Team:UC Chile/Cyanolux/Results

From 2012.igem.org

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[[File: LuxABvf_digestion.jpg| 300px| right]]
[[File: LuxABvf_digestion.jpg| 300px| right]]
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<h2>pSB1A3_IntC (From 2010 Utah's iGEM team)</h2>
 
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We decided to use this plasmid to place the LuxCDEG part of the Lux operon in this plasmid, however after various attempts to transform Synechocystis without success we reconsidered (see below SOMEWHERE!!! [[]]) due to problems regarding the design of this plasmid backbone. After analyzing the sequence of the vector (CODE HERE) we blasted the sequences of the recombination sites and we found out that position of the sequences of each recombination site are swapped, leaving the backbone part of the plasmid to be integrated into the genome. Furthermore, the recombination sites are separated by aproximately 2.2 kilobases, knocking 1 gene.
 
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[[File: Construct Recombination sites position.jpg| 500px| center]]
 
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You can see their original construct design [https://2010.igem.org/Design_usu.html#Overview_of_Integration| here].
 
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Here we illustrate the recombination mechanism in which the Int_C plasmid would integrate the backbone part of the vector to the genome.
 
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[[File: Position in genomic DNA.jpg| 300px| right]]
 
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[[File:UC_Chile-CN2Y_Possible_Recombination_Results_and_debugging.jpg|900 px|center]]
 
<h2>pSB1C3_IntS</h2>
<h2>pSB1C3_IntS</h2>
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<h1>Synechocystis</h1>
<h1>Synechocystis</h1>
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<h2>Standardizing growth conditions</h2>
 
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While we managed to acquire an inoculum of Synechocystis PCC 6803, no conditions were set for working with Synechocystis PCC 6803 in our faculty. We proceeded to standardize our own growth conditions by building a custom made cyanobacterial incubator based on growth conditions described in the literature (References?)
 
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Here we show our growth curve for Synechocystis PCC 6803 using the conditions set in our [[Team:UC_Chile2/DIY | DIY]]) shaking incubator (40uE/s/m²), 90 RPM, 30°C.
 
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Methods for the growth curve characterization of Synechocystis PCC 6803 over [[Team:UC_Chile2/Protocols#SyneGrowth | here]].
 
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[[File: Figure for the growth curve.jpg| 500px |center]]
 
<h2>Transforming Synechocystis PCC 6803</h2>
<h2>Transforming Synechocystis PCC 6803</h2>

Revision as of 07:18, 26 September 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012