Difference between revisions of "Team:HokkaidoU Japan/Safety"
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Revision as of 19:41, 1 November 2017
Safety
Safe Project Design
- We used E. coli TOP10 Competent Cells, which is BSL-1 and non-pathogenic.
- All the proteins we used do no harm to humans/ animals/ plants.
- We analyzed all the proteins after extracting from E. coli.
Safe Lab Work
- We conducted all the experiments in Safety Level 1 Lab.
- 8 members took a safe instruction for Gene Modification at Hokkaido University and shared it to everyone.
- Every members obeyed the rule of the laboratory, and asked our professor how to do when one first do an experiment.
The chassis organism we are using for this project.
- E. coli (K 12) TOP10
Highest Risk Group Listed
Risk Group 1This is a list of our new coding regions in our projects.
| Part number | Source of DNA | Species | Risk group | Function |
|---|---|---|---|---|
| BBa_K2015000 | IDT gBlock DNA synthesis | E. coli | 1 | Self-assembling region (SAR) for the circularization of proteins |
| BBa_K2015001 | IDT gBlock DNA synthesis | E. coli | 1 | One of self-assembling peptides |
| BBa_K2015002 | IDT gBlock DNA synthesis | E. coli | 1 | One of self-assembling peptides |
| BBa_K2015003 | IDT gBlock DNA synthesis | E. coli | 1 | G7 linker containing 7 Glycine |
| BBa_K2015004 | IDT gBlock DNA synthesis | E. coli | 1 | This part can form disulfide bonds between Sulfur residues for multimerization of protein |
| BBa_K2015005 | IDT gBlock DNA synthesis | E. coli | 1 | This part can form disulfide bonds between Sulfur residues for circularization of protein |
| BBa_K2015008 | IDT gBlock DNA synthesis | E. coli | 1 | This part can be formed multimerized protein by using BBa_K2015002 |
| BBa_K2015009 | IDT gBlock DNA synthesis | E. coli | 1 | This part can be formed multimerized protein by using BBa_K2015001 |
| BBa_K2015010 | IDT gBlock DNA synthesis | E. coli | 1 | CDS for circularized protein |
| BBa_K2015011 | IDT gBlock DNA synthesis | E. coli | 1 | LacI expression unit |
| BBa_K2015012 | IDT gBlock DNA synthesis | E. coli | 1 | The sequences of downstream can be induced strictly by IPTG |
| BBa_K2015013 | IDT gBlock DNA synthesis | E. coli | 1 | A part of circularized GFP |
Description of the biological materials we are using in the lab.
Risks to the safety and health of team members or others working in the lab.
Some materials possess risks to team members. For example, Ethidium Bromide is an intercalating agent so it must be used by with personal safety gear. All lab staffs were trained according to safety manual provided by Hokkaido University, to prevent risks. We took on ourselves to compile a shortlist of often used dangerous materials and safety procedures in our project.Dangerous chemicals
- Chloroform
- corrosive and toxic : must be used in fume hood
- Ethidium Bromide
- intercalating agent : must be used with personal safety gear
- Ethanol
- flammable : must not be used near open flame or in large quantities
- Liquid Nitrogen
- cryogenic container and cryogenic gloves must be used
Procedures and equipment
- Agarose gel production
- heating in sealed container (rupture risk), scalding hot and vicious during preparation (burn injury risk) - remove container lid before heating in microwave, use safety gear, wait for a few moments before removing from microwave
- Benson burner
- fire risk: DO NOT use flammable materials especially ethanol near open fire
- Centrifuge
- high velocity: balance appropriately, observe the machine till it reaches top velocity
- Autoclave
- high pressure: check the water level, DO NOT open when pressurized
- UV radiation
- damage to eyes and skin: use glove and UV box or UV shield
Non-pathogenic bacteria (policy requires treating as pathogenic, as precaution)
- DH5α
Safety equipment
- Gloves
- Coats
- Goggles
- UV Box
- UV shield
- Clean bench
Waste disposal and sterilization
- All equipment and wastes coming in contact with bacterial were sterilized by autoclave or bleach.
- All chemical compounds were disposed according to requirements for their disposal.
- Table surface used for work were sterilized with 70% ethanol before and after a procedure.
Chemical Usage
All chemical compounds were used according to their manuals and respective material safety data sheets.Genetic material
All genes used in this project come from non-pathogenic bacterial strains of E. coli. Expressed proteins did not show any toxic effect to their host. Our biobricks do not have any foreseeable selective advantage if released to the environment. After consideration we could not find any usage pausing a security concern.Risks to the safety and health of the general public, if released by design or by accident.
Some materials pose risks to the general public. For example, Ethanol is a flammable solution so it must not be used by open fire. Not to release those materials, all lab staff is trained according to safety manual provided by Hokkaido University.Risks to the environment, if released by design or by accident.
The E. coli strains we use in our lab, are saved in our lab. As a precaution all materials coming in contact are sterilized before and after. Reference Federal Register, (1986) Vol. V1: 88, 6952-16985.Risks to security through malicious misuse by individuals, groups, or countries.
There is no foreseeable risk in misuse of our generated genetic material. Our generated genetic material performs basic functions in biology. However, it is impossible to guard against the incorporation of our parts in malicious settings.Risks which might arise when our project move from a small-scale lab study to become widely used as a commercial/industrial product.
Our project is about optimizing the expression of genes. Our device does not contain a coding site. Therefore, risk will arise when other users assemble our parts with dangerous coding sites. We have to caution the users when assembling with dangerous coding sites.Design features to address safety risks.
Our device only contains sequences that regulate the expression of genes (Promoter, RBS, and terminator). Therefore, our device in itself does not contain any safety risks and does not have design feature to address safety risks.Safety training we received.
8 members of our team received a lecture class regarding gene recombination that was held in Hokkaido University, and shared the information with the whole team. It is based on 'Act on the Conservation and Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified Organisms'.Biosafety provisions
Link to our institution biosafety guidelines.
http://www.hokudai.ac.jp/jimuk/reiki/reiki_honbun/u010RG00000583.htmlOur Institutional Biosafety Committee.
We have a permission to engage in the experiments from the safety officer of genetic recombination of Hokkaido University. All members participating in the experiments are registered with this office. All members are trained according to the safety demands of safety officer of genetic recombination.Our country's national biosafety regulations and guidelines
Japan is participating in cartagena act.Biosafety Level rating of our lab.
Our lab's Bio safety level is 2.The Risk Group of our chassis organisms.
The Risk Group of our chassis organisms is 1.Faculty Advisor
Sone TeruoTamazaki Ken-ichi
