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

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 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
 * 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.

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)