Team:UC Davis/Project

From 2011.igem.org

Revision as of 01:26, 29 September 2011 by TimFenton (Talk | contribs)

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Start a Family

Got a favorite BioBrick? Check our our process for expanding basic parts into part families.

Criteria

View our judging criteria for iGEM 2011 here.

Overview

We set out to develop a rugged process for the rapid production of mutant libraries of any BioBrick part using standard primers and a simple mutagenic PCR protocol. We chose to prototype this process by creating mutant libraries of the LacI, TetR and λ cI repressible promoters and to mutate GFP to visually assess our ability to create functional protein mutants.

As of October 2011, we have a functioning mutant library generation and screening process, and a selection of well-characterized promoter mutants, including seven LacI promoter mutants and seven λ cI promoter mutants. We also have nine TetR promoter mutants and eight GFP mutants (two of which have been lovingly named "Orange-Mutated Green Fluorescent Protein," or "OMGfp" 1 and 2) which await further characterization.

Project Selection

Why would we want to make mutants? What's so special about repressible promoters? Read about how we came to decide on this project idea here.

Process

Want to make your own mutant library? Curious as to how we assayed our promoter mutants, or how we selected variants that had well-spaced activity levels? Read about our library generation process here.

Promoter Mutants

We chose repressible promoters because they are useful in designing complex circuits and are relatively easy to screen for changes in activity level. You can view detailed information on our LacI, TetR and λ cI mutants on their respective pages.

KO3D Plotting Library

When researching ways to present LacI characterization data clearly on this website, we realized there were no simple, cross-platform javascript libraries for interactive 3D data plotting. To rectify this, we coded our own. Read more about it here.