We were inspired to create this project by the Tsinghua iGEM 2017 team. This team created a two part recognition system to recognize aflatoxin B1 in plants, using coordinately bound single chain variable fragment antibodies fused to either the activating domain or binding domain of Gal4 to mediate activation of pGal1 mediated reporter genes. We saw the development of this technology as an opportunity to create recognition systems for a number of toxins in plants, and therefore wanted to validate a similar experiment in plants. We have been working this year to increase the reporters and the ease with which our expression system can be modified using a series of interchangeable parts, and have just begun experiments to validate our plasmids/reporter constructs. To create the interchangeable parts, we created a gateway compatible multiple cloning site, into which we could clone in and out promoters and reporters in the PGWB1 and PGWB2 systems. We are testing our reporters without the use of toxin in Nicotiana benthamiana using agrobacterium mediated transient expression. We have completed making our system accessible to interchangeable promoters and transgenes, but still need to prove the efficacy of the reporters/promoters.
It is our hope that future plant biologists will be able to use the vector systems, reporters, and recognition devices we create.