Difference between revisions of "Part:BBa K2938018"

Revision as of 13:22, 12 October 2019

Polycistronic BTI Toxin

PR1 - RBS - Cry4Ba (HA tag) - RBS - Cry11Aa (Strep tag) - RBS - Cyt1Aa (Strep tag) - RBS - P20 (His tag) - RBS - deGFP - T500 Terminator.

We planned this plasmid to be the most toxic to the Aedes aegypti mosquito larvae. Containing the most toxic Bacillus thuringiensis subsp. israelensis (Bti) subunits.

In order to build this plasmid, we constructed three initial plasmids [BBa_K2938014, BBa_K2938015, BBa_K2938016] which can build this final plasmid using gibson assembly. Our 3 initial plasmids showed toxicity against the mosquito larva (which you can see under experiences page of the plasmids), and the final plasmid should show better toxicity due to synergistic activity of the different subunits.

During the gibson assembly of the initial plasmids we added tags to each of the subunit in order to see it's expression using western blot, and we placed deGFP downstream of the polycistronic plasmids, closest to the terminator in order to follow our GFP marker in confocal microscope meaning the upstream toxic genes were also expressed.

Sequence and Features