User:Jonathan Cline/Notebook/Melaminometer

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http://upload.wikimedia.org/wikipedia/commons/thumb/0/05/Melamine.svg/150px-Melamine.svg.png Melamine C3H6N6


For biologists: Read the "Much needed product for world health" proposal first, then skip to the "Specific Metabolic Activity".

For DIY-biologists or iGEM types: Read the "As proof of DIY Bio" proposal first, then skip to "Biochemistry Background".

For lay people: Read the "Much needed product for world health" proposal first, then skip to "Toxicology Background" and other background information.


Project Proposal

Product Need

  • Commercial food testing is unlikely to change in the short term
  • Commercial testing has failed to meet safety standards demanded by consumers
  • A method for establishing harmful content in food is needed at "Point-of-care"
    • Harmful product may especially exist in resource-constrained communities where commercial testing is lax
    • Consumers should have a means of testing food themselves.


Background

Technical

Historical Perspective

2008: Large-scale contamination found in wide variety of milk products exported from China. Hospitalized and/or affected population reported in popular media to be between 10,000 and 50,000 people.

2007: Pet food contamination triggers intensive research into chemically or physically determining presence of harmful agents & their metabolic pathways.

Mid 2000s: Metabolism of melamine and/or related compounds in bacteria metabolism studied in further detail using genetic analysis.

Mid 1990s: biotech interest in melamine metabolism due to possibility of enhancing plant nitrogen takeup in optimizing fertilization. Not much known about metabolic pathway. Nothing known about related gene expression. Bacterial strains known; Pseudomonas isolates, two Klebsiella isolates, and a Rhodococcus isolate).

Early 1990s: Melamine catabolism studied as means to degrade atrazine herbicide in the U.S. Pseudomonas used and first plasmids created.

Mid 1980s: HPLC analysis allows quantitative measurement of melamine pathway; prior to this, identification of intermediates is difficult.

Late 1970s: Partial, tentative, data into melamine degradation is published.

Dry Lab Design Method

  1. Project definition
    1. Iteratively (re-)define project
    2. Verify theoretical feasibility
    3. Repeat from above
  2. Project Feasibility
    1. Research existing metabolism/catabolism/enzymatic activity
    2. Research existing microbe structure
    3. Verify theoretical feasibility of inserting metabolic path into desired microbe
      1. Need a suitable vector -- not all plasmids are appropriate for all gut bacteria
      2. Must be stable, i.e., reproduce from generation to generation
    4. Verify theoretical compatibility of biological components
    5. Repeat from above
  3. Project Testability
    1. Research method for testing microbe
    2. Research required controls, blanks, positives, samples
    3. Repeat from above for design of test method, as necessary
  4. Project Modeling
    1. Model theoretical operation of microbe
    2. Model theoretical compatibility of components
    3. Model testing method
    4. Obtain expert assistance/advice on design & models
    5. Repeat from above
  5. Project Implementation Scoping
    1. Scope building organism (cost, labor, designer)
    2. Funding source
    3. Materials source
    4. Proceed to wet lab phase

Wet Lab Implementation Method

  • TBD
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