IGEM:IMPERIAL/2007/Projects/In-Veso/Specifications

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In-Veso Gene Expression: Specifications



Contents


The specifications outline the characteristics of the chassis being sought, and the quantities that will be varied in experiments. The range for each input variable is also detailed. All lists are prioritised in order to help planning of experiments.

Input Variables

Temperature

Range from 4-37oC with priorities at:

  1. 25oC
  2. 4oC
  3. 6-20oC
  4. 30-37oC


Media

These are the different media the technique should be tested in, in order of priority:

  1. Water
  2. X-Gal
  3. E. coli Biofilm
  4. Agar Gel
  5. Polyacrylamide
  6. Paper

Note: take into account Osmolarity of the media.


Parts

List of parts from the registry we would like to test in-veso.


Piping

How narrow a pipe, and how fast, can the vesicles be passed through without bursting? Testing range..?

This is valuable for determining the spray system to be used in the Biofilm application. It also helps with laboratory manipulations - pipetting, etc.

Dependent Quantities

Life span

  • Measured in hours

Rate of protein synthesis

  • The units will depend on the reporter being used, as well as the volumetric unit.

Rate of PoPs

  • Indirectly measured in GFP synthesis per CFU per second.

Membrane Traffic

  • List of particles that can cross the α-hemolysin pores

Resources

These are the resources that are available to us:

Characterisation Chart

The following chart cross-references the input variables to the dependent quantities. Each cell will link to the protocols in the design section relevant to that relationship.

Lifespan Rate of Protein
Synthesis
PoPS Effect Membrane Traffic
Temperature 1 3
Media 2 4
Parts
Piping

Things to look out for

  • Comparison with the efficiency of other chassis
  • Limitations of type of proteins made in the vesicles
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