User:Christine Doan/Notebook/Synthetic Biology/2014/03/21

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Relevant BioBricks

  • Nitrate reporter: PyeaR - GFP composite (BBa_K381001)[1]
    • Designed by Katharine Coyte, iGEM 2010 BCCS-Bristol
    • GFP coding device
    • Strong RBS
    • PyeaR is normally repressed by NsrR, protein native in most E. coli
    • When nitrates or nitrites enter, it is converted to nitric oxide, which binds to the NsrR, halting the repression and producing GFP
    • Senses nitrates within a range of 0-20mM (calculated from Miller assays
  • Nitrite reductase device: (BBa_K1196018)
    • Uses ferrocytochrome c to convert nitric oxide and water to nitrite and protons

Previous iGEM Nitrate Biosensor Projects

  • UTP-Panama: The RENBO: Thermo (2011)
    • Goal: extend range of optimal functioning for temperatures below 20C for use in highland crops, such as coffee and fruits
    • "The original BioBrick that senses the levels of nitrates in soil developed by Bristol iGEM Team 2010, was designed to operate at room temperatures, especially for 37 ° C using the PyeaR BioBricks promoter"
    • AOX (alternative oxidase) enzyme, used in the sacred lotus for thermogenesis, generates an increase in periplasmic temperature
      • Used with the HybB promoter for cold shock and OmpA as a signal peptide
      • Developed new device by combining the AOX periplasmic heat generator (BioBrick BBa_K410000) and Nitrates biosensor (BioBrick BBa_K381001) to create "The Renbo"
      • 20-37C HybB promoter expressed
      • 37C HybB promoter ceases expression
    • Future direction: exchange the HybB promoter for the CspA promoter (BioBrick BBa_K328001) to produce anti-freeze protein that prevents ice crystal formation in the cell
    • Parts used:
  • AHUT-China: Shining Purifier (2011)
    • Goal: design a wastewater cleansing system to efficiently absorb pollutants and transform them to light
    • Large amounts of sewage from petrochemical engineering and agriculture leads to inorganic nitrogen and red tide
    • Current method is anaerobic ammonium oxidation bacteria (anammox), which converts fomite to nitrogen, but it's time-consuming and oxygen-sensitive
    • Synthesize a biofilm in 4-6 days
    • Signal factor (a) prompts bacteria to stretch out flagellum, (b) release mucus, and (c) promote reproduction
      • Increased surface area = decreased release of signal factors
      • No interaction among the signal factors
    • Parts used:
      • Fluorescent protein device (BBa_K1196016)
      • Nitrite reductase device (BBa_K1196018)
      • Hydrazine synthetase device (BBa_K1196019)
      • Hydrazine oxidoreductase device (BBa_K1196020)
      • Formate dehydrogenase device (BBa_K1196021)
      • NADH:ubiquonone reductase device (BBa_K1196022)
      • Cytochrome c reducase device (BBa_K1196023)
  • Calgary: Detect and Destroy: Building FRED and OSCAR (2012)
    • Convert toxins into useable hydrocarbons
    • Tailings ponds contain byproducts of the bitumen extraction process of oil sands
    • FRED: Functional Robust Electrochemical Detector
      • Multiple output biosensor
    • OSCAR: Optimized System for Carboxylic Acid Remediation
      • Designed to target naphthenic acids (carboxylic acid-containing compounds), catechol, and nitrogen and sulfur containing heterocyclic compounds)
    • Parts used:
      • PetroBrick (BBa_K590025)
      • xylE (BBa_J33204)

Biosensor Database [2]