Revision as of 19:37, 27 September 2013 by Fanxiao0606 (Talk | contribs)

PPD Switching System

We integrated the PPD sensor and the PPD reporter system by Tet-off system. AHL induction activates the expression of tTA in PPD sensor system, after which tTA trans-activates the downstream PPD reporter ADE2, realizing the connection between input AHL signal and output ADE2 signal.

  • Overview
  • Design
  • Discussion


We connected the PPD sensor and reporter system by Tet-off system.

We transformed the newly constructed PPD sensor and the PPD reporter in to a and α haploid yeasts respectively.

We realized trans-activating the reporter part by yeast mating.

Figure 1. Overview of PPD switching system


In order to link the sensor and reporter system by Tet-off system. We replace the mCherry in pTF4 by TetR-VP16. The expression of TetR-VP16 was under the control of cyc100 mini promoter. We constructed pTF5 plasmid.

Figure 2. pTF5

After demonstration of feasibility of pTF6, we created a reporter plasmid in pRS415. pRS415 is a single copy plasmid with centrosome used in leucine auxotrophic yeast. The plasmid fragment from TRE to CYC1 terminator was inserted in to pRS415. We got pTF7 plasmid.

Figure 3. pTF7



[1] F.Sherman et al. Getting Started with Yeast By Fred Sherman, 2003, MethodsEnzymol. 350, 3-41.