Difference between revisions of "Team:William and Mary/Basic Part"

 
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Basic Part
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<a href = "http://parts.igem.org/Part:BBa_K2066550"> Bba_K0026550</a> is a decoy binding array containing 85 TetO repeat sequences, and was derived from an addgene plasmid from Finney-Manchester et al. (2013) (“Harnessing mutagenic homologous recombination for targeted mutagenesis in vivo by TaGTEAM). This part can be used to shift the transfer function of an aTC inducible promoter to the left (Figure 1). See decoy binding array page for more information.
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This part was assembled using restriction cloning with EcoRI and SpeI, as accurately performing PCR on the sequence is close to impossible due to the large stretches of homologous sequences.
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<b>Figure 1:</b> Population level FACs data comparing the relative fluorescence of a pTet sfGFP and TetR reporter with and without a tetO binding array. While the data is noisy, it is clear that the inflection point of the circuit with the binding array has shifted to the left as expected. Additionally, both circuits experienced a decrease in fluorescence at higher aTC concentrations, this is likely due to the toxicity of aTC in high concentrations. (See binding array page for more details)
  
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A <b>basic part</b> is a functional unit of DNA that cannot be subdivided into smaller component parts. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0051">BBa_R0051</a> is an example of a basic part, a promoter regulated by lambda cl.
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<p>Most genetically-encoded functions have not yet been converted to BioBrick parts. Thus, there are <b>many</b> opportunities to find new, cool, and important genetically encoded functions, and refine and convert the DNA encoding these functions into BioBrick standard biological parts. </p>
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<h4>Note</h4>
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<p>This page should list all the basic parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page.</p>
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Latest revision as of 21:47, 1 December 2016

<"img src="https://static.igem.org/mediawiki/2016/c/ce/T--William_and_Mary--rubik-background.jpg" width="800px">>

Basic Part

Bba_K0026550 is a decoy binding array containing 85 TetO repeat sequences, and was derived from an addgene plasmid from Finney-Manchester et al. (2013) (“Harnessing mutagenic homologous recombination for targeted mutagenesis in vivo by TaGTEAM). This part can be used to shift the transfer function of an aTC inducible promoter to the left (Figure 1). See decoy binding array page for more information.

This part was assembled using restriction cloning with EcoRI and SpeI, as accurately performing PCR on the sequence is close to impossible due to the large stretches of homologous sequences.

Figure 1: Population level FACs data comparing the relative fluorescence of a pTet sfGFP and TetR reporter with and without a tetO binding array. While the data is noisy, it is clear that the inflection point of the circuit with the binding array has shifted to the left as expected. Additionally, both circuits experienced a decrease in fluorescence at higher aTC concentrations, this is likely due to the toxicity of aTC in high concentrations. (See binding array page for more details)