Safety

"Safety is not just a slogan, it is a way of life"

Summary :

iGEM Safety
Chassis Organism
New and/or modified coding region
Safety in our lab

iGEM Safety
Our safety form was approved on September X, 2014 by the iGEM team. We filled in with the help of Nathalie Doubrovine, safety officer at the LISBP.

Species name (including strain)	Risk group	Risk Group Source	Disease risk to humans ?
Escherichia coli MC1061	1	DSMZ	No. These organisms do not cause disease in healthy adult humans. (However, they might cause diseases in young children, elderly people, or people with immune system deficiencies.)
Bacillus subtilis 168	1	DSMZ	No. These organisms do not cause disease in healthy adult humans. (However, they might cause diseases in young children, elderly people, or people with immune system deficiencies.)
Aspergillus brasiliensis 246.65 CBS	1	DSMZ	No. These organisms do not cause disease in healthy adult humans. (However, they might cause diseases in young children, elderly people, or people with immune system deficiencies.)
Chaetomium globosum 148.51 CBS	1	DSMZ	No. These organisms do not cause disease in healthy adult humans. (However, they might cause diseases in young children, elderly people, or people with immune system deficiencies.)

Part number/name	Natural function of part	How did you acquire it?	How will you use it?
RBS (K090505) + Chimiotaxis + Terminator(B0015)	Chimiotaxis code for a chimeric protein (based on McpA (1-34/283-661) from B.sub and VCD_00306(28-315) from V.cholerae)	We order this gene from a synthesis company (Eurofins)	This part allows the bacterium to reach the fungus
RBS (K090505) + binding + Terminator(B0015)	"Binding" code for a chimeric binding protein (based on LYCT (1-318) from B.sub and GBPA (324-384) from V.Cholerae) with a linker between.	We order this gene from a synthesis company (Eurofins)	This part allows the bacterium to fix the fungus
RBS (K090505) + GAFP1	GAFP1 code for a antifungal. GAFP1 come from arachis hypogaea (Peanut)	We order this gene from a synthesis company (Eurofins)	This part produces a fungicide. By this way, the bacteria is able to kill fungus.
RBS (K090505) + D4E1 + Terminator(B0015)	D4E1 code for a antifungal. DAE1 is a synthetic gene	We order this gene from a synthesis company (Eurofins)	This part produces a fungicide. By this way, the bacteria is able to kill fungus.
Promoter(K823003) + (K1364000: RBS (K090505) + Chimiotaxis + Terminator(B0015))	Promoter Pveg, and Chimiotaxis part	We made it	This part allows the bacterium to reach the fungus
Promoter(K823003) + (K1364001: RBS (K090505) + Binding + Terminator(B0015))	Promoter Pveg, and binding part	We made it	This part allow the bacterium to fix the fungus
(K1364004: Promoter(K823003) + RBS (K090505) + Chimiotaxis + Terminator(B0015))+(K1364005: Promoter(K823003) + RBS (K090505) + Binding + Terminator(B0015))	Double expression cassette	We made it	This part allows the bacterium to reach and bind the fungus
Promoteur(K823003) + (K1364002 : RBS (K090505) + GAFP1)	Promoter Pveg, and GAFP1 part	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus
(K1364002 : Promoteur(K823003) + RBS (K090505) + GAFP1) + Terminator (B0015)	Promoter Pveg, and GAFP1 part with a double terminator	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus
Promoter(K823003) + (K1364003 : RBS (K090505) + D4E1 + Terminator(B0015))	Promoter(K823003) + (K1364003 : RBS (K090505) + D4E1 + Terminator(B0015))	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus
K71364010: Promoter + RBS + EcAMP) + terminator(B0015)	Promoter Pveg, RBS SpoVG, Antifungal EcAMP, double terminator	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus
K1364002 : RBS + GAFP1 ) + (K1364003: RBS + D4E1 + terminator)	RBS consensus, Antifungal GAFP1, RBS consensus, Antifungal D4E1, double terminator	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus
Promoter(K823003)+(K1364012 : RBS + GAFP1 + RBS + D4E1 + terminator)	Promoter Pveg, RBS consensus, Antifungal GAFP1, RBS consensus, Antifungal D4E1, double terminator	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus.
(K71364010: Promoter + RBS + EcAMP) + (K1364012 : RBS + GAFP1 + RBS + D4E1 + terminator)	Promoter Pveg, RBS SpoVG, Antifungal EcAMP, RBS consensus, Antifungal GAFP1, RBS consensus, Antifungal D4E1, double terminator	We made it	This part produces a fungicide. By this way, the bacterium is able to kill fungus.

Ethidium bromide
All along our project, we use non-pathogenic strains and non-toxic product, except the EtBr. We have a dark room dedicated to the use of EtBr and UV. This room is key-closed and everyone entering into the room must wear gloves, glasses and lab coat. Everything which is in contact directly with something in this room has to stay here. Two specific garbages are dedicated to the gloves and the agarose gels contaminated

Basic safety principles
All along our project, we use non-pathogenic strains and non-toxic product, except the EtBr. However, we have to apply the basic safety principles into a laboratory room: - It is forbidden to smoke in all the INSA's rooms. - It is forbidden to drink and eat in the laboratory rooms. - It is compulsory to wear a closed lab coat in cotton. - It is compulsory to wear closed shoes. - Long hair must be tied back - Oral pipetting of any substance is prohibited in any laboratory

"Precaution is better than cure"

Basic safety principles
There is also others precaution when working with biological organisms. - We need to wash our hands regularly. - It is compulsory to wear gloves except when working with an electric burner. - In some case (UV light, projection risk), it is compulsory to wear protection glasses. Indeed, different machine are used to keep a sterile area.

"If you don't think it's safe, it's probably isn't"

iGEM Toulouse INSA Toulouse 135, Avenue de Rangueil 31400 TOULOUSE e-mail: igemtoulouse2014@gmail.com