Team:Northwestern/Growth

From 2014.igem.org

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Notes on culturing

Nitrosomonas europaea

Because this chemolithoautotroph metabolizes ammonia as its primary energy source, this bacterium strain covers a large range of substrates useful for bioremediation, waste management, and possible pollutant degradation. In addition, Nitrosomonas is nitrogen oxidizing and as a result, can limit carbon dioxide fixation in plants.

  • obtained starter culture from Freddy Chen from George Wells's Lab in the Environmental Engineering department
  • it is grown in the dark, at 30C, in a special buffer called NE buffer (has specific concentrations of ions)
  • instead of measuring its OD, (researchers have not been able to culture it past 0.1), we monitored growth by pH
  • the starting pH should be around 7.5, after about 21 days, the pH will drop to about 6, at which point the culture will need to be split into new flasks
Streptomyces rimosus

Streptomyces is a bacterium that naturally produces key antibiotics used on a daily basis in synthetic biology (such as chloramphenicol and neomycin). This particular strain is one of the best industrially characterized strains for the purpose of antibiotic production. Better understanding its genetic compatibility may help with antibiotic resistance.

  • obtained starter culture from the Kelleher Lab in oncobiology research
  • it is grown in ISP-2 at 30C, in liquid and on plates
  • optimal pH for growth is around 7-7.2
Pseudomonas fluorescens

Pseudomonas fluorescence is a lesser known strain of bacteria that naturally produces an antibiotic called mupirocin, which can be used for the commercial purpose of treating ear, skin, or even eye disorders. From a more industrial aspect, this bacterium aids crop growth for agricultural purposes by providing protection from environmental pathogens, infections, and/or toxic pollutants.

  • Starter culture from the Wang Lab under the Department of Chemical and Environmental Engineering
  • cultured at 30 celsius
Environmental Strains

Collection of several water samples from Lake Michigan from Northwestern's lakefill provided two additional environmental strains. Characterization of these strains were done in the same cell free platform. The two strains collected were Comammonas testostrene and Enterobacter aerogenes. Comammonas testostrene is a strain of bacterium that plays a big role in bioaugmentation. Characterization of this strain could significantly expedite the process of biowaste degradation in the near future. Successful characterization of our part in these different lysates would also prove that our platform can be universally utilized.

  • grown in standard LB at 30C
  • continued to grow at RT (room temperature) and daily dilutions were done per day to further isolate our samples
  • restreaked daily to isolate selective homogenous colonies