Team:NCKU Tainan/Safety
Safety
Safe Laboratory
Before stepping into the laboratory, all the members in our team must underwent training for laboratory safety. The main topics of the training program includes:
- Laboratory & Personal Protective Equipment
- Operation Procedure for Handling biological material
- Operation Procedure for Handling Toxic Chemicals
- Operation Procedure for Emergency Situation
Our training program was based on Safety Guidelines for Biosafety Level 1 to Level 3 released by Centers for Disease Control (CDC).
Safe Design
A. Chassis Organism
We use two strains of Escherichia coli as chassis organism for our Project, one is “E. coli str. K-12. MG1655” and the other is “E. coli DH5a”. Both of them are not pathogen for human beings therefore are totally safe and are handled under guideline for biosafety.
B. New Part
In our project, we introduce three new protein for our nitrate regulation pathway: nitrite reductase (BBa_K2275001), glutamate dehydrogenase (BBa_K2275002), and glutamine synthetase (BBa_K2275003). They were originally expressed by E. coli K-12, pseudomonas putida, and pseudomonas aeruginosa PAO1, all of them are safety level 1 organism and therefore being harmless to human beings.
C. Device
- Sensing Boat
- Regulation Box
The sensing well (in which we settle sensing E. coli) in our boat is designed to be sealed and disposable, and our boat is waterproof (of course), so the E. coli will never escape from our sensing boat as long as the user reclaim them properly.
We use E. coli with three construction (nitrate reductase, glutamate dehydrogenase, glutamine) for our nitrate regulation purpose. In order to prevent our regulation E. coli from escaping, we put a filter on both side of the regulation box, and also use a UV light for sterilization before we dispose the water out of our box.
Biosafety Test
To prove that the filter and UV light work successfully, we conduct a simple experiment. First, we put some E. coli into our device, settle the filter and UV light at the right place, and then turn on our device. We took some water sample from the exit of our device, which supposed to be discharge into the fish pond in our project, and then apply the water sample on the agar plate. The number of the patch showed on the plate will tell us how efficient our safety design is.
Twelve hours later, the plate was totally clean and no bacteria patch showed on it. On the other hand, there are lot of patches appearing on the control plate (with no filter nor UV light). This result proves that our biosafety design works successfully and has the ability to prevent our E. coli from escaping.
