Difference between revisions of "Team:Goettingen/Safety"
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<p>We aim to create an engineered <em>E. coli</em> which is capable of producing a range of proteins and enzymes, which will, once purified, assemble automatically to form one large protein complex, the Flexosome. The Flexosome consists of a scaffoldin and dockerins connected to enzymes. The aim is to be able to connect any enzyme of choice to the construct, therefore focusing the enzymatic activity in one place. This can be then applied to, i.e. enzymatic cascades. Due to the localized reactions their efficiency may be increased and reaction times shortened. The Flexosome may be applied to any kind of process ranging from industrial and pharmaceutical purification to breakdown of unwanted substances in the environment.</p> | <p>We aim to create an engineered <em>E. coli</em> which is capable of producing a range of proteins and enzymes, which will, once purified, assemble automatically to form one large protein complex, the Flexosome. The Flexosome consists of a scaffoldin and dockerins connected to enzymes. The aim is to be able to connect any enzyme of choice to the construct, therefore focusing the enzymatic activity in one place. This can be then applied to, i.e. enzymatic cascades. Due to the localized reactions their efficiency may be increased and reaction times shortened. The Flexosome may be applied to any kind of process ranging from industrial and pharmaceutical purification to breakdown of unwanted substances in the environment.</p> | ||
<p>Right now the team is working on a proof of concept. Our first Flexosome will contain phosphatase, esterase and cellulase found in metagenomic libraries along with RFP, a fluorescent reporter gene. Once our <em>E. coli</em> is able to express and produce this functional Flexosome, this will open the door for more complex and specific applications.</p> | <p>Right now the team is working on a proof of concept. Our first Flexosome will contain phosphatase, esterase and cellulase found in metagenomic libraries along with RFP, a fluorescent reporter gene. Once our <em>E. coli</em> is able to express and produce this functional Flexosome, this will open the door for more complex and specific applications.</p> | ||
Revision as of 03:09, 19 September 2015
Safety in iGEM
[[File:Team_Goettingen2015.JPG|right|300px|thumb]]We aim to create an engineered E. coli which is capable of producing a range of proteins and enzymes, which will, once purified, assemble automatically to form one large protein complex, the Flexosome. The Flexosome consists of a scaffoldin and dockerins connected to enzymes. The aim is to be able to connect any enzyme of choice to the construct, therefore focusing the enzymatic activity in one place. This can be then applied to, i.e. enzymatic cascades. Due to the localized reactions their efficiency may be increased and reaction times shortened. The Flexosome may be applied to any kind of process ranging from industrial and pharmaceutical purification to breakdown of unwanted substances in the environment.
Right now the team is working on a proof of concept. Our first Flexosome will contain phosphatase, esterase and cellulase found in metagenomic libraries along with RFP, a fluorescent reporter gene. Once our E. coli is able to express and produce this functional Flexosome, this will open the door for more complex and specific applications.
In this safety page, we divided our safety procedure into 4 general parts, which are:
1. Safety Rules in Laboratory
2. Safe Project
3. Safety laws and regulations in Germany
4. Safe packaging and customs
1. Safety Rules in Laboratory
Before working in the lab all team members were informed in a safety instruction about the right behavior in the lab:
- S1 laboratory safety measures and practices
- Wearing safety clothes (lab coat, rubber gloves, safety glasses and safety masks)
- Fire safety regulations and emergency action
- Accident prevention
- Documentation of experimental work and troubleshooting
- Chemical safety regulations and hazards
- Handling synthetic biology waste
Furthermore the following topics were covered:
- Basic information about iGEM and synthetic biology
- General methods and techniques needed for DNA isolation cutting, joining of DNA and plasmid transformation
In order to minimize contamination risks, we do the experiments in laminar air flow (clean bench). Furthermore work areas like the open bench for no contamination relevant experiments as well as the chemical fume hood for critical chemicals were used.
All laboratory activities were done by the students under supervision of the instructors.
2. Safe Project
Prof. Rolf Daniel is responsible for the safety in our lab. The guidelines follow Ordinance on Safety and Health Protection at Workplaces Involving Biological Agents Biological Agents Ordinance - BioStoffV .
We did not use organisms from Risk Group 3 or 4, and did not in no kind of way released genetically modified organisms, or their products, into the wild.
Like the huge majority of iGEM teams we were working only with Risk Group 1 organisms, such as E. coli TOP10 and E. coli BL21. From these no special toxins or virulence factors are known that could be harmful to humans or require any additional safety precautions. For Risk Group information on Escherichia coli K-12 (and derivatives: DH5alpha, TOP10, etc.) see DSMZ.
In the beginning of the project Acetivibrio cellulolyticus, Clostridium thermocellum, Clostridoum cellulolyticum and Bacteroides cellulosolvens were cultured and their DNA was used to amplify specific dockerins by PCR. After this step E. coli was the only organism handled in the lab.
With our project we are producing synthetic biological waste. Agar plates, pipet tips, falcons, tissue, rubber gloves or any other contaminated waste is collected in special S1 waste containers and decontaminated with the autoclave before disposal.
Old cell cultures such as liquid wastes in general are also decontaminated by autoclaving according to the laboratory standard operational procedures and afterwards disposed to the sanitary sewer system.
We use lab coats, gloves and safety glasses in order to minimize contamination risks while working. Safety clothes are only worn in the lab and taken off when leaving the laboratory environment.
3. Safety laws and regulations in Germany
Federal Office of Consumer Protection and Food Safety
Gesetz zur Regelung der Gentechnik (Gentechnikgesetz - GenTG)
4. Packaging and Customs
When submitting our parts for the 2015 competition, we used the standard DNA Submission Kit that came with the 2015 Distribution, and followed the directions on the Registry .