Team:METU Turkey/safety.html

1.Would any of your project ideas raise safety issues in terms of:

researcher safety,

There are no hazardous chemicals that we used in the laboratory except for EtBr that we used in a special room for gel visualization and some buffers used in commercially available kits.

public safety, or

Since all the organisms used in the wetlab are destroyed after they are done working with they pose no danger for public safety. We use E.coli strain DH5alpha which is easy to kill and all the student members who work in the wetlab are very well trained to make sure there is no unwanted contamination. This year we are also working on a kill switch which, if works properly, will eliminate any bacteria whereas they will not be able to survive without IPTG present.

environmental safety?

As mentioned above a kill switch will be available in the organism which will initiate cell death when the cell is in an environment that does not contain enough IPTG.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

We have followed all the safety procedures during the isolation process from the cells we had and we successfully isolated the DNA without any problems that will raise safety issues.

3.Is there a local biosafety group, committee, or review board at your institution?

If yes, what does your local biosafety group think about your project?

If no, which specific biosafety rules or guidelines do you have to consider in your country?

In our university we have a biosafety and ethical research center which monitors the safety of our projects. http://www.ueam.metu.edu.tr The members of METU Biology department are also members of National Biosafety Coordinating Committee. We are able to consult to them whenever it is necessary. Also biosafety is a very important subject for METU Department of Biology, whereas the student members are very well trained in the subject. Also the projects that we decided to work on are presented to many people in the area in order to see if they are of any concern for the biosafety regulations. All the procedures that are used in the lab are also approved procedures that are used by these people mentioned above.

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

We believe using specific strains designed specifically for labwork will be safer. It is possible to genetically modify the organisms so that, they are not able to grow outside the lab environment. There probably is not a way to build some biobricks that can be modified in order to make them safer since their function will be the cause of safety issues.

EVEN MORE QUESTIONS! (Thanks to iGEM ATOMS Turkiye)

Questions?

1) When you change the genetic information of bacteria in bees’ guts this may have the possibility of changing the genetic material of most bacterial flora. Have you thought about a possible solution to this?

2) Will this change effect the bees honey production and so affect humans and animals health?

3) How will you manage the amount of Coumaric Acid produced?

4) Will Coumaric Acid affect the plants which the bees touch?

Answers!

1) We will introduce our gene construct to the bacteria before hand and then introduce it to the host bee via food (a process called paratransgenesis), so it is highly unlikely for the other bacteria to get this part. Also, our system is optimized for B. subtilis, the promoters we used and the products as a whole have slim to no chance of functioning anywhere else.

2) The bacillus strain we selected is native to the bees' gut. It is normally not found in the honey stomach of the bee, even if the bacteria were to get into honey, it will still be in the natural levels. Other than that, according to literature, IPTG is never traced in honey, so our little germs, even if they can make it into honey, will be killed because of our kill switch.

3) There is actually no real data about the p-coumaric acid levels let alone it is not known if such a thing as p-coumaric acid overdose exists. We used in our system a constitutive promoter for the production of it too. What we expect and what we could argue from literature is that having p-coumaric acid is in all cases preferable. Maybe exceeding levels could cause increased energy consumption and maybe even acid accumulation, a decrease in tyrosine(a precursor of p-coumaric acid) etc. What most probably will happen (and what we actually hope for) is that the natural feedback mechanism will be triggered and control the process. But you are right, a inductive system would have been prettier

4) P-coumaric acid is mostly produced by plants. Even vanilla production and cinnamon flavour begins with p-coumaric acid. Many plants produce p-coumaric acid so p-coumaric acid exposure should not cause any harm to them. Besides, p-coumaric acid will be produced in the midgut of the bee, so it is unlikely for p-coumaric acid to find its way directly to the plant.