Latest revision as of 03:53, 27 October 2012

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

Project status

Currently, we have been able to detect light-emission from our Synechocystis cells, however we still need to analyze circadian expression of the genes. Furthermore, we still have not been able to assemble the substrate production enzymes to the susceptibility plasmid. We hope that with out new strategy we will be successful. Once we are able to integrate the substrate production genes we may be able to assess the desired behavior of our system and analyze its impact on the chassis.

After having the complete system we will have to see if we can achieve visible lighting with Luxilla. If not, we will try optimizing codon usage for Synechocystis and/or changing promoters.

Biological lighting

Synechocystis PCC6803 has been proposed as the green E. coli. This chassis can be used in industry to achieve more sustainable processes.

Context in Synbio

Since we started developing this project we have realized the importance of biological context, specially when functionality is a desired aspect of the behavioral output of the system. While in one chassis a Biobrick may provide a novel function, in another it may become a practical application for society.

We have seen first hand the difficulties associated with this aspect of current progress in SynBio. By lacking standarized and proven methodologies and relying on previous uncharacterized work. We urge the Synbio community to address this aspect by encouraging the use and characterization of different chassis. It is of utmost importance that BioBrick designed for the chassis must be characterized in the chassis or they may prove fallible. The same can be said about failed experiences with BioBricks that are not reported.

Click here to see our video about sinthetic biology in our lab.

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 {{UC_Chile4}}
-<h1>Promoters Redesigned</h1>
+<h1>Project status</h1>
-We [https://2012.igem.org/Team:UC_Chile/Results/LuxBrick#N-decanal_Effect_Characterization_-_.28BBa_K325909_and_BBa_K325905.29 have seen] that even if induced with substrate, our luciferase constructs don´t show luminescence, though the constructs´ sequences are verified and well characterized luciferases do show light emition.
-Our advisors in cyanobacteria have told us that sometimes promoter sequences can be placed several hundred of bases upstream the +1 site, even inside protein coding sequences.
-That´s why we are designing new versions of our chosen promoters. Now we have changed the promoter construction methodology in terms of the lenght of the upstream sequence considered.
-Instead of considering just 150-200bp, up to 1000bp will be amplified from Synechocystis genome.
+Currently, we have been able to detect light-emission from our <i>Synechocystis</i> cells, however we still need to analyze circadian expression of the genes. Furthermore, we still have not been able to assemble the substrate production enzymes to the susceptibility plasmid. We hope that with out new strategy we will be successful. Once we are able to integrate the substrate production genes we may be able to assess the desired behavior of our system and analyze its impact on the chassis.
-<h1>Transcriptional verification</h1>
+After having the complete system we will have to see if we can achieve visible lighting with <i>Luxilla</i>. If not, we will try optimizing codon usage for Synechocystis and/or changing promoters.
-Even if our promoters are o.k, colony PCRs can´t tell us wether the transcriptional machinery of our cyanobacteria is recognizing our constructs.
-We are designig new primers to make  RT-PCRs that unmistakably verify transcription of the Lux genes  at the specified hours.
+<h1>Biological lighting</h1>
+Synechocystis PCC6803 has been proposed as the green E. coli. This chassis can be used in industry to achieve more sustainable processes.
+<h1>Context in Synbio</h1>
+Since we started developing this project we have realized the importance of biological context, specially when functionality is a desired aspect of the behavioral output of the system. While in one chassis a Biobrick may provide a novel function, in another it may become a practical application for society.
+We have seen first hand the difficulties associated with this aspect of current progress in SynBio. By lacking standarized and proven methodologies and relying on previous uncharacterized work. We urge the Synbio community to address this aspect by encouraging the use and characterization of different chassis. It is of utmost importance that BioBrick designed for the chassis must be characterized in the chassis or they may prove fallible. The same can be said about failed experiences with BioBricks that are not reported.
+[http://biotecnologia.uc.cl/?page_id=2649 Click here to see our video about sinthetic biology in our lab.]
-<h1>Cyrcadian expression characterization</h1>
-We have allready built part [http://partsregistry.org/Part:BBa_K743018 BBa_K743018] to characterize differential expression of sfGFP at different moments of the day.
-Other promoters (new ones or those allready built) will be placed by means of Gibson assembly controlling the expression of this reporter.
-<h1>New Biosafety strategies</h1>
-We have allready assembled part [http://partsregistry.org/Part:BBa_K743010 psb1C3_IntS] wich has a double goal: to insert LuxCDEG genes into Synechocystis and to confere a copper suceptibility that makes this cells unable to thrive in the enviroment.
-We are also designing primers to built our [https://2012.igem.org/File:UC_Chile-Animacion-mE-gen.gif new biosafety system ] based on mE genes and MgSO4, wich will be tested as soon as it is ready.
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Team:UC Chile/Cyanolux/Future

From 2012.igem.org

Latest revision as of 03:53, 27 October 2012

Project status

Biological lighting

Context in Synbio