Team:Bielefeld-CeBiTec/Organisms

iGEM Bielefeld 2015


Organisms

The favorite pets of a synthetic biologist

Escherichia coli

We used the following E. coli strains during our project:

  • KRX (Source: Promega)
    • Genotype: [F´, traD36, ΔompP, proA+B+, lacIq, Δ(lacZ)M15] ΔompT, endA1, recA1, gyrA96 (Nalr), thi-1, hsdR17 (rk–, mk+), e14– (McrA–), relA1, supE44, Δ(lac-proAB), Δ(rhaBAD)::T7 RNA polymerase.
  • ER2566 (T7 Express Competent E. coli (High Efficiency), source: New England Biolabs)
    • Genotype: F- λ- fhuA2 [lon] ompT lacZ::T7 gene 1 gal sulA11 Δ(mcrC-mrr)114::IS10 R(mcr-73::miniTn10-TetS)2 R(zgb-210::Tn10)(TetS) endA1 [dcm]

Some other strains were used just once to investigate their performance in in vitro cell-free protein synthesis:

  • Rosetta-gami 2 (Source: Novagen, part of Merck KGaA)
    • Genotype: Δ(ara-leu)7697 ΔlacX74 ΔphoA PvuII phoR araD139 ahpC galE galK rpsL F′[lac+ lacIq pro] gor522::Tn10 trxB pRARE2 (CamR, StrR, TetR)
  • BL21 (DE3) (Source: New England Biolabs)
    • Genotype: fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5

Bacillus subtilis

In our project we used B. subtilis strain 168 with genotype trpC2 for the GABA operon sequence. You can find further information about the origin of this strain in Zeigler et al.

Agrobacterium tumefaciens

We isolated the blc-operon from A. tumefaciens C58.1. For detailed information about this organism please be referred to Wood et al. 2001.

References

Wood, D. W.; Setubal, J. C.; Kaul, R.; Monks, D. E.; Kitajima, J. P.; Okura, V. K. et al. (2001): The genome of the natural genetic engineer Agrobacterium tumefaciens C58. In: Science (New York, N.Y.) 294 (5550), S. 2317–2323. DOI: 10.1126/science.1066804.

Zeigler, Daniel R.; Prágai, Zoltán; Rodriguez, Sabrina; Chevreux, Bastien; Muffler, Andrea; Albert, Thomas et al. (2008): The origins of 168, W23, and other Bacillus subtilis legacy strains. In: Journal of bacteriology 190 (21), S. 6983–6995. DOI: 10.1128/JB.00722-08.