Team:TJU China/Safety
SAFTY
Project Design
1. smURFP is nontoxic. SmURFP is a manual mutation which was developed from an allophycocyanin α-subunit (APCα). It is from Trichodesmium erythraeum. So we have good reasons to believe there isn't any safety problem about SmURFP.
2. BV is nontoxic. It is an intermediate in the metabolic process in hemoglobin and other heme containing species in animals. And it is also an important biochemical substance.
3. Three bacteria are belong to biosafety level 2. We chose Bifidobacterium, bacteroides fragilis and enterohemorrhagic E.coli as our expressing strain. There may be a little threat to human health, but the threat can be avoided in BSL-2.
4. Two constructions have no hazards. Both surface display and co-expression just enable bacteria to be detectable since it can emit fluorescence.
5. We completed the animal imaging experiment with the help of Nankai University, which has the ability to do animal experiments.
Lab Work
In order to avoid danger and train our team member, everyone accepted the biological safety test even some of our members in charge of wiki. Of course, they all qualified. Furthermore, we all also accepted the safety training on BSL-2, because some bacterial strains have a little threat to human health, and the threat can be avoided in BSL-2. What’s more, when we formally conducted experiments, every member will be supervised by our instructors and each other. If we found who conducted work nonstandardly, we will punish him/her by being on duty at that day. And all our waste are discarded safely, strictly followed stipulations of our school.
We first chose BL-21 (DE3) as our expressing strain, and we first constructed expression vectors in E. coli DH5α. In the process, use kits which are safe to extract, construct and transform our plasmids.
The organisms we use for later expression are safety level 1 and 2 organisms. Among the ones belonging to biosafety level 2, EHEC and Citrobacter rodentium, which are facultative anaerobes, are conducted in biosafety cabinet. And all the wastes are thrown separately after sterilization. Bacaeroides fragilis, which is also belongs to biosafety level 2, is obligate anaerobe. We conducted this strain in the anaerobic operating station, and all the waste are throw separately after sterilization, like EHEC and C. rodentium. Samples of these bacteria are also stored in a specific area away from crowded places.
We conducted our other operations on open clean benches and all the waste contains microorganisms are sterilized before further dispose to ensure safety. According to the situation, we will choose to wear protective equipment.
The following is our regulations of lab:
Be sure that you have worn protective equipment if necessary.
Be sure that you have informed the instructor of the next step of the experiment plan and gotten the permission of him.
Keep the super clean bench clean and operate correctly.
Properly handle the waste generated in the experiment.
Clean the table after the experiment.
Make the experiment record and finish experiment results in time.
Be sure that you have informed the instructor of the next step of the experiment plan and gotten the permission of him.
Keep the super clean bench clean and operate correctly.
Properly handle the waste generated in the experiment.
Clean the table after the experiment.
Make the experiment record and finish experiment results in time.
Shipment
To achieve safe shipment, we followed the iGEM official advice for Teams in China to send the samples to GenScript in Nanjin, China which is quick and convenient.
