Team:Botchan Lab Tokyo/Safety
Botchan Lab. Tokyo
Safety
Safety
Safe Project Design
Describe the goal of your project: what is your engineered organism supposed
to do? Please include specific technical details names of important parts.
-Our bacteria will be engineered to be extremely resistant to mutations caused by many
factors such as IR (ionizing radiation), MMC (mitomycin), ROS (reactive oxygen species),
etc. This resistance will be acquired by their incredible ability of DNA repair systems
and oxidative stress tolerance. If we combine this E.coli with other technologies,
we can develop innovative solutions to serious problems in Fukushima prefecture, radioactive
contamination area. High DNA repair ability can also be used to improve cloning technology (in Vivo E.coli Cloning).
Which whole organisms, including viruses and cell lines, are you planning to use or using in your project?
-We are planning to use Escherichia coli DH5a (alfa) strain as a host organism.
We are planning to use Deinococcus radiodurans NBRC 15346 strain as a gene-donating organism.
We will take RecA, PprM, PqqE genes from D.radiodurans and introduce them in the host cell.
Source Link
https://www.nite.go.jp/nbrc/catalogue/NBRCCatalogueDetailServlet?ID=NBRC&CAT=00015346What risks could these organisms pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab?
-Nothing. Our bacteria must not survive outside the laboratory.
What experiments will you do with your organisms and parts?
-Our bacteria is meant to be extremely resistant to many mutation factors such as
IR, ROS, MMC. So, we will test whether our bacteria is able to survive under the mutagens exsisting conditions.
And also, our bacteria is meant to synthesize very long DNA chain (such as plasmid)
at one time. So, we will introduce the fragmented DNA into the cell and check whether the fragments are
bounded each other in correct form.
Our bacteria will get these ability by introducing some DNA repair genes (RecA, PprM, PqqE) from
D.radiodurans NBRC 15346 into E.coli DH5a (alfa) cell.
What risks could arise from these experiments?
-We are planning to radiate IR which is harmful for human to our bacteria.
We will also add MMC and other cancer induced chemicals into the culture of our bacteria.
So, we have to be very careful to use such mutagens. For instance, we received the lecture about
usage of radiation. And we will use the Geiger counter during the radiation experiment. As for the MMC
experiment, we will wear rubber gloves (one of PPEs) not to touch the harmful chemical directly.
Safe Lab Work
What is the Safety Level of your Lab?
-We work in Level 1 Labs and we use only Risk Group 1 organisms.
What is the measure you have taken to reduce risk during the experiment?
*Do not eat, drink and smoke in laboratories.
*Wear lab coats inside the laboratories.
*Do not bring in things from the outside as much as possible.
*Open containers with bacteria and handle them inside the clean bench.
*Wash hands before and after experiment.
What safety training have you received?
-We have been taken safety trainings, such as lab access and rules by our PIs. We learned about biosafety levels, disinfection and sterilization, emergency procedures and transport rules in the genetic recombination experiment lecture at the university.
Please see our safety form.
https://2019.igem.org/Safety/Final_Safety_Form?team_id=3226