IGEM:MIT/2005/InsulinProcessor

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Contents

Members

  • Annie
  • Jenny

Goals

  1. Testing bricks in lab to make sure they work
  2. Switching parts
  3. Transfer curve
  4. Quantification

Information

  1. Constitutive promoter for LacI: which strains have it?
 -R0053 from p22 cII 
 -R0040 from TetR
 -Z0251 from T7 
R0040 is stronger than R0053.

Resources

Papers

Summary:
Summary: contains the sequence for LacI, the CAP binding site, lacI repressor binding site. There are variation in the sequence between different strains. A hard copy of the sequence is in the iGEM binder (it's red and kept by Annie).

Books

A Genetic Switch: Phage Lambda Revisited by Mark Ptashne

  • cI and cro are adjacent on the lambda chromosome and transcribed in opposite directions (divergently).
  • cI promoter = Prm points polymerase leftward.
    • Both promoters do not overlap.

cro_rep2.gif L2Fig1_16.jpg L2Fig1_12.jpg

Experiments

  • The production of lacI
-Components: constitutive promoter, RBS + gene sequence + terminator of LacI gene
-Test constructs:
  1. assemble R0053 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
  2. assemble R0040 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
  3. assemble Z0251 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)


  1. assemble R0053 + P0412 (LacI gene)
  2. assemble R0040 + P0412 (LacI gene)
  3. assemble Z0251 + P0412 (LacI gene)
-Measurements:
 Qualitative - 
 Quantitative - 
  • The production of cI/TetR
-Components: CAP binding site, promoter regulated by LacI, RBS + gene sequence + terminator of cI/TetR gene
-Test contructs:
  1. R0010 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
  2. R0010 + P0451 (cI without a cI regulated promoter)
  3. R0010 + Q04510 + E0040/E0433 (Q04510 is cI with a cI regulated promoter)


  1. R0010 + P0440 (TetR without a TetR regulated promoter)
  2. R0010 + Q04400 + E0040/E0433 (Q04400 is TetR with a TetR regulated promoter)

Questions/Suggestions

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