1) Please describe the chassis organism(s) you will be using for this project. If you will be using more than one chassis organism, provide information on each of them:

Species: E. coli(K-12)

*Strain: DH5alpha
*Risk Group: 1
*Risk Group Source Link: NIH Guidelines
*Disease risk to humans? If so, which disease? : Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys.

Species: E. coli(K-12)

*Strain: JM2.300
*Risk Group: 1
*Risk Group Source Link: NIH Guidelines
*Disease risk to humans? If so, which disease? : Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys.

Species: E. coli(K-12)

*Strain: MG1655
*Risk Group: 1
*Risk Group Source Link: NIH Guidelines
*Disease risk to humans? If so, which disease? : Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys.

Species: E. coli(K-12)

*Strain: AJW678
*Risk Group: 1
*Risk Group Source Link: NIH Guidelines
*Disease risk to humans? If so, which disease? : Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys.

Species: E. coli(K-12)

*Strain: W3110
*Risk Group: 1
*Risk Group Source Link: NIH Guidelines
*Disease risk to humans? If so, which disease? : Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys.

2) Highest Risk Group Listed:

1

3) List and describe all new or modified coding regions you will be using in your project.

Part name: BBa_K1632000-7, BBa_K1632030-3

* Where did you get the physical DNA for this part (which lab, synthesis company, etc): Gene synthesis by eurofins.
* What species does this part originally come from?: E. coli (K-12)
* What is the Risk Group of the species?: 1
* What is the function of this part, in its parent species?: Inverts with recombinase

Part name: BBa_K1632010-9

* Where did you get the physical DNA for this part (which lab, synthesis company, etc): Kiga Lab.
* What species does this part originally come from?: E. coli (K-12)
* What is the Risk Group of the species?: 1
* What is the function of this part, in its parent species?: Produces fim recombinase.

Part name: BBa_K1632020-3

* Where did you get the physical DNA for this part (which lab, synthesis company, etc): Gene synthesis by eurofins.
* What species does this part originally come from?: E. coli (K-12)
* What is the Risk Group of the species?: 1
* What is the function of this part, in its parent species?: Produces chloramphenicol acetyltransferase.

4) Do the biological materials used in your lab work pose any of the following risks? Please describe.

a. Risks to the safety and health of team members or others working in the lab?
 - Without drinking large amount of sample, there is no risk. Such taking might cause irritation to skin, eyes, and respiratory tract.

b. Risks to the safety and health of the general public, if released by design or by accident?
 - Without drinking large amount of sample, there is no risk. Such taking might cause irritation to skin, eyes, and respiratory tract.

c. Risks to the environment, if released by design or by accident?
 - Production of AHL might lead to perturbation communications in wild bacteria.

d. Risks to security through malicious misuse by individuals, groups, or countries?
 - Use of plant hormone in the opposite direction could disrupt growth of crops. Malicious usage of phage might lead to perturbation of intestinal flora.

e. What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)
 - Do not eat, drink, smoke, prepare or store foods in laboratories.
 - Wear lab coats inside the laboratories.
 - Wear rubber gloves inside the laboratories.
 - Keep the laboratories tidy and clean.
 - Separate the rooms handling bacteria and the rooms that don’t.
 - Put labels on containers.
 - Only open containers with bacteria, inside the clean bench.
 - Sterilize the waste before they are thrown away.
 - Practice disinfection, sterilization, and washing hands.

5) What safety training have you received

a. Have your team members received any safety training?
 Team members have received safety training and taken classes for bioengineering including recombinant DNA technology.

b. Please briefly describe the topics that you learned about (or will learn about) in your safety training.
 Genetically modified organisms are prohibited to be taken out from the lab. So, we must always sterilize these organisms using autoclave, before disposing them.

6) Under what biosafety provisions will/do you work?

a. Who is responsible for the safety of biology labs at your institution? What are the guidelines for laboratory biosafety? Please give a link to these guidelines, or briefly describe them if you cannot give a link.
 http://www.rpd.titech.ac.jp/rpdiv/somu/cat71/detail_216.html

b. In your country / region, what are the laws and regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.
 http://www.lifescience.mext.go.jp/bioethics/anzen.html

c. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.)
 BSL1

d. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.
 Group1. It matches the rating of our lab.

Faculty Advisor Name:

Daisuke Kiga