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Our Project Description:   Taking a major role in pollination, bees are one of the most important organisms within an ecosystem. However their populations are in serious decline. Colony Collapse Disorder has been found as the most common cause of the disappearance of bees in large numbers. Some pathogens like parasitic fungi and a pesticide called imidacloprid are the leading reasons of CCD. In this study, our aim is to decrease the number of hives affected by chemical compounds such as imidacloprid. Our plan is to turn the mutualistic bacteria living in bees' guts into a shield mechanism to protect the bees against these factors. A protein CYP6G1 found in Drosophilia melanogaster has the ability to degrade imidacloprid into harmless substances. Moreover, coumaric acid increases the general immunity of bees against harmful components and parasitic organisms and we aim to increase the level of coumaric acid in bees' guts. The main objective of this study is the transformation of the genes coding for these two proteins to Bacillus subtilis, which mutualistically live in bees' guts.

As a well-known fact, bees take the major part in pollination, which leads to give fruit, while preparing honey. During the last few decades, some powerful insecticides like imidacloprid were developed to fight with harmful insects; however, the ability to kill almost all insects started to affect other insects, which they must not have affected from the beginning.

This was the start point of our project. We, as a group formed by mostly biological sciences students, knew that no ecological system can maintain without bees. When we looked into the source of the problem, it was clearly revealed that we couldn’t stop usage of imidacloprid. Then, we came up with an idea to protect bees against existing imidacloprid. Having known that drosophila has some sort of immunity to this insecticide, we planned to use it.

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.

Imidacloprid Immunity

Immune System Boost

Kill Switch

Tweets by @iGEM_METU2013

First meeting of METU iGEM '13 team

It was a snowy day when we fell in love with iGEM.

The aseptic conditions are satisfied.

Researching for our project.

Optimizations for lab work

Collaboration with ITU iGEM team @ Istanbul

Seminar about synthetic biology given by us to METU College students

Hanging out with Baskent Uni iGEM team

Modelling team is working, not playing LoL.

Participating in Treasure Hunt '13 in METU

Modelling team saying goodbye to our captain, Buse Reis, who kept in touch with us in Cambridge.

Strengthening our bonds by group activities

The collaboration of all iGEM teams in Turkey. We presented our projects to each other and our advisors to improve them and to get feedback.

Making an introduction to lab works for ITU MOBGAM team

#direnplate

One of our meetings with Assoc. Prof. Dr. Mesut MUYAN

Collaboration and education with mechanical engineers of METU

Ancient autoclave found in METU Biological Sciences building

Planning the circuits

Hosting future iGEMers from Güven Eğitim & Sağlık Vakfı in METU

Young iGEMers appreciating science

Our captain after doing hard laboratory work :D

SCIENCE B*TCH

Enjoying our bee cookies aseptically in lab

Thanks to Nihal'in Elinden for cookies

Enjoying our bee cookies not so very aseptically in lab

Falcon quality control :p

Modelling team at work

Raising awareness about synthetic biology in METU

Brainstorming for the project

SynBio Party poster

SynBio Party cocktails

SynBio Party people having fun

SynBio Party

SynBio Party 300 -500

SynBio Party

SynBio Party :D

SynBio Party which made us some funds

Synthetic Biology Day Crew

Our beloved participants in Synthetic Biology day

Synthetic Biology Day Crew and Participants

iGEM Stand in METU

iGEM stand in METU

Keep Calm and Follow Us

Hülooooğğğğ

Safety first :D

Click here to follow us at Facebook!

Click here for our national site!

Click here for our blog!

Click here for even more photos!

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-	<p id="u3292-2"><span id="u3292">{font-size: 65%}&nbsp;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&#45;sense RNA principle as a template. According to this principle, MazF, which is constantly produced via a constitutive promoter, is got inactivated by Anti&#45;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&#45;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&#45;free environment, Anti&#45;MazF producing stops, which leads to MazF producing and bacteria get killed by MazF.</span></p>
+	<p id="u3292-2"><span id="u3292">&nbsp;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&#45;sense RNA principle as a template. According to this principle, MazF, which is constantly produced via a constitutive promoter, is got inactivated by Anti&#45;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&#45;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&#45;free environment, Anti&#45;MazF producing stops, which leads to MazF producing and bacteria get killed by MazF.</span></p>
-	<p id="u3292-4">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.</p>
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