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Team:INSA Toulouse/contenu/safety/iGEM safety
From 2013.igem.org
iGEM Safety
Safety forms were approved on September 29, 2013 by the iGEM Safety Committee.
Chassis Organism
| Species | Strain n°/name | Risk Group | Risk group source link | Disease risk to humans? If so, which disease? |
| E.coli (K12) | DH5-alpha | 1 | Link | Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. |
| E. coli (K12) | DH5-1 | 1 | Link | Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. |
| E. coli (K12) | MG1655 | 1 | Link | Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. |
| E. coli (K12) | BL21-DE3 | 1 | Link | Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. |
| E. coli (K12) | XL1-Blue | 1 | Link | Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. |
| Chromobacterium violaceum | CV026 | 2 | Link | Yes. Pathogen for humans and vertebrates, but normally no transmission between the two host groups. It rarely infects humans, but when it does it causes skin lesions, sepsis, and liver abscesses that may be fatal. |
New and/or Modified Coding Regions
| Part number | Description | Where did we get the physical DNA for this part | What species does this part originally come from? | What is the Risk Group of the species? | What is the function of this part, in its parent species? |
| BBa_K1132000 | T7 Polymerase | PCR extraction from genomic DNA of E. coli K12 BL21-DE3 | Bacteriophage T7 | 1 | RNA Polymerase |
| BBa_K1132001 | Tp901 sites Bxb1 sites pT7 BBa_B0015 Bba_J61048 |
Gene synthesis (GeneCust) | Bacteriophage TP901 Bacteriophage BXB1 Artificial sequence (Bonnet et al. 2013) Double terminator T1 terminator |
1 | Recombinase site Recombinase site Promoter region Transcription terminator Transcription terminator |
| BBa_K1132002 | Tp901 sites Bxb1 sites pT7 BBa_B0015 |
Gene synthesis (GeneCust) | Bacteriophage TP901 Bacteriophage BXB1 Artificial sequence (Bonnet et al. 2013) Double terminator |
1 | Recombinase site Recombinase site Promoter region Transcription terminator |
| BBa_K1132003 | FimE sites PhiC31 sites BBa_I712074 |
Gene synthesis (GeneCust) | E. coli (K12) FimE Bacteriophage PhiC31 T7 Promoter |
1 | Recombinase site Recombinase site Promoter region |
| BBa_K1132004 | FimE sites PhiC31 sites BBa_I712074 |
Gene synthesis (GeneCust) | E. coli (K12) FimE Bacteriophage PhiC31 T7 Promoter |
1 | Recombinase site Recombinase site Promoter region |
| BBa_K1132005 | BBa_R0040 riboregulator R0.1 BBa_B0015 BBa_R0065 riboregulator R0.2 |
Gene synthesis (GeneCust) | TetR promoter Artificial sequence Double terminator Promoter (lambda cI and luxR regulated) Artificial sequence |
1 | Promoter region RNA sequence for regulation Transcription terminator Promoter region RNA sequence for regulation |
| BBa_K1132006 | BBa_R0040 riboregulator R1.1 BBa_B0015 BBa_R0065 riboregulator R1.2 |
Gene synthesis (GeneCust) | TetR promoter Artificial sequence Double terminator Promoter (lambda cI and luxR regulated) Artificial sequence |
1 | Promoter region RNA sequence for regulation Transcription terminator Promoter region RNA sequence for regulation |
| BBa_K1132007 | BBa_R0040 riboregulator R2.1 BBa_B0015 BBa_K592006 riboregulator R2.2 |
Gene synthesis (GeneCust) | TetR promoter Artificial sequence Double terminator FixK2 promoter Artificial sequence |
1 | Promoter region RNA sequence for regulation Transcription terminator Promoter region RNA sequence for regulation |
| BBa_K1132008 | BBa_R0040 riboregulator R4.1 BBa_B0015 BBa_R0082 riboregulator R4.2 |
Gene synthesis (GeneCust) | TetR promoter Artificial sequence Double terminator OmpR-controlled promoter Artificial sequence |
1 | Promoter region RNA sequence for regulation Transcription terminator Promoter region RNA sequence for regulation |
| BBa_K1132009 | PhiC31 integrase | From the plasmid A24 from Suiti Laboratory (MIT) | bacteriophage phiC31 | 1 | Recombinase |
| BBa_K1132010 | Tp901 integrase | From the parts KC529324 from Jerome Bonnet | bacteriophage Tp901 | 1 | Recombinase |
Risks
The two chassis strains (E. coli, C. violaceum) are potential pathogenes for humans. For E. coli, the risks are minimal, but good laboratory practises were applied to avoid possible contaminations. For C. violaceum, all experiments involving this strain will be performed in a P2 facility and under the supervision of a member of the permanent staff who is the manager of the P2 facility.
There is no environmental risks.
C. violaceum is used only for parts characterisation and is not part of the original project. For the project itself, only the E. coli K12 is used and the potential risks of converting the project into a commercial product are already assessed for industrial purposes.
In the potential case where some parts should be used for other purposes than research, precautions shoud be taken to assess the safety of these microorganisms. No parts will be introduced into C. violaceum, avoiding possible antibiotic resistance of the strain.
All students receive a 2 hours safety course provided by the biosafety commitee executive. All of them were required to take a test at the end, and all of them passed.
There is an institutional biosafety commitee. The project was approved by several teachers of the institute.
The Biosafety level of our lab is 2. The institute has a P2 room fully equiped for manipulating level 2 risk group organisms.
The chassis organism risk group is 1. However, we also use a level 2 test chassis that is only manipulated in the P2 facility.
