Team:DTU-Denmark/Safety protocols

Safety considerations regarding the use of E.coli DHα Most E.coli strains are harmless as they are a normal part of human gut flora, but some, such as serotype O157:H7, can cause serious food poisoning in humans. 3 The genetic set up of E.coli DH5α has been engineered in such a way, that these strains can be considered as safe laboratory strains, if standard safety protocols are followed. The genetic characteristics of the DH5α strains are (1,2): F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ– The most important genetic modifications for lab security are explained in the next paragraph. F- means that the strain doesn´t have the ability to develop F-pili, so it can´t transfer genetic information by horizontal gene transfer. The strain DH5α is also recombination deficient (recA1),so the mutant gene limits recombination of the plasmid with the E.coli genome so that the plasmid inserts are more stable. At the same time RecA mutant strains are very sensitive to UV-light, because their ability to do DNA repair is impaired. Local biosafety group We have a local biosafetygroup at our institution, and they have advised us to follow standard safety protocols for genetic engineering and molecular biology that is the standard practice at our institution. None of the biobricks submitted in this project comprise any elevated danger of researcher safety, public safety, or environmental safety, if the standard practice are followed. Reference: 1. Woodcock, D.M. et al. (1989) Nucl. Acids Res., 17, 3469–3478. 2. Raleigh, E.A., Lech, K. and Brent, R. (1989). In F.M. Ausebel et al. (Eds.), Current Protocols in Molecular Biology, (p. 1.4). New York: Publishing Associates and Wiley Interscience. 3. http://en.wikipedia.org/wiki/Escherichia_coli