IGEM:MIT/2005/InsulinProcessor

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

• Autoregulation and Function of a Repressor in Bacteriophage Lambda

Summary:

• E.Coli lactose operon with lacI, lacZ and lacA genes

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.

http://www.clunet.edu/BioDev/omm/repressor/images/cro_rep2.gif http://bioweb.wku.edu/courses/biol566/Images/L2Fig1_16.jpg http://bioweb.wku.edu/courses/biol566/Images/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)