Team:BGU Israel/Design
Our main goal in the project was to find a better delivery system for biological pest control, since today's treatments mainly focus on large water sources, without regard to smaller water sources, such as plant pots. In order to do so, we asked ourselves the question: Who knows best where mosquitoes lay eggs? The obvious answer was the female mosquito herself. We thought, why not use the female mosquito as our guided missile, and she will direct us to the smaller water sources. She will be our "Trojan Mosquito."
We created an initial masterplan that goes as follows:
1. Create a polycistronic plasmid that will contain the toxic subunits of Bacillus thuringiensis subsp. israelensis (Bti).
2. Using electroporation, insert the toxic polycistronic plasmid into Serratia marcescens bacteria in order to create transgenic bacteria.
3. Feed the Serratia bacteria to female mosquitoes.
4. Let female mosquitoes mate with their male companions.
5. The female mosquitoes will lay infected eggs.
6. These infected eggs will hatch into larvae.
7. The hatched larvae will die from the expressed toxin and will infect its immediate surrounding water source.
After consulting with various experts, our plan changed from infecting female mosquitoes, to infecting male mosquitoes (To know more, you can check out our "Integrated Human Practice").
And so, we changed a key aspect of our plan:
1. Create a polycistronic plasmid that will contain the toxic subunits of Bacillus thuringiensis subsp. israelensis (Bti).
2. Using electroporation, insert the toxic polycistronic plasmid into Serratia marcescens bacteria in order to create transgenic bacteria.
3. Feed the Serratia bacteria to male mosquitoes.
4. Let male mosquitoes mate with their female companions, infecting the females.
5. The female mosquitoes will lay infected eggs.
6. These infected eggs will hatch into larvae.
7. The hatched larvae will die from the toxic polycistronic plasmid and will infect the surrounding water source.
In order to achieve our plan, we divided our experimental design to subcategories:
1. Delivery of Bacteria- Inspect if Serratia indeed integrates into the mosquito's microbiome and pass on to the eggs.
2. Plasmids Constructions- The plan and construct of our polycistronic plasmid.
3. Toxicity Assays- Check if our initial plasmids containing the Bti subunits indeed kill mosquito larvae.
4. Bonuses- You will have to read further in order to find out!