We aim to genetically modify unicellular microalga Chlamydomonas reinhardtii by intro=
ducing
nfsI gene of bacterium Enterobacter cloacae in order to
investigate how nitroreductase expressing-microalgae respond to trinitrotol=
uene
(TNT) exposure. Our experimental design is as follows: obtain a synthetic g=
ene
of nfsI with flanking prefix and
suffix of standard Biobricks, and ligate this insert to pRbcBRL, a vector w=
ith
appropriate expression and selection system for Chlamydomonas reinhardtii and obtain pRbcnfsI. Then, we can
transform Chlamydomonas reinhardtii=
with designed plasmid. After engineering microalgae, we will grow them in t=
he
presence of TNT and investigate effectiveness of nitroreductase activity on
biological degradation of TNT.
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SAFETY AND SECURITY QUESTIONS
1. Would the materials used in your project and/or your final product pose:
a. Risks to the safety and health of team members or others in the lab? Using hazardous materials always has risk to cause trouble for team and lab members. We always use hoods and wearing lab coat, gloves, goggles are essential for an experiment.
We are working on two species one of them is Chlamydomonas reinhardtii which has no dangerous effect on human health. Other one is Bacillus subtilis which also is not human pathogen but it could cause food poisoning.
b. Risks to the safety and health of the general public if released by design or accident? Chlamydomonas reinhardtii is a strain of algae and it could cause algal bloom that means it overgrown in a water ecosystem and become poisonous for other species which share same places.
According to a Toxic Substances Control Act report from the Environmental Protection Agency, Bacillus subtilis “is considered a benign organism as it does not possess traits that cause disease. It is not considered pathogenic or toxigenic to humans, animals, or plants. The potential risk associated with the use of this bacterium in fermentation facilities is low.” Its degree of toxicity is III and IV the lowest toxicity effect on other species.
c. Risks to environmental quality if released by design or accident? We haven’t checked modified organism is more competitive than natural strains. However; Bacillus subtilis is well-known and commonly used model organism like Chlamydomonas reinhardtii.
d. Risks to security through malicious misuse by individuals, groups or states? Our modified Chlamydomonas reinhardtii just degrade TNT and it could not use for terrorism activity. Those two modified organism could not have ability to damage human health. Our facility always has an active security system and it requires an identity card for enterance.
Please explain your responses (whether yes or no) to these questions.
Specifically, are any parts or devices in your project associated with (or known to cause): - pathogenicity, infectivity, or toxicity?
No - threats to environmental quality?
No - security concerns?
No
2. If your response to any of the questions above is yes:
a. Explain how you addressed these issues in project design and while conducting laboratory work. Natural strain cause algal bloom and threat environmental quality so we used biological hazardous waste for them. And after our work is finished, we applies bleach (%5) and then it goes to waste. b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry. Our parts have not any safety, security and health issues. However, adding NfsI gene into algae could make it more competitive than normal strains. It requires checking before releasing to nature.
3. Under what biosafety provisions will / do you operate?
a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible. Our institution have its own biosafety rules and it takes 19 page so we added a link below which leads to institution’s safety committee page and it contains two link; one of them orientation of our lab and other one is chemical disposal form.
http://unam.bilkent.edu.tr/UNAM%20Laboratory%20Safety.html
b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review. We have a Laboratory Safety Committee; however, we did not discuss about our project because our experiment contains standard cloning procedure, which has always done in the lab.
c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training. Our institution have an exam about biosafety, we passed it. It consists of orientation which mentioned above.
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