User:Vincent Rouilly/Hands On BioBrick Activity

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Vincent 13:42, 6 June 2007 (EDT): feel free to comment on the content of the activity described and its design.

Context

Intentions behind the activity

  • To set the background in terms of Molecular and Celluar Biology:
    • DNA, Central Dogma, Cells and Bacteria
    • Genetic Engineering techniques
  • To touch on Synthetic Biology concepts:
    • BioBricks
    • Modularity
    • Re-usability
  • To give an opportunity to build simple genetic circuits based on opto-electronic components:
    • Protein Generator
    • Inverter, Toggle switch & Repressilator
    • Link circuits with inputs such as: temperature sensor, Infra-red sensor, Touch sensor, Humidity sensor.
    • Link circuits with outputs such as: flagella motor, secretion fan.

Overview

Overview General setting
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  • the activity stands on an oval table, decorated as an E.Coli bacteria.
  • on the right side of the table, DNA, gene expression machinery are illustrated.
  • the hands on activity is placed on the left side:
    • a set of available biobricks implemented as Lego bricks filled with some opto-electronic components.
    • a conductive surface
    • a set of fiber optics to link the different genetic circuits.
  • the activity offers a set of possible inputs for the genetic circuits to be built:
    • temperature sensor (to illustrate heat shock response).
    • infra-red sensor - with remote - (to illustrate UV response)
    • touch sensor
    • humidity sensor
  • 2 outputs are also available:
    • motor driving some flagella
    • fan to illustrate secretion

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Components and Schematics

Constitutive Promoter Activated Promoter Repressed Promoter
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  • Strength of the promoter is set through the potentiometer
  • A battery is embedded into the lego brick
  • The Lego brick is sealed with a conductive plate to make contact with the table.
  • Activity of the promoter is driven by the amount of light received on the photo-sensor.
  • A battery is embedded into the lego brick
  • The Lego brick is sealed with a conductive plate to make contact with the table.
  • Activity of the promoter is driven by the amount of light received on the photo-sensor.
  • A battery is embedded into the lego brick
  • The Lego brick is sealed with a conductive plate to make contact with the table.
RBS Protein Coding Region Stop Codon
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  • Strength of the RBS is set through the potentiometer
  • a 10-LEDs graph bar is used to indicate the level of expression.
  • a LED is used separately to potentially drive another promoter through a fiber optic.
  • the Resistor-Capacitor is used to add a delay.
  • The stop codon enables the circuit to connect to earth through the table

Basic Gene Expression Circuits

Protein Generator Fusion Protein Generator Monocystronic Protein Generator
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  • the protein expression level is tuned by the potentiometers of the promoter and the RBS
  • the protein expression level can be picked up with a fiber optic to drive another promoter or an output modules (flagella motor or secretion fan).
  • the expression level of both proteins is tuned by the potentiometers of the promoter and the RBS. It is the same for both proteins.
  • the protein expression levels can be picked up with a fiber optic to drive another promoter or an output modules (flagella motor or secretion fan).
  • control over the protein expression levels is tuned by the potentiometer of the promoter, but also, independently controlled by the potentiometers of the RBSs.
  • the protein expression levels can be picked up with a fiber optic to drive another promoter or an output modules (flagella motor or secretion fan).

Elementary Genetic Circuits

Inverter Repressillator
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Material

Biobricks

  • constitutive promoter:
    • Duplo brick 2x2
    • potentiometer
    • connector
    • battery
    • electronic circuit
  • ribosome binding site
    • Duplo brick 2x2
      • potentiometer to tune strength of RBS
      • battery
    • connectors x2
  • Stop codon
    • Duplo brick 2x2
    • Connector x1

Inputs Sensors

Motors


Electronic Circuits

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