS (yellow product) over time after catechol addition at time 0 minutes. Different curves represent different catechol concentration added to the cell cultures
Graph shows production of HMS over time after catechol addition at time 0 minutes. Different colored curves represent different substrate concentrations added at time 0min in cell cultures.
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