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  To protect the bees against living microorganisms that are dangerous for the bees is our aim. We made a research and we realized that bees recently suffer from insufficient feed.

Since beekeepers prefer to feed the bees with high-fructose corn syrup or sucrose because of the low prices, bees started to suffer from insufficient nutrition. Bees lost their ability of fighting against the pathogens, parasites and pesticides and this situation creates a major problem, colony losses. P-coumaric acid is the most important substance as one of these nutritions.

P-coumaric acid, a monomer of sporopollenin, is the essential substance for the bees’ diet.

 In this year, in the kill switch part of our project, we use an mRNA interferase, MazF, which cleaves mRNA’s at specific sequences. In our kill switch, we used anti-sense RNA principle as a template. According to this principle, MazF, which is constantly produced via a constitutive promoter, is got inactivated by Anti-MazF construct. In order to trigger this mechanism, we used IPTG, a harmless molecule for the bee which at the same time does not appear in the honey too. When IPTG is present in the environment, LacI is inhibited by IPTG and therefore promoter gets activated. With the activated promoter of it, Anti-MazF is produced and inactive MazF. As long as IPTG exists, MazF gets inactivated continuously; therefore, the bacteria maintain their lives. On the other hand, if bacteria exists in a IPTG-free environment, Anti-MazF producing stops, which leads to MazF producing and bacteria get killed by MazF.

In this circuit, we used the parts K143053 as a constitutive promoter and SpoVG RBS due to their more efficient works in Bacillus subtilis. Besides these two, we used the part I732820 to produce LacI, but we had to change the RBS of this system since RBSelowitz is less effective in Bacillus then RBSspoVG. With this change in the activity of the RBS, we can compete with the production of MazF.