Team:UC Davis/Project

From 2011.igem.org

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<h1>Overview</h1>
<h1>Overview</h1>
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We set out to develop a rugged process for the rapid production of mutant libraries of any <a href="http://biobricks.org/" target:"_blank">BioBrick</a> part using standard primers and a <a href="https://2011.igem.org/Team:UC_Davis/Protocols#ER-PCR">simple mutagenic PCR protocol</a>. 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.<br><br>
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We set out to develop a quick, easy process for the expansion of basic parts into a part families. Our method employs a suped-up <a href="https://2011.igem.org/Team:UC_Davis/Protocols#ER-PCR">mutagenic PCR protocol</a> that uses standard VF2 and VR primers and materials most iGEM teams already have on hand. We chose to prototype this process by creating a part family from the LacI promoter R0010, and to mutate GFP to visually assess our ability to create functional protein mutants.<br><br>
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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.  We also have nine TetR promoter mutants, seven λ cI 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.
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As of November 2011, we have a functioning part family generation process and seven <a href="https://2011.igem.org/Team:UC_Davis/Data_LacI">well-characterized</a> LacI 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.<br><br>
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We have also begun the process of generating part families from the TetR (R0040) and Lambda c1 (R0051) promoters, which are currently being selected for screening.
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<h2>Process</h2>
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<h2>Make a Part Family</h2>
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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 <a href="https://2011.igem.org/Team:UC_Davis/Process">here</a>. <br>
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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 <a href="https://2011.igem.org/Team:UC_Davis/PartFamilies">here</a>. <br>
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Latest revision as of 20:15, 23 October 2011

Our Sponsors

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 quick, easy process for the expansion of basic parts into a part families. Our method employs a suped-up mutagenic PCR protocol that uses standard VF2 and VR primers and materials most iGEM teams already have on hand. We chose to prototype this process by creating a part family from the LacI promoter R0010, and to mutate GFP to visually assess our ability to create functional protein mutants.

As of November 2011, we have a functioning part family generation process and seven well-characterized LacI 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.

We have also begun the process of generating part families from the TetR (R0040) and Lambda c1 (R0051) promoters, which are currently being selected for screening.

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.

Make a Part Family

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 to mutate 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 part families 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.