Team:OUC-China/Safety

1. Would any of your project ideas raise safety issues in terms of: researcher safety, public safety, or environmental safety?

About the safety risks on team members or others in the lab，though risks to the safety and health of team members do exist, we think this can be avoided and reduced to the least by careful and standard operation. When doing molecular experiments, researchers can hardly avoid hazardous reagents (like EB and other toxic organic solvent) or physical radiation. Without careful and proper training, experimenters could be harmed. But by strictly following the laboratory safety manual and the lab regulation of Tsingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Science, we will ensure safety of ourselves and other people sharing the lab with us.

On preliminary consideration, our project has no threat to the public and environment but may cause small-scale ecological disorder. This worry will be described in detail in the question two.

We do not think our project or any part of the design have great potential to be misused maliciously by individuals, group or state. Because at present time, this work is just conveying a philosophical theory and remains to be developed to make contributions in other fields. But we can not ensure that someone intelligent enough will not drive the work onto an evil way. In fact, every project can be misused when encountered with such a person or group. Should it happened, it is not the inventor’s fault.

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

In our project, the system uses five kinds of Lux homologous QS gene, three of which are from the Rhizobium sp., which can invade their host plant and form nodulation, some of these QS genes may have relationships with Ti plasmid transferring, but luckily, real transferring only occurs when these genes are combine with other essential genes like traABC, which are not included in our system, hence it will be almost impossible for the E.coli to contaminate the other organisms’ genomes. Another potential worry is the cinI, one of the AHL synthase, which can produce 3OH-C14:1-HSL. This molecule is a kind of “small bacteriocin” which is able to inhibit the growth of other rhizobia, it may cause small-scale ecological disorder among rhizobia and its host.

Above we expressed our worries about the bad effect of our project. However, we are mostly positive about our project’s biosafety on researchers, public and environment. In fact, Theory of five elements itself, where the idea of our project comes from, is to explain the importance of balance, harmony and good will.

Firstly, our final product will work in nutritionally deficient E.coli strain—HB101, which can not survive once released.

Then, before we transfer the final design into the nutritionally deficient strain HB101, we have used several other strains for plasmid cloning, expressing and going test. DH5α, a strain who is thi- and F- ,cannot produce in a wild environment without thiamine (this situation is common) and could not transfer its genes into other organisms. DH10βand BL21 are also F-. The possibility of genes being released into the environment is small.

Last, in our laboratory operation, we are very careful when dealing with these gene-modified E.coli . Usually, we will wear white lab coats and latex gloves always. The growing E.coli plate will be sealed with elastic adhesive tape when in incubator or refrigerator. The deserted E.coli medium will be washed with the 84 disinfectant. The deserted plate will be sealed in package and put into a big beaker (in case the melting solid medium stains the autoclave sterilizer) to be sterilized. All of these can prevent the modified E.coli from being released and causing unexpected result.

We still have some concerns about the use of antibiotic. Recently, many pan-drug resistant bacteria have appeared because of the unrestricted use of antibiotics and have posed great threats to public health. As we know, for convenience, we usuallwill use antibiotic in research process. most of the construction of two biobricks will not avoid include antibiotic use; we wonder whether some careless overuse of it may raise unexpected dangerous mutation in the bacteria. However, we haven’t found a more definite way to avoid it.

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

Both Ocean University of China and the CAS obey the “General Requirements on Biosafety in Laboratory” (GB19489-2004) released by our country People’s Republic of China and follow the “Laboratory Biosafety Manual” issued by WHO in 2004, the third edition. Besides we have our own specific biosafety rules. Lab security regulation rules of Experimental Teaching center of Marine Life Sciences of OUC can be find [http://222.195.158.132/syzx/013126.htm here]. while the specific lab rule of QIBEBT is not available online but is carried out in laboratory all the time. We do not have a institutional biosafety committee but own a equivalent group, they are composed of our three professors in OUC and two researchers in QIBEBT, plus several doctors and masters in OUC and QIBEBT have given us some advice. They guide us not only in experiment but also in biosafety questions. After reviewing, they think our project are basically qualified in terms of researcher health, public and environment safe. We didn’t receive adequate biosafety training before, only knowing its importance from our professional class. But every one of us have received some lab trainings more or less. Four junior students and two sophomores of us have received both microbiology lab training and molecular biology lab training. The rest have received the microbiology lab training, either. Besides, we participated in a lot of other biological lab training when in amateur time, like ecology, taxonomy, biochemistry, immunology and so on. Our country People’s Republic of China had issued a document “General Requirements on Biosafety in Laboratory” (GB19489-2004) in April the 5, 2004, it can be viewed [http://wenku.baidu.com/view/cd5837d9ad51f01dc281f13a.html here].

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 recommend IGEM headquarter hold a small laboratory biosafety training before all iGEMers starting their summer work. If possible, we recommend all iGEM teams contribute to draft the competition’s own “iGEM-laboratory biosafety manual” and then all the iGEMers will follow this common standard. Hope IGEM competition got better day by day.

As for parts, devices and system’s biosafety, we recommend all the iGEMers use safe chassis for transformation and expression. For parts associated with pathogenicity, infectivity, or toxicity, we should better add “LVA” tags to them, or at least design devices limiting them in the same chassis.