Team:Mingdao/Safety

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• About Us
• Team Members
• Attribution

• Motivation
• Design
• Experiment
• Proof
• Demo
• Result
• Notebook

• Basic Parts
• Composite Parts
• Parts Collection

• Introduction
• Principal
• Experiment
• Future
• Interlude

• Lab Work
• Project Work
• Shipment
• Q&A

• Survey
• Promotion

• Bronze
• Silver
• Gold

Safety agents

• usually pose a minimal potential threat to laboratory workers and the environment
• do not cause any diseases in healthy people

Safe lab

• standard open laboratory benches
• labs are not usually isolated from the general building
• biological safety cabinet and laminar flow cabinets are used

Trained Lab Personnel

• training on the specific procedures to handle the biomaterials
• supervised by a trained microbiologist or scientist

Implemented Standard Microbiology Practices

• mechanical pipetting
• safe sharps handling
• avoidance of splashes or aerosols
• decontamination:·
  • all potentially infectious materials are decontaminated prior to disposal, generally by autoclaving
• personal hygiene
• hand washing
• prohibition on eating, drinking or smoking in the lab
• protective equipment
• eye protection·
• gloves and a lab coat·

• biohazard signs are posted and access to the lab is limited

Most of our team members have never or seldom heard of synthetic biology and iGEM before joining the team. Therefore, all the members have to participate in the safety courses held by our team instructor, Dr. Pei-Hong Chen, before all of us have permission to use the lab. The courses include Autoclave safety, Biological Material Safety, and Chemical Safety.

Bacterial strains: Escherichia coli DH5α and BL21

Escherichia coli DH5α and BL21 are recognized as non-pathogenic and disabled, and may be considered to be equivalent as the Advisory Committee on Dangerous Pathogens (A CDP) hazard group 1. They are not considered pathogenic to humans or animals. They are expected to have limited survivability in the environment and often have auxotrophic requirements, which are unlikely to be satisfied outside of laboratory culture.

According to the Centers for Disease Control and Prevention (CDC) Guidelines in Taiwan, these materials can be handled at Biological Safety Level One (BSL-1) containment. Biological Safety Level One (BSL-1) containment, using standard microbiological practices, is suitable for work involving well-characterized microbiological organisms not known to consistently cause disease in immunocompetent adult humans, and present minimal potential hazard to laboratory personnel and the environment.

Species name (including strain	Risk Group	Risk Group Source	Disease risk to humans?	Part number/name	Gene Acquired by
E. coli DH5α	1	NIH	No	BBa_K1991000/AOX1_pSB1C3	IDT DNA synthesis
E. coli DH5α	1	NIH	No	BBa_K1991001/AOX2_pSB1C3	IDT DNA synthesis
E. coli DH5α	1	NIH	No	BBa_K1991002/Pcons-RBS-AOX1_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991003/Pcons-RBS-AOX2_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991004/LO-BamHI_pSB1C3	Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991005/LO-AOX1-His_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991006/LO-AOX2-His_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991007/Pcons-RBS-LO-BamHI_pSB1C3	Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991008/Pcons-RBS-LO-AOX1-His_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991009/Pcons-RBS-LO-AOX2-His_pSB1C3	IDT DNA synthesis & Biobrick
E. coli DH5α	1	NIH	No	BBa_K1991010/Pcons-RBS-LO-GFP-His_pSB1C3	IDT DNA synthesis & Biobrick
E. coli BL21	1	NIH	No	pET vectors	Other lab

Solutions of sending DNA parts to the Registry
1.Search for and follow the regulations published by iGEM HQ

According to the rules of submission at iGEM Registry of Standard Biological Parts, we will follow the regulation for sending the parts. (1) Follow the shipping guidelines for submitting samples of the parts. Samples have been sent as mini-prepped plasmid DNA (pSB1C3). (2) Submit by using the 96 well plates, sealed completely (adhesive foil/plastic), and covered with a protective lid. (3) Provide a tracking number for the package (4) Disguised or hidden are not allowed and avoid customs confusion.

2. Ensure less risk as possible

Prepare backups for emergency situations, and check problems by consulting with local post offices.

A. Risks to the safety and health of team members or others in the lab?

E. coli lab strains (DH5α and BL21) have no known survival mechanisms outside the lab culture, and they only caused minimal harm to people or environment. Therefore, they are considered to be safe and practical for research. The only excluded condition is horizontal gene transfer. In our project, the gene products were oxidoreductase enzymes, AOX, which catalyze the oxidation of primary alcohol. The enzymes have been used in many biotech and industrial applications and supposed not to be dangerous to the health of our members.

B. Risks to the safety and health of the general public if released by design or accident?

Our genetically engineered bacteria producing alcohol oxidase enzymes would be used on the sample test strip of the Blood Alcohol Meter, an electrochemical analyzer like Blood Glucose Meter seen as household items. If released by accident, the E.coli has no survival mechanisms in the environment and cannot spread spores or reproduce in the gut of an organism, so it has low threat to the public. By design, the bacteria will be killed in the process of standard sterilization process before depositing them to the test strip. The test strip we created will not contain any live GE bacteria, therefore it won’t have risks to the safety and health of the general public.

C. Risks to environmental quality if released by design or accident?

If so, the E. coli lab strain no longer has survival fitness outside the lab. And the DNA plasmids are maintained by high concentrations of antibiotics like chloramphenicol and kanamycin in our case, thus they will disappear and be degraded in the absent of antibiotics if there’s horizontal gene transfer concern. In addition, we always killed all the materials including bacteria by standard autoclave procedure before letting them leave the lab, so it won’t cause any risk to environment quality.

D. Risks to security through malicious misuse by individuals, groups or states?

Our bacteria will only be alive in production process, it will be killed right away after placing it on the test strip. A Blood Alcohol Meter will not contain any live bacteria we produced. And the genes on the plasmid vector maintained by antibiotics are supposed not to be present for a long time in a normal environment. Moreover, the gene encodes an oxidoreductase enzyme used by the methylotrophic yeast in nature. The gene products are not harmful to human and the environment.