Team:Wageningen UR/Basic Part

Wageningen UR iGEM 2016



Basic parts

Our best basic part: BBa_K1913025

BBa_K1913025 encodes the plac-FixK2 hybrid promoter. The sequence of upper control element and core element region are derived from the wildtype FixK2 promoter from the bacterium Bradyrhizobium japonicum.
BBa_K1913025 can be be activated by phosphorylated FixJ, part of the YF1-FixJ light sensor. In the presence of light, FixJ is no longer phosphorylated and BBa_K1913025 is not activated. At the same time, it can be inhibited by the LacI repressor because of two lac operators that are positioned on both sides of the FixK2 promoter core.
The BBa_K1913025 was tested together with the YF1-FixJ light sensor using mRFP as a reporter. After culturing for 24 hours in darkness, BBa_K1913025 has five times higher fluorescence than a control without the light sensor.
In our project, the plac-FixK2 promoter is part of the toggle switch. This switch is intended for regulating Bt toxin expression. The promoter is only activated in darkness, inside the beehive. Furthermore, it stays inactivated through LacI mediated repression until B12 is sensed, which inhibits the formation of LacI (see also our project about detecting Varroa mites)

Below, you can find all basic parts we created!

BioBrick number BioBrick name Designer
BBa_K1913000 chiA for Varroa destructor Lisa
BBa_K1913001 chiB for Varroa destructor Lisa
BBa_K1913008 vitamin b12 riboswitch Carina
BBa_K1913009 Guanine riboswitch Carina
BBa_K1913019 Guanine riboswitch BS-yxjA Tianhe
BBa_K1913020 mRFP with degredation tag Tianhe
BBa_K1913021 sGFP with defredation tag Tianhe
BBa_K1913022 Synthetic plac-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913023 Synthetic plac-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913024 Synthetic ptet-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913025 Wild type plac-FixK2 hybrid promoter Tianhe
BBa_K1913026 Wild type ptet-FixK2 hybrid promoter Tianhe